Gram Negative Bacilli/Coccobacilli

(Aerobic)
• MacConkey Agar Growth
• Oxidase (-): Enterobacteriaceae, Acinetobacter, Stenotrophomonas,
• Oxidase (+): Pseudomonas, Burkholderia, Achromobacter, Rhizobium,
Ochrobactrum, Chyseobacterium, Sphingobacterium, Alcaligenes, Bordetella
(non pertussis), Comamonas, Vibrio, Aeromonas, Plesiomonas,
Chromobacterium
• McConkey Agar No Growth
• Growth Require Special Media
Acinetobacter
• Aerobic, Gram negative bacteria
• A. baumannii, A. lwoffii
• Coccobacillary, diplococcal or coccal
• Mimics other etiologic agents if specimen is genital discharge or CSF
• Often are commensals, occasionally nosocomial
• Multidrug resistant
Family Pseudomonaceae
• Pseudomonas aeruginosa, fluorescens, putida, stutzeri
• Burkholderia cepacia, mallei, pseudomallei
• Stenotrophomonas (Xanthomonas) maltophilia
• Comamonas testosteronii, Shewanella putreficans, Brevundimonas
diminuta, Ralstonia pickettii, Ralstonia mannitolliytica
Stenotrophomonas maltophilia
• Lavender green on BAP; yellow tan on TSA
• Direct contact with hospital devices
• Nosocomial infections
Family Pseudomonaceae
• Unusual fruity, grapelike or corn tortilla odor
• Pigments
• All are motile except Burkholderia mallei
• All are non fermentative but oxidative
• All are oxidase (+) except S. maltophilia but ONPG (+)
• Aminoglycosides – tobramycin, gentamicin
• Carbenicillin
Pseudomonas
• Oxidase (+), motile, grape/fruity odor, pigments
• Slime layer, pili, “O” Ag, endotoxin, phosphorylase (destroys pulmonary surfactant)
• P. aeruginosa –“blue/green pus”,“swimmer’s rash”
nosocomial infection, drug resistance, burns
• P. mallei – horses – “Glanders/Farcy’s disease”
• P. pseudomallei – meliodosis (lung infection) – now under Burkholderia
• P. maltophila – oxidase negative
• Opportunistic – P. stutzeri (brown pigment), P.fluorescens, P. putida
Opportunistic pathogens
Pigment Oxidase ONPG

Xanthomonas Yellow (-) (+)

Comamonas None (+) (-)

Shewanella Pink - brown (+) (+)

Pseudomonas
pathogenic determinants
• Exotoxin A – inhibits protein synthesis
• Endotoxin – causes fever, DIC, necrosis
• Pili – causes adhesion to tissue cells
• Alginate – inhibits chemotaxis; phagocytosis
• Intrinsic resistance - unusual porin channels
• Elastase
• Collagenase
• Protease
Pseudomonas aeruginosa
• Dull gray hemolytic with feathery edge; pigments on clear agar
• Ingestion, exposure to contaminated medical devices, skin
penetration
• UTI, skin, wound, burns, nosocomial, swimmer’s ear, Shanghai fever,
jacuzzi or hot tub syndrome, resp tract infection
Lab tests for P. aeruginosa:
• Oxidase (+); K/K on TSI
• Optimum growth at 42 degrees C
• blue green diffusible pigment w/c may turn brown on NA, TSA, MHA
• nitrate reduction
• ADH (+); LDC, LDAm (-)
• Glucose oxidizer, xylose oxidizer but does not ferment any CHO (non
fermentative)
Green Fluorescence under u.v. light
• P. aeruginosa
• P. putida
• P. fluorescens
Pseudomonas fluorescens
• Green pigment on clear agar
• Direct contact with contaminated food, plants
• Food spoilage, UTI, wound and skin infections
Pseudomonas putida
• Green pigment on clear agar
• Skin penetration
• Septicemia, nosocomial infection
Burkholderia stutzeri
• Dry, wrinkled white colonies
• Skin penetration
• Otitis media, conjunctivitis, septic arthritis
Burkholderia cepacia
• Yellow serrated colonies
• Skin penetration
• Nosocomial infection like UTI, wound infection and septicemia; onion
bult rot; foot rot
Burkholderia mallei
• Smooth cream to white colonies
• Contact with infected horse
• Glanders or farcy
MAC + Nonfermentative but oxidative
gram negative bacilli
• MAC: colorless, NLF
• Motile with peritrichous flagella, obligate aerobe, non fermentative
but oxidative
• Opportunistic, nosocomial
• Achromobacter denitrificans
• Nitrate reduction positive; in those w/ cystic fibrosis, small convex glistening
colonies
• Alcaligenes faecalis
• nitrate reduction negative; alpha hemolytic w/ feathery edge;
apple/strawberry-like odor
• Chryseobacterium meningosepticum
Gram Negative Bacilli/Coccobacilli
(Aerobic)
• McConkey Agar Growth
• Oxidase (-): Enterobacteriaceae, Acinetobacter, Stenotrophomonas
• Oxidase (+): Pseudomonas, Burkholderia, Achromobacter, Rhizobium,
Ochrobactrum, Chyseobacterium, Sphingobacterium, Alcaligenes, Bordetella
(non pertussis), Comamonas, Vibrio, Aeromonas, Plesiomonas,
Chromobacterium
• McConkey Agar No Growth
• Growth Require Special Media
MAC (+), oxidase (+),
glucose fermenters,
aerobic / facultative anaerobic
• Family Vibrionaceae
• Vibrio cholerae, parahemolyticus, vulnificus, alginolyticus
• Plesiomonas shigelloides
• Family Aeromonadaceae
• Aeromonas caviae
• Aeromonas hydrophila
Vibrio

