CLINICAL

BACTERIOL
OGY
MLSBIO104
NON-
FERMENTATIV
E GRAM
NEGATIVE
BACILLI
OUTLINE:

• General Characteristics
• Epidemiology
• Pathogenesis and Spectrum of Diseases
• Specific Organisms
• Laboratory Diagnosis
• Antimicrobial Susceptibility Testing and
Therapy
• Prevention
 • Identify non-fermentative gram
negative bacilli
• List the most common non-
fermentative gram-negative organisms
• Explain where Acinetobacter spp. are
found and the patients most at risk of
infection.

OBJECTIVE: • Describe the Gram stain morphology of

Acinetobacter, Bordetella, and
Stenotrophomonas spp.
• Describe the appearance and odor of
Stenotrophomonas maltophilia when
grown on blood agar.
• Differentiate between the two groups
of Acinetobacter organisms and
identify the most dependable test to
distinguish between the groups.
GENERAL
CHARACTERISTICS

• nonfermenting
• gram-negative bacilli
• multidrug resistant
• oxidase negative
• generally grow well on MacConkey
(except for the CDC group NO-1)
Epidemiology

• often inhabit environmental niches

• humid conditions are more
favorable for growth.
• capable of survival on inanimate
objects for extended periods
• Both Acinetobacter spp. and S.
maltophilia can contaminate
potable or chlorinated water.
• third and fourth most common
gram-negative bacilli, respectively,
encountered in clinical specimens.
Pathogenesis and Spectrum of
Disease
• Opportunistic pathogens.

• Acinetobacter spp.
• Produce a lipopolysaccharide capsule
• adherence to mucosal epithelium
• involve the respiratory or genitourinary tract, bacteremia, and,
occasionally, wound infections
• S. maltophilia
• high mortality rates in debilitated patients and those who are
immunocompromised.
• involve the respiratory or genitourinary tract, bacteremia, and,
occasionally, wound infections

• Acinetobacter baumannii is typically the species identified among nosocomial

infections.
• Pseudomonas oryzihabitans and CDC group NO-1 are not routinely isolated in
clinical specimens
• Mostly been isolated from wounds, but also occasionally blood and
dialysis fluid cultures.
• Pseudomonas luteola is typically reported in skin infections, endocarditis,
osteomyelitis, and meningitis after neurosurgical procedures.
• Bordetella holmesii is rare, but can cause severe systemic illness as well as
pertussis-like symptoms
Laboratory Diagnosis
Gram stain:
• Acinetobacter spp. are plump coccobacilli
• The Bordetella spp. are coccobacilli or short rods.
• S. maltophilia, P. oryzihabitans, and P. luteola are short to medium-size straight rods.
• CDC group NO-1 are coccoid to medium-size bacilli.

Cultivation:
• TSA, 5% SBAP, CAP, MAC
• Aerobic and anaerobic conditions
 Approach to identification

• Automated identification systems can usually identify the organisms to the species level
• additional testing may be required to speciate the organisms using conventional
biochemical and physiologic characteristics
• Sequence-based methods, including amplification of the ribosomal RNA (rRNA)
sequence, genomic fingerprinting, and restriction endonuclease analysis, have been used
to identify Acinetobacter spp.
• Acinetobacter has 34 named species that have been placed into homologous groups
(genomospecies) based on deoxyribonucleic acid (DNA)-DNA hybridization studies.
• Acinetobacter species are oxidase negative, catalase positive, and nonmotile.
• Acinetobacter baumannii (glucose-oxidizing, nonhemolytic strains)
• Acinetobacter lwoffi (non–glucose utilizing, nonhemolytic strains)
• Acinetobacter haemolyticus (Beta-hemolytic strains)
• S. maltophilia is an oxidase-negative, nonfermentative, gram-negative bacilli
• brown pigment on brain-heart infusion agar that contains tyrosine.
• Pseudomonas spp. are gram-negative, nonfermentative, oxidase-negative, catalase-
positive bacilli.
• CDC group NO-1 bacteria are oxidase negative, asaccharolytic, and nonmotile and
typically form small colonies on blood agar.
 Antimicrobial
Susceptibility
Testing and Therapy
• intrinsic mechanisms, which impart broad-spectrum antibiotic resistance
• Acinetobacter spp. may be multidrug resistant (MDR) or extremely drug resistant
(XDR) (AmpC betalactamases)
• acquire beta-lactamase enzymes that reside in mobile genetic elements
• S. maltophilia exhibits resistance to a wide-range of antibiotics including
betalactams, cephalosporins, aminoglycosides, tetracyclines, and polymyxins
through reduced membrane permeability, various enzymes, or efflux pumps.
Acinetobacter spp.
• Typical first-line agents include broad-spectrum cephalosporin, a
beta-lactam/beta-lactamase inhibitor combination agent, or a carbapenem.
(Ertapenem should not be used)
• For drug-resistant infections, polymyxins (e.g., colistin), minocycline, or
tigecycline
S. maltophilia
• Trimethoprim-sulfamethoxazole - recommended as the primary drug of choice.
• ticarcillin-clavulanic acid, fluoroquinolones, tigecycline, minocycline, rifampin,
or moxifloxacin should be considered if the organism tests susceptible
Prevention

• health care settings and patient environments are

cleaned regularly to prevent environmental
contamination.
• Care should be taken to follow the proper aseptic
techniques for the insertion of central venous
catheters and other indwelling medical devices.
• All reusable medical equipment should be
properly decontaminated or sterilized.
END