Nonfermentative (or oxidative) Gram Negative Rods

– Pseudomonas aeruginosa
– Other Pseudomonas species
– Acinetobacter
– Alcaligenes
– Moraxella
– Oligella
– Flavobacterium
– Eikenella
– Kingella
– Stenotrophomonas
– Burkholderia
– others

Nonfermentative Gram Negative Rods

• Don’t form spores
• Cannot break down carbohydrates under anaerobic conditions (TSI=nonreactive)
• Most present in soil, water, plants, and food
• Not fastidious
• Usually grow on BAP and Mac
• Some can grow at room temp
• Resistance to antibiotics (aminoglycosides, cephalosporins, imipenem, penicillin)

Oxidative-fermentative tests (OF)

One tube open to air; one tube overlayed with mineral oil (closed)
oxidizer and fermenter: open yellow; closed yellow
oxidizer and nonfermenter,: open yellow; closed green
nonoxider and nonfermenter: open green; closed green

Oxidase test
• An organism’s oxidase reaction is an important identification clue. NFB may resemble the
Enterobacteriaceae on culture media. The oxidase test sometimes helps to distinguish these two
groups of organisms. Although many nonfermenters are oxidase positive, the Enterobacteriaceae
are oxidase negative. If an organism is oxidase positive, it is NOT an enteric.

Growth on MAC agar

• Many NFB are capable of growing on MAC agar. Those that do produce lactose-negative clear
colorless colonies.

Pseudomonas
• NLF Grown on MAC
• Oxidize glucose
• Reduce nitrate
• Oxidase positive
• Contain P. aeruginosa (only pyocyanin producing NF)
– Fluorescent group: P. putida and P. fluorescens (and P. aeruginosa)
– Stutzeri group: P. stutzeri, P. mendocina, and others

Pseudomonas aeruginosa
• Infections:
– Severe wound infections in burn patients
– Chronic lung infections in patient with cystic fibrosis
– Swimmer’s ear—hot tub syndrome

• Colony characteristics:
– Large, irregularly shaped, and have a blue green metallic sheen
• Pyocyanin (blue green pigment)
– Beta-hemolytic on BAP
– Grapelike odor

• Key identification tests:

– Oxidase positive
– TSI: K/K
– Pyocyanin production and Grows at 42 C
• Hard to treat—resistant to many antibiotics
• Conventional biochemical testing—kits
• Few biochemical tests
– Oxidase (mostly positive)
– Growth on MacConkey agar
– Glucose utilization (can’t ferment)
– Motile

Burkholderia
• B cepaci: (once known as Pseudomonas cepacia)
– Important nosocomial pathogen
• Severe respiratory disease in patients with CF
• Grows on routine lab media, including MAC; strong earthy odor, may have yellow pigment
• Weakly oxidase positive, oxidizes many carbohydrates, motile

Stenotrophomonas maltophilia
• Uibquitous, common to water, sewage, and plant material
• Common in hospital—contaminant in blood-drawing equipment, disinfectants, etc.-
Nosocomial
– Immunocompromised
• Resistant to many antibiotics
• NEGATIVE OXIDASE REACTION
• ammonia like odor
• Grows on MAC, is oxidase negative and oxidizes glucose

Acinetobacter
• Acinetobacter baumannii and Acinetobacter lwoffii
• Ubiquitous in environment and hospital (ventilators, humidifiers, catheters)
• Opportunistics
• OXIDASE NEGATIVE, NONMOTILE