Chapter 21

Nonfermenting and Miscellaneous

Gram-Negative Bacilli

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 General Characteristics of
Nonfermenters
 Do not ferment sugars (oxidative-fermentative [O/F]
media)
 Oxidize (saccharolytic) sugars
 Their ability to use a variety of sugars is diagnostic
 Nonoxidizers (asaccharolytic)
 Do not break down carbohydrates at all and are inert or
biochemically inactive
 Long, thin, gram-negative bacilli or coccobacilli
 Generally oxidase positive (occasionally negative)
 Triple sugar iron (TSI) butts do not acidify
(nonfermenter)

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 Clinical Infections
 15% of all gram-negative bacilli isolated
 Septicemia
 Meningitis
 Osteomyelitis
 Wound infections

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Risk Factors for Disease

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Grouping of Nonfermenters via TSI,
MacConkey (MAC), Oxidase, and
Oxidation

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Characteristics Common to Groups
of Nonfermenters

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Taxonomic Changes for Some
Gram-Negative Nonfermenters

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Biochemical and Morphologic
Characteristics of Select
Nonfermenters

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 Biochemical and Morphologic
Characteristics of Select
Nonfermenters (Cont.)
 Definitive identification
 Usually reference lab using nucleic acid
sequencing or mass spectrophotometric methods
 Matrix-associated laser desorption-ionization time-
of-flight (MALDI-TOF)
• Identification of bacteria and fungi from colonies that is
based on analysis of the unique spectra of their peptides
when separated in the mass spectrophotometer
 Mean time for identification is 6 min

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Pseudomonas Fluorescent Group
 P. aeruginosa
 Most commonly isolated species
• Leading cause of nosocomial respiratory tract infections
 Grapelike odor, blue-green pigment
 Oxidase and catalase positive
 Oxidizes carbohydrates

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 P. aeruginosa
 Clinical infections
 Wound infections and burn patients
 Pulmonary infections (cystic fibrosis [CF] patients)
 Nosocomial urinary tract infections (UTIs) and
bacteremias
 Endocarditis
 Ear infections
 Skin rashes
• Hot tub syndrome

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 P. aeruginosa (Cont.)
 Virulence factors
 Endotoxins
 Exotoxins
• Particularly exotoxin A
 Motility
 Pili
 Proteases
 Hemolysin
 Capsule
 Lecithinase
 Elastase
 DNase
 Alginate
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 P. aeruginosa (Cont.)
 Identification characteristics
 Green sheen on blood agar
• Specific to P. aeruginosa
 Pyoverdin
– Yellow-green or yellow-brown pigment
 Pyocyanin
– Blue pigment
 Most are β-hemolytic
 Arginine dihydrolase (ADH) positive
 Growth at 42°C
 Citrate positive
 Cetrimide agar
• Selective and differential media
 Very drug resistant, but aminoglycosides work best
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Gram Stain and Colonies of P.
aeruginosa

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 P. fluorescens and P. putida
 Fluorescent group
 Low virulence
 Isolated from respiratory specimens, contaminated blood
products, urine, cosmetics, hospital equipment, and fluids
 Infections
 UTIs
 Postsurgical abscesses
 Empyema
 Septic arthritis
 Wound infections

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P. fluorescens and P. putida (Cont.)
 Identification characteristics
 P. fluorescens and P. putida produce pyoverdin, but neither
produces pyocyanin.
 Does not grow at 42°C
 Cannot reduce nitrate to nitrogen gas
 Can produce acid from xylose
 Gelatin hydrolysis
• P. putida is negative.
• P. fluorescens is positive.
 Treatment
 Susceptible to aminoglycosides, polymyxin, and piperacillin
 Resistant to carbenicillin and trimethoprim/sulfamethoxazole (SMZ)

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 Nonfluorescent Group
 P. stutzeri
 Easily recognizable
• Wrinkled, leathery, adherent colonies that may produce a light
yellow or brown pigment
 Identification characteristics
 Arginine dihydrolase negative
 Starch hydrolysis positive

 Treatment
 Susceptible to the aminoglycosides, SXT, ampicillin polymyxin,
tetracyclines, fluoroquinolones, and third-generation
cephalosporins
 Resistant to chloramphenicol and the first- and second-
generation cephalosporins

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 P. stutzeri

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 Nonfluorescent Group (Cont.)
 P. mendocina
 Contaminant
 P. pseudoalcaligenes
 Arginine dihydrolase positive and will weakly
ferment fructose
 Contaminant

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 Nonfluorescent Group (Cont.)
 P. alcaligenes
 Contaminant
 P. luteola and P. oryzihabitans
 Rare
 P. luteola can be differentiated from P.
oryzihabitans by ortho-Nitrophenyl-β-galactoside
(ONPG) and esculin hydrolysis.

