Escherichia coli

Some strains are overt pathogens

5 Diarrheogenic E. coli
Enteropathogenic E.coli (EPEC)
- infantile, mucoid, chronic diarrhea
- loss of microvilli
- bundle-forming pilus, intimin

Enterotoxigenic E. coli (ETEC)

- major cause of travelers and childhood diarrhea
- heat labile (LT) and heat stable (ST) enterotoxin that mediate
secretion of water & electrolytes into the bowel lumen
- high infective dose
- nonbloody, watery diarrhea

Escherichia coli

Enteroinvasive E. coli (EIEC)

- causes dysentery (necrosis, ulceration, inflammation of large
bowel; seen in young children
- non-motile, NLF, invades enterocytes (Shigella-like)
- Sereny (+), watery to bloody diarrhea

Enterohemorrhagic E. coli (EHEC)

- VTEC - verotoxin producer VTEC
- STEC serotoxigenic (toxin similar to Shiga toxin)
- ex. O157:H7 and 0157:NM; inflammation & bleeding of the
mucosa hemorrhagic colitis; HUS, hemorrhagic diarrhea, colitis
- sMac(-), MUG(-), no WBC unlike dysentery

EnteroAggregative/Enteroadherent E. coli (EAggEC)

Watery diarrhea; no blood nor WBCss

Inflammation with fever and abdominal pain

Escherichia coli
Extra intestinal infections:
- UPEC uropathogenic
- MNEC meningitis / sepsis associated
- 80% with K1 anitgen
Lab ID:
- Green metallic sheen on EMB
- LF (A/AG)
- IMVC + + - -

Klebsiella pneumoniae

Nosocomial pneumonia
Pneumonia in smokers and alcoholics
UTI, wound infections, bacteremia
Nursery outbreaks
Lab ID:
- moist, mucoid colonies
- LF (A/AG)
- IMVC - - + +
- motility(-)

Klebsiella granulomatis
Donovan bodies

Enterobacter spp.
Lab ID:
- colonies may resemble Klebsiella
- LF
- IMVC - - + +
- motility (+)

Citrobacter freundii
Colonies may resemble Salmonella
late LF (K/AG + H2S); ONPG(+)
IMVC - + - +
Deaminase (-)
Lysine decarb(-), urease(+)

Cronobacter sakazakii
Yellow pigment enhanced by incubation at 25 C
Formerly Enterobacter

Serratia marcescens
RT and UT infections
Outbreaks in burn units, nurseries

Lab ID:
- produces red pigment (prodigiosin)
- late LF (K/AG or K/A); ONPG (+)
- IMVC - - + +

Proteus spp.
P. mirabilis
P. vulgaris
Associated with UTI and kidney stones
Lab ID
- swarming colonies on nonselective media
- burned chocolate odor
- NLF (K/AG + H2S)
- deaminase(+)
- urease (+)

Proteus spp.
IMVC
P. mirabilis

-+vv

P. vulgaris

++--

Morganella morganii

NLF (K/AG)
IMVC + + - deaminase(+)
H2S (-)

Providencia spp.

NLF (K/A)
IMVC + + - +
deaminase(+)
H2S (-)

Edwardsiella tarda
NLF (K/AG + H2S)
IMVC + + - urease (-)

Review
LF
- E.coli
- Klebsiella
- Enterobacter
Late LF
- Citrobacter
- Serratia
NLF
- Proteus
- Morganella
- Providencia
- Edwardsiella

Non-motile
- Klebsiella
- Shigella
- Yersinia (temp dependent)
. H2S positive
- Citrobacter
- Proteus
- Edwardsiella

PADT positive
- Proteus
- Morganella
- Providencia

Salmonella
Salmonella enterica
- 7 subgroups with diff subspecies
Subgroup I
- S. typhi, S. paratyphi, S. choleraesuis
Subgroup II, III, IV
- zoonotic
- Arizona (subgroup III)