• Limulus test – detects endotoxins – visible gel in lysates of washed

amebecytes of horse crab
• Ileal loop test – detects enterotoxins
• Kamagawa phenomenon – V. parahemolyticus exhibits double zone
hemolysis Wagatsuma agar
• Cholera red test – V. cholera is positive in indole and nitrate reduction
tests
Biotypes
El Tor Classical
Hemagglutination + -
using chicken RBC
Hemolyis using + -
sheep RBC
Voges Proskauer + -
Phage IV suscep. - +
Polymxyin B - +
susceptibility
Vibrio/Plesiomonas/Aeromonas
Colonial TCBS – sucrose (-) TCBS (-) TCBS
morphology fermenter – yellow Non fermenter Non fermenter
Non sucrose fermen
- blu/green

Microscopic G(-) curved, comma G(-) comma or G(-) straight bacilli

morphology shaped or straight straight bacilli,
bacilli singly, pairs, short
chains

motility + darting motility + +

Vibrio/Plesiomonas/Aeromonas
habitat Brackish or salt Fresh water esp. in Fresh, brackish
water warmer climate marine, polluted or
chlorinated water

Oxidase test All +) except V. (+) (+)

metschnikovii

Halophilic (grows All (+) except V. No No

on 6 % NaCl) cholerae & mimicus

Growth at Will grow Will not grow Will not grow

pH 8.4 – 9.5

Inositol Non fermenter fermenter Non fermenter

fermentation
Vibrio/Plesiomonas/Aeromonas
Susceptibility to susceptible susceptible resistant
vibriostatic agent
0/129

String test w/ 0.5 % positive negative negative

Na deoxycholate
Vibrio cholerae
• Cholera toxin (choleragen); Zot toxin; Ace toxin; 01
and 0139 somatic antigens; Hemolysin; Cytotoxin;
Endotoxin; Mucinase; pili
• Ingestion
• Cholera – rice watery stools, dehydration –
washerwoman’s hands, sunken eyes, hypotension,
death
• Ogawa, Inaba, Hikojima
CAMP mediated
• The lost H2O and electrolytes in mucosal cells are replaced from the
blood. Thus, the toxin-damaged cells become pumps for water and
electrolytes causing the diarrhea, loss of electrolytes, and
dehydration that are characteristic of cholera.
• cAMP blocks the absorption of sodium and chloride by the microvilli
and promotes the secretion of chloride and water by the crypt cells.
The result is watery diarrhea with electrolyte concentrations isotonic
to those of plasma
Vibrio sp.
• Vibrio parahemolyticus
• Endotoxin, hemolysin
• Seafood poisoning
• Vibrio vulnificus – ONPG + ; LF
• Endotoxin
• Seafood poisoning, wound infection, septicemia
• Vibrio alginolyticus
• Endotoxin
• Ear and wound infection
Plesiomonas shigelloides
• Endotoxin
• Ingestion, exposure to cold blooded animals
• Gastroenteritis, septicemia
Aeromonas hydrophilia
• Endotoxin
• Ingestion, skin penetration
• Gastroenteritis, wound infection, bacteremia, endocarditis,
conjunctivitis, pneumonia
Drugs of Choice:
• Vibrio – chloramphenicol, tetracycline, quinolones, erythromycin
• Plesiomonas and Aeromonas – beta lactam drugs; quinolones
Gram Negative Bacilli/Coccobacilli
(Aerobic)
• McConkey Agar Growth
• McConkey Agar No Growth
• Oxidase variable : Haemophilus
• Oxidase + : Sphingomonas paucimobilis, Moraxella, Neisseria elongata,
Eikenella corrodens, Weeksella virosa, Pasturella, Suttonella, Mannheimia
haemolytica, Actinobacillus, Kingella, Cardiobacterium, Capnocytophaga
• Growth Require Special Media
• Pseudomonas, Burkholderia, Achromobacter, Rhizobium, Ochrobactrum,
Chyseobacterium, Sphingobacterium, Alcaligenes, Bordetella (non pertussis),
Comamonas, Vibrio, Aeromonas, Plesiomonas, Chromobacterium
Non fermentative but oxidative; oxidase
positive, obligate aerobe, NLF on MAC
• Oppportunistic, nosocomial infectionis
• Achromobacter denitrificans – Nitrate reduction positive
• Alkaligenes faecalis – Nitrate reduction negative
Family Pasteurellaceae
• Genus Pasturella
• P. multocida
• Genus Haemophilus
• H. influenzae
• H. influenzae biogroup aegyptius
• H. ducreyi
• H. paraaprophilus
• H. parainfluenzae
• H. parahemolyticus
• Genus Actinobacillus
• A. actinomycetomcomitans
Gram (-) Rod, MAC (-), nonsaccharolytic
• Eikenella corrodens
• Normal flora of the mouth, upper respiratory tract, Git
• Corroding bacterium on BAP
• Bleach like odor
• Human bite infections, head and neck infections, SBE
Capnocytophaga
• C. gingivalis
• C. ochracea
• Fusiform bacilli with one rounded end and one tapered end ; occ.
Filamentous
• BAP: opaque, shiny nonhemolytic beige to yellow colonies and exhibit gliding
motility similar to swarming of Proteus
• Habitat: normal flora of human oral cavity
• Dx: periodontitis, bacteremia
Cardiobacterium
• C. hominis
• Pleomorphic, tear drop appearance
• BAP: small, slightly alpha hemolytic, smooth, round, glistening, opaque, +/-
pitting
• Habitat: normal flora of human upper respiratory tract
• Capnophilic
• Opportunistic endogenous infection
• Dx: endocarditis esp. in those with anatomic heart defects
• Rx: Penicillin