non-fermenting and miscellaneous gram negative bacilli
TRANSCRIPT
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NON-FERMENTING AND MISCELLANEOUS GRAM NEGATIVE BACILLI
GROUP 6, 3GMTPeralta, Ana Eryka Elaine
Pilares, Felicia AnnRafols, Kate Syvel
Sales, Reneepearl KimSato, Mikie
Sison, Jacqueline Anne
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ClassificationPseudomonads
Pseudomonas aeruginosa (most common)Pseudomonas stutzeriBurkholderia pseudomalleiBurkholderia cepaciaStenotrophomonas maltophilia (also common)
Non-PseudomonadsAcinetobacter baumannii MoraxellaAchromobacter xylosoxidans
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General Characteristics• Obligate aerobes• All are motile (except Burkholderia mallei)• Non-fermentative for glucose• TSI reaction: K/K
• Oxidative for glucose (except Moraxella – asaccharolytic)• Hugh-Leifson O-F glucose positive
• Cytochrome oxidase positive (except Acinetobacter and
S. maltophilia)
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General Characteristics
Cytochrome oxidase
OF glucose Motility Pyoverdin* production
Susceptibility to Polymixin B
P. aeruginosa**
+ + + +(also P.
fluorescens and P. putida)
+
P. stutzeri + + + - +
Burkholderia cepacia
+ + + -
Burkholderia pseudomallei
+ + + -
Stenotrophomas maltophilia
- +(also OF
maltose ++)
+ +
Pseudomonads
*Pyoverdin is a fluorescent, yellow-green or yellow-brown water-soluble pigment**Of the Fluorescent group, P. aeruginosa solely produces Pyocyanin, a blue water soluble pigment;
P. fluorescens and P. putida cannot
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General CharacteristicsNon-Pseudomonads
Cytochrome oxidase
OF glucose Motility
Acinetobacter baumanii
- +(also OF
lactose ++)
-
Moraxella + -(asaccharolytic*)
+/-
Achromobacter xylosoxidans
+ +(also OFXylose +)
+
*Assacharolytic means that it alkalinizes (rather than acidifies) the broth in Hugh-Leifson tube unsealed with mineral oil
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Colony Characteristics
Colony Characteristics
Pseudomonas aeruginosa
Spreading and flat with a pearl-like sheen (pearlescent) & grape-like odorGrow at 42˚C
Pseudomonas stutzeri
Produce distinctive dry, wrinkled colonies (1-6mm) on BAP
Burkholderia cepacia
Smooth, round, opaque, & yellow colonies on BAPBright pink colonies on MAC (due to lactose oxidation)
Stenotrophomonas maltophilia
Rough, lavender-green colonies with ammonia odor on Sheep Blood Agar
Pseudomonads
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Colony Characteristics
Colony Characteristics
Acinetobacter spp.
Smooth, opaque, gray-white colonies (slightly smaller than Enterobacteriaceae)
A. baumanii – non-hemolyticA. iwoffii – non-hemolyticA. haemolyticus – beta hemolytic on Sheep Blood Agar
Moraxella Small (0.5-1mm), smooth, translucent to semi-opaque colonies on BAP
Non-Pseudomonads
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Microscopic Appearance
Microscopic Appearance
Pseudomonas aeruginosa
Straight or slightly curved, slender, Gram (-) rods
Burkholderia cepacia Gram (-) rods
Stenotrophomonas maltophilia
Short to medium sized, straight, Gram (-) rods
Acinetobacter spp. Gram (-) coccobacillary rods occuring singly and in Neisseria-like pairs
Moraxella Gram (-) or Gram-variable Neisseria-like diplococci
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Virulence factors: Fluorescent: Pseudomonas aeruginosa
• Endotoxin from cell wall (lipopolysaccharide)• Motility (Flagella)• Pilli(Type 4)• Neuraminidase- facilitate binding pilli• Capsule• Exotoxins: protease (LasBelastase, LasAelastase, and alkaline protease),
hemolysins (Phospholipase C and Rhamnolipid), lecithinase, elastase, DNAse, exotoxin A, exoenzyme S
• Alginate- polysaccharide polymer, produces mucoid colonies• Resistant to aminoglycosides, ъ-lactam antibiotics, fluoroquinolones• Pyocyanin• Pyoverdin
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Fluorescent: Pseudomonas fluorescens and Pseudomonas putida
• Very low virulence• Resistant to cabernicillin
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Nonfluorescent group:• P. stutzeri• Resistant to Chloramphenicol and Cephalosporin
• P. mendocina• Flagellum, oxidise glucose and xylose, positive for oxidase and
ADH, nonproteolytic• P. pseudoalcaligenes, P. alcaligenes• Oxidase positive
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Stenotrophomonas maltophilia
• Resistant to Cephalosporins, Penicillins, Carbapenems, Aminoglycoside• Positive for:• DNase• Esculin• Gelatin Hydrolysis• Lysine decarboxylase
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Acinetobacter (A. baumanii& A. lwoffii)• S-form LPS- major stimulator of immune