Salmonella
Infections:
Gastroenteritis
Typhoid fever
Bacteremia
Carrier state
Salmonella typhi
- NLF (K/A + H2S)
- IMVC - + - - Urease(-)
- Lysine decarb(+)

Special Media
- Sel-F
- XLD
- SSA
- Hektoen Enteric

3 general categories of infection:

Gastroenteritis and diarrhea (mucosa and
submucosa)
Bacteremia and extraintestinal infections
Enteric / typhoid fever

Shigella

Agents of dysentery
High infective dose (<200 bacilli)
Direct person-to-person transmission
NLF
Motility(-)
H2S (-)
Scientific names of Group A, B, C, D

Shigella
Shigella sonnei
- predominant isolate in the US
- late LF
Shigella flexneri
- gay bowel syndrome
Shigella dysenteriae type 1
- most virulent
- IMVC v + - -

Yersinia pestis
Agent of plague
2 forms:
a. bubonic / glandular
- vector borne: Xenopsylla cheopsis, Pulex
irritans
b. pneumonic
- usually following blood-borne dissemination
- aerosol transmission

Yersinia pestis
Lab ID
bipolar staining; safety pin
NLF
IMVC - + - motility (-)

Y. enterocolitica & Y.
pseudotuberculosis
Agent of pseudoappendicitis / mesenteric
adenitis
Enterocolitis
Zoonotic
Bipolar staining
Best recovered following cold enrichment
(4oC, 2-4 wks)
Y. enterocolitica motile at 25oC
Y. pseudotuberculosis motile at 18 - 22oC

Acinetobacter, Stenotrophomonas
Oxidase negative: Pseudomonas luteola,
Pseudomonas oryzihabitans &
Stenotrophomonas maltophila (formerly
Xanthomonas, Pseudomonas)
Either oxidize glucose (saccharolytic) or they
do not utilize glucose (asaccharolytic or non
oxidizers)
A. baumannii (hospital acquired,
asaccharolytic)

Other nonfermentative
gram negative bacilli - nosocomial
MAC: colorless, NLF
Motile with peritrichous flagella, obligate
aerobe, non fermentative but oxidative
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

MAC (+), oxidase (+),

glucose fermenters,
aerobic / facultative anaerobic
Family Vibrionaceae
Vibrio cholerae, parahemolyticus, vulnificus,
alginolyticus
Plesiomonas shigelloides

Family Aeromonadaceae
Aeromonas caviae
Aeromonas hydrophila

Vibrio

V. cholerae
V. parahemolyticus
V. vulnificus
V. alginolyticus
Limulus test detects endotoxins visible gel in
lysates of washed amebecytes of horse crab
Ileal loop test detects enterotoxins
Kamagawa phenomenon double zone hemolysis
Wagatsuma agar

Vibrio/Plesiomonas/Aeromonas
Colonial
morphology

TCBS sucrose
fermenter yellow
Non sucrose
fermen - blu/green

(-) TCBS
Non fermenter

Microscopic
morphology

G(-) curved, comma G(-) comma or

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

G(-) straight bacilli

motility

+ darting motility

(-) TCBS
Non fermenter

Vibrio/Plesiomonas/Aeromonas
habitat

Brackish or salt
water

Fresh water esp. in

warmer climate

Fresh, brackish
marine, polluted or
chlorinated water

Oxidase test

All +) except V.
metschnikovii

(+)

(+)

Halophilic (grows
on 6 % NaCl)

All (+) except V.

No
cholerae & mimicus

No

Growth at
pH 8.4 9.5

Will grow

Will not grow

Will not grow

Inositol
fermentation

Non fermenter

fermenter

Non fermenter

Vibrio/Plesiomonas/Aeromonas
Susceptibility to
vibriostatic agent
0/129

susceptible

susceptible

resistant

String test w/ 0.5 %

Na deoxycholate

positive

negative

negative

Vibrio spp.