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 Acinetobacter Species
 Second most common isolate of these organisms
 Sometimes a normal flora: 25% of adults
 45% of tracheostomy sites
 Found in hospitals particularly ventilators, humidifiers,
catheters
 UTIs, pneumonia, endocarditis, septicemia,
meningitis, wounds, burns, eye infections
 A. baumannii
• Glucose-oxidizing, nonhemolytic strain
 A. lwoffii
• Glucose-negative, nonhemolytic strain

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Photographs of A. baumannii

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 Stenotrophomonas maltophilia
 Third most common isolate of these
organisms
 Commonly contaminate blood drawing equipment,
disinfectants, transducers, and other equipment
 Pneumonia, endocarditis (surgery and
intravenous [IV]-drug users), bacteremia
 Rarely meningitis and UTIs

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 S. maltophilia (Cont.)
 Identification characteristics
 Catalase positive, oxidase negative
 DNase positive, esculin positive, lysine
decarboxylase positive
 Gelatin hydrolysis positive
 Usually susceptible to SXT

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 Burkholderia cepacia
 Nosocomial pathogen
 Pneumonia particularly in CF patients
 Endocarditis, UTIs, osteomyelitis, dermatitis,
wound infections

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 B. cepacia (Cont.)
 Identification characteristics
 Oxidase negative, motility positive
 Oxidizes glucose, maltose, lactose, mannitol
 Lysine decarboxylase (LDC) positive
 Ornithine decarboxylase (ODC) positive
 Arginine dihydrolase (ADH) negative

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B. cepacia on Selective Media

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 Other Burkholderia
 B. gladioli
 Resembles B. cepacia
• Can be found in CF and chronic granulomatous disease
(CGD) patients
 B. mallei
 Causes glanders
• Can cause severe suppurative or acute pulmonary
infections
• One case in 50 years (laboratory accident)

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 Other Burkholderia (Cont.)
 B. pseudomallei
 Melioidosis
• An aggressive granulomatous pulmonary disease
 May cause overwhelming septicemia

 Transmission
 Ingestion, inhalation, or inoculation
 Possible agent of bioterrorism

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B. pseudomallei on Sheep Blood

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 Moraxella, Oligella, and
Psychrobacter
 Normal flora of the mucous membranes
 Negative for carbohydrate oxidation
 Strict aerobes
 Moraxella
 Oxidase positive; nonmotile, gram-negative,
coccobacilli
• Isolated from respiratory tract, urinary tract, and the eye
 Rarely cause disease, except M. catarrhalis
 M. nonliquefaciens is part of the normal flora.

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 Moraxella, Oligella, and
Psychrobacter (Cont.)
 Oligella
 Small, paired, gram-negative bacilli or coccoid
organisms
• Most often isolated from the urinary tract
 Potato dextrose agar (PDA) positive, oxidase
positive, nitrate and nitrite positive with gas, and
motile
 Psychrobacter immobilis
 Nonmotile, oxidase positive, oxidative diplococcus
 Optimal temperature for growth is 20°C.

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 Alcaligenes and Achromobacter
 Isolated from hospitalized patients
 Urine, stool, sputum, wounds, blood
 Identification characteristics
 No oxidation in O/F media with blue color at the top
• Except Alcaligenes xylosoxidans
 Acid in glucose and xylose
 Motile
 Reduce nitrates to nitrites
• Alcaligenes faecalis (further reduce to nitrogen gas)
 Negative for indole
 Negative for esculin and gelatin hydrolysis

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Flagella Stains of Gram-Negative
Bacilli

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Colonies of Achromobacter

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 Less Commonly Encountered
Nonfermentative Gram-Negative
Bacilli
 Brevundimonas
 Oxidize glucose and are oxidase positive, most
grow on MAC agar
• Esculin hydrolysis to separate species
 Centers for Disease Control and Prevention
(CDC) groups eugonic oxider (EO)-2, EO-3,
EO-4, and Paracoccus.