response; resistance to the natural bactericidal action of human serum• Adhesins- formation of biofilm• Lipase(s)- damage tissue lipids• Elaboration of siderophores- to
scavenge iron and promote survival in vivo
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Burkholderia
• Burkholderiacepacia:• Flagella• Lysine decarboxylase positive• o-nitrophenyl-β-d-galactopyranoside (ONPG)
positive• Burkholderia gladioli• one or two polar flagella• Catalase and urease positive• Resistant to polymyxin B
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Burkholderia
• Burkholderiamallei• Burkholderiapseudomallei:• Secretion (type III protein secretion system)• Lipopolysaccharide• Capsule• Exoproteins (Phospholipase C, putative
collogenase, MucD Ser protease homologs)• Adhesins
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Flavobacterium and Chryseobacterium
• Flavobacterium• Chryseobacterium• DNase• Fibrinolysin• Gelatinase• Lecithinase• Lipase• Proteinase
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CultivationP. AREUGINOSA
• Carbon Source- ACETATE• Nitrogen Source – AMMONIUM SULFATE• 37 degrees, able to grow at temperatures as high as 42 degrees• resistant to high concentrations of salts and dyes, weak antiseptics,
and common antibiotics• 1st - large, smooth, with flat edges, elevated• 2nd - a mucoid appearance, attributed to the production of alginate
slime
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P. STUTZERI• Freshly isolated colonies - adherent, wrinkled appearance,
reddish brown• Shapes of colonies are neither uniform nor necessarily
constant: they change appearance with time• After repeated transfers: may become smooth, butyraceous,
pale in color (colonial dissociation)• 30°C and stored at 4°C for 24 hours
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B. PSEUDOMALLEI• Ashdown’s medium• Suppressors: gentamicin, crystal violet• 96 hours, flat wrinkled, purple colonies• Enriched with 4% glycerol• The cost of each plate is only US$0.04 = Php. 1.75
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STENOTROPHOMONAS MALTOPHILIA• smooth, glistening, with entire margins and are white to pale
yellow• on blood agar there may be a greenish discoloration of the
medium around confluent growth• on clear media, some strains may exhibit a brownish
discoloration• optimal at 35°C• methionine or cystine is required for growth by most but not
all strains
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ACOLIGENES XYLOSOXIDANS• flat, spreading and rough colonies• blood agar and MAC agar
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Biochemical Identification of Species
• All the nonfermenters will not yield acidic reactions in the anaerobic portion of media such as TSI (Triple Sugar Iron) or KIA (Kligler’s Iron Agar)
• Thin, gram-negative bacilli or coccobacilli on gram stain may indicate the presence of nonfermenter.
• For clinically significant isolates of P.aeruginosa, S. malthophilia, Acinetobacter spp., and Burkholderia cepacia, the CLSI recommends broth dilution or Kirby Bauer disk diffusion assay to be performed.
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Pseudomonas aeruginosa; Pseudomonas stutzeri;
Burkholderia cepacia; Burkholderia pseudomallei
•TSI (K/K) •Positive in MacConkey agar•Positive in Oxidase test
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P.Aeruginosa• Unipolar motility
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Moraxella atlantae; Moraxella osloensis
•TSI (K/K)•Positive in MacConkey agar.•Positive in Oxidase test
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Acinetobacter baumanii
•TSI (K/K) •Positive in MacConkey agar•Negative in Oxidase test•Usually nitrate negative•Non motile
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Stenotrophomonas maltophilia
•TSI (K/K)•Positive in MacConkey•Negative in Oxidase test
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Clinical Infections and DiseasesNon fermenters account for about 15% of all gram (-) bacilli
isolated clinical specimens
Non Fermenters can be responsible for infection: o Septicemia o Meningitidiso Osteomyelitis o Wound infections (develops after trauma or surgery)
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Risk Factors for Diseases cause by Nonfermentative Gram- Negative Bacilli
ImmunosuppressionTraumaForeign body implatationInfused fluids
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Diseases Produced
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(Jawetz, Melnick & Adelberg,2007)Mahon, Lehman & Manyselis,2010)
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(Jawetz, Melnick & Adelberg,2007)Mahon, Lehman & Manyselis,2010)
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(Jawetz, Melnick & Adelberg,2007)Mahon, Lehman & Manyselis,2010)