Found in aquatic environments

halophilic
Small gram-negative rods
Curved in direct examination
Oxidase (+)
Susceptible to vibriostat disk (O/129)
Positive string test (0.5% Na desoxycholate)
Best cultivated in TCBS
Enrichment: APW w/ 1% NaCl

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
washerwomans hands, sunken eyes,
hypotension, death

Vibrio cholerae
Agent of cholera
(choleragenic strains: O1, O139)
- rice water stools
- clinical diagnosis is usually enough
Lab ID:
- yellow colonies in TCBS
- darting motility
- agglutinates in O1 anti-sera

El Tor

Classical

Hemagglutination
using chicken RBC

Hemolyis using
sheep RBC
Voges Proskauer

Phage IV suscep.

Polymxyin B
susceptibility

Vibrio sp.
Vibrio parahemolyticus
Endotoxin, hemolysin
Seafood poisoning

Vibrio vulnificus
Endotoxin
Seafood poisoning, wound infection, septicemia

Vibrio alginolyticus
Endotoxin
Ear and wound infection

Other Vibrio
V. cholerae non O1
- gastroenteritis
V. parahaemolyticus
- major cause of diarrhea in Japan
V. vulnificus
V. alginolyticus

Other Fermenting Gram-Negative

Bacilli
Aeromonas
- diarrhea, wound infections, bacteremia
- ice water, fish
- zoonotic
Plesiomonas shigelloides
- gastroenteritis, bacteremia, and meningitis
- cross agglutinates with Shigella sonnei

Aeromonas hydrophilia
Endotoxin
Ingestion, skin penetration
Gastroenteritis, wound infection, bacteremia,
endocarditis, conjunctivitis, pneumonia

Plesiomonas shigelloides
Endotoxin
Ingestion, exposure to cold blooded animals
Gastroenteritis, septicemia

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

Non-Fermenting Gram-Negative
Bacilli
TSI or KIA non-reactive (K/K, K/NC)

Pseudomonas / Burkholderia
Acinetobacter
Stenotrophomonas
Campylobacter
Helicobacter

Family Pseudomonaceae
Pseudomonas aeruginosa, fluorescens, putida,
stutzeri
Burkholderia cepacia, mallei, pseudomallei
Stenotrophomonas (Xanthomonas)
maltophilia
Comamonas testosteronii, Shewanella
putreficans, Brevundimonas diminuta,
Ralstonia pickettii, Ralstonia mannitolliytica

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
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
Oxidase (+), motile, grape/fruity odor, pigments
Slime layer, pili, O Ag, endotoxin, phosphorylase
(destroys pulmonary surfactant)
P. aeruginosa blue/green pus,swimmers rash
nosocomial infection, drug resistance, burns
P. mallei horses Glanders/Farcys disease
P. pseudomallei meliodosis (lung infection)
P. maltophila oxidase negative
Opportunistic P. stutzeri (brown pigment),
P.fluorescens, P. putida

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,
swimmers ear, Shanghai fever, jacuzzi or hot
tub syndrome, resp tract infection

Pseudomonas aeruginosa

Most common nonfermenter isolated

Nosocomial infections
Pneumonia in cystic fibrosis patients
Ear infections
Jacuzzi / hot tub syndrome
Burn infections
Lab ID:
-pyocyanin production
- fruity odor / mousy odor
- non-fermenter, oxidase(+), motility (+)

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

Stenotrophomonas maltophilia
Lavender green on BAP; yellow tan on TSA
Direct contact with hospital devices
Nosocomial infections

Opportunistic pathogens
Pigment

Oxidase

ONPG

Xanthomonas Yellow

(-)

(+)

Comamonas

None

(+)

(-)

Shewanella

Pink - brown

(+)

(+)

Other Pseudomonads
Burkholderia mallei
- Glanders

Burkholderia pseudomallei
- melioidosis / pseudoglanders
- Vietnamese time bomb