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 Less Commonly Encountered
Nonfermentative Gram-Negative
Bacilli (Cont.)
 Chromobacterium violaceum
 Wound contamination with soil and water
 Comamonas and Delftia
 Straight to slightly curved motile rods that produce
alkalinity in O/F media
 Catalase and oxidase positive
 Reduce nitrate to nitrite

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 Flavobacteriaceae
 Major revisions to genus
 Balneatrix
 Bergeyella
 Chryseobacterium
 Elizabethkingae
 Empedobacter
 Myroides
 Weeksella
 Wautersiella
 Sphingobacterium
 Several unnamed CDC groups

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 Flavobacteriaceae (Cont.)
 Long, thin bacilli with bulbous ends
 Contaminant of hospital equipment
 Identification characteristics
 Nonmotile, DNase positive, oxidase positive
 Gelatin hydrolysis positive, indole positive
 Elizabethkingia meningoseptica
 Meningitis and septicemia of the newborn (especially
neonates)
 Contaminant of povidone iodine pads
• U.S. Food and Drug Administration (FDA) recall in 2011
 Pneumonia, endocarditis, bacteremia, meningitis

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Growth of E. meningosepticum

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 Less Commonly Encountered
Nonfermentative Gram-Negative
Bacilli (Cont.)
 Balneatrix alpaca
 Pneumonia and meningitis from hot spring spa
 Motility and nitrate reduction should help to
differentiate from Elizabethkingia
 Chryseobacterium indologenes
 Sometimes nosocomial or contaminant
 Empedobacter
 Endophthalmitis from cataract surgery
• Poor sterilization procedure

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 Less Commonly Encountered
Nonfermentative Gram-Negative
Bacilli (Cont.)
 Wautersiella
 Blood and surgical wounds
 Myroides odoratimimus
 Environmental organism
• Can cause UTI, endocarditis, ventriculitis, and cutaneous
infections in immunocompromised patients

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 Less Commonly Encountered
Nonfermentative Gram-Negative
Bacilli (Cont.)
 Weeksella
 Species can be found in genitourinary specimens
and as a cause of cellulitis, tenosynovitis,
septicemia, pneumonia, and meningitis.
• Associated with dog and cat bite wounds
 Sphingobacterium
 Septicemia meningitis, peritonitis, and possible
cellulitis
• Isolated from blood and urine

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 Less Commonly Encountered
Nonfermentative Gram-Negative
Bacilli (Cont.)
 Methylobacterium
 Bacteremia, peritonitis, synovitis, and skin ulcers
• Generally in immunocompromised
 Contaminated tap water has been implicated.
 Roseomonas mucosa
 Immunocompromised, catheter-related blood
infections

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Roseomonas spp.

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 Less Commonly Encountered
Nonfermentative Gram-Negative
Bacilli (Cont.)
 Ralstonia pickettii
 Contaminants of sterile products
• Can be isolated from urine, nasopharynx, abscesses,
wounds, and blood, usually as colonizers or
contaminants
 Cupriavidus paucula
 Opportunistic pathogen that can cause serious
infections
• Septicemia, peritonitis, abscesses, and tenosynovitis
 Generally in immunocompromised patients

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 Less Commonly Encountered
Nonfermentative Gram-Negative
Bacilli (Cont.)
 Shewanella putrefaciens and S. algae
 Infrequent isolates and rarely pathogenic
 Abscesses and traumatic ulcers, otitis media,
ocular infections, osteomyelitis, peritonitis, and
septicemia
• Represent colonization rather than infection

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 Less Commonly Encountered
Nonfermentative Gram-Negative
Bacilli (Cont.)
 Sphingomonas paucimobilis
 Sphingomonas. parapaucimobilis
 Found in water sources including swimming pools,
hospital equipment, and laboratory supplies
• Peritonitis associated with CAPD, septicemia, meningitis,
leg ulcer, empyema, and splenic and brain abscesses
 Additionally isolated in sputum, urine, and vaginal
specimens

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Colonies of Sphingomonas

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