ENTEROBACTERIACEAE

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Characteristics of the Enterobacteriaceae
 Gram-negative bacilli

Gram-negative bacilli
Ferment glucose
Reduce nitrates to nitrites
Oxidase negative
Catalase positive (except Sh.disentry type 1)
All except Klebsiella, Shigella and Yersinia are
motile by peritrichous flagella
 Grow readily on
 MacConkey (MAC) agar
 Eosin methylene blue (EMB) agar

 Grow readily at 35oC except Yersinia (25o-30oC)

 Do not form spores

 Natural Habitat:
 Environment (soil, water, and plants)
 Intestines of humans and animals
Enterobacteriaceae Meningitis

Opportunistic pathogens Pneumonia

Escherichia coli
Klebsiella pneumoniae Sepsis
Enterobacter aerogenes
Serratia marcescens
Diarrhea
Proteus spp.
Providencia spp.
Citrobacter spp. UTI

Obligate pathogens
Salmonella spp.
Shigella spp.
Yersinia spp.
Some E. coli strains
Classification of Enterobacteriaceae:
- Based on lactose fermentation

 LF
 LLF
 NLF
Classification of Enterobacteriaceae:
 Very large number of organisms in Family
Enterobacteriaceae, species are grouped into - TRIBES

 Tribe 1 – Escherichieae
 Tribe 2 – Klebsiellae
 Tribe 3: Proteae
 Tribe 4 : Yersiieae
 Tribe 5 : Erwinieae

 Within each Tribe, species are further subgrouped under

genera
Major Genera of family Enterobacteriaceae:

1. Escherichia

2. Shigella

3. Salmonella

4. Yersinia

5. Klebsiella

6. Proteus
Antigenic Factors of Enterobactericeae:

 Ability to colonize, adhere, produce various toxins and

invade tissues

 Some possess plasmids that may mediate resistance to

antibiotics

 Imp antigens used to identify the organisms:

 O antigen – somatic, cell wall, heat-stable Ag
 H antigen – flagellar, heat labile Ag

 K antigen – capsular, heat-labile Ag

Clinical Significance of Enterics:

 Ubiquitous in nature

 Except for Salmonella, Shigella and Yersinia, most are

present in the intestinal tract of animals and humans as
commensal flora; “fecal coliforms”

 Some live in water, soil and sewage

Clinical Significance of Enterics:

Based on clinical infections produced, enterics are

divided into two categories:
 Opportunistic pathogens – normally part of the usual
intestinal flora, that may produce infection outside the
intestine
 Primary intestinal pathogens – Salmonella, Shigella, and
Yersinia sp.
Enterobacteriaceae: Modes of Infection

1. Contaminated food and water (Salmonella spp.,

Shigella spp., Yersinia enterocolitica, Escherichia coli
O157:H7)

2. Endogenous infection (urinary tract infection, primary

bacterial peritonitis, abdominal abscess)

3. Abnormal host colonization (nosocomial pneumonia)

Transfer between debilitated patients

4. Insect (flea) vector: unique for Yersinia pestis

1. Urinary tract infection: Escherichia coli, Klebsiella
pneumoniae, Enterobacter spp., and Proteus mirabilis

2. Pneumonia: Klebsiella pneumoniae; Enterobacter spp.,

3. Wound Infection: Escherichia coli, Enterobacter spp.,

Klebsiella pneumoniae, and Proteus mirabilis

4. Bacteremia: Escherichia coli, Enterobacter spp.,

Klebsiella pneumoniae, and Proteus mirabilis
Enterobacteriaceae: Intestinal Infection

 Escherichia coli

 Shigella

 Salmonella
 Yersinia enterocolitica
Identification of Enterobacteriaceae:
 IMViC Test

 Indole, Methyl Red, Voges-Prosakaur, Citrate

(IMViC) Tests:

 The IMViC series of reactions allows for the

differentiation of the various members of family
Enterobacteriaceae.
A) Indole –Positive, B) MR –Positive
C) VP- Positive, D) Citrate- Negative
Methyl Red-Voges Proskauer (MR-VP) Tests
Principle
Glucose

Acidic pathway Or Neutral pathway

Acety methyl carbinol

(ACETOIN)
Mixed acids
 pH less than 4.4
Barrit’s A
Methyl Red Barrit;s B
indicator
MR positive VP positive
Pink color
Red color E. coli
Klebsiella
 MR/VP test

Methyl Red test Voges-Proskauer test

 Red: Positive MR (E. coli)  Pink: Positive VP (Klebsiella)

 Yellow or orange: Negative MR (Klebsiella)  No pink: Negative VP (E. coli)

 Result
Example Reaction on TSI
Result Slant Butt
H2 S
color color
Non fermenter
Alk/Alk/-
e.g. Negative Red Red
(No action on sugars)
Pseudomonas
LNF A/Alk/- Negative
e.g. Shigella (Glucose fermented Red Yellow
without H2S)
LNF Positive
e.g. Salmonella & A/Alk/+ black in
Proteus (Glucose fermented butt Red Yellow
with H2S)

LF
A/A/-
e.g. E. coli, Negative
(three sugars are Yellow Yellow
Klebsiella,
fermented)
Enterobacter
TSI Reactions of the Enterobacteriaceae
 A/A + g = acid/acid plus gas (CO2)
 A/A = acid/acid
 A/A + g, H2S = acid/acid plus gas, H2S
 Alk/A = alkaline/acid
 Alk/A + g = alkaline/acid plus gas
 Alk/A + g, H2S = alkaline/acid plus gas, H2S
 Alk/A + g, H2S (w) = alkaline/acid plus gas, H2S (weak)
A/A + g
 Escherichia coli
 Klebsiella

A/A + gas, H2S

 Citrobacter freundii
 Proteus vulgaris
 Non-lactose, sucrose fermenter
Alk/A
 Shigella
 Providencia
Alk/A + g
 Salmonella serotype Paratyphi A
Alk/A + g, H2S
 Salmonella
 Proteus mirabilis
 Edwardsiella tarda
 Lactose fermenting
colonies

Non lactose fermenting

colonies
Escherichia coli
 Most significant species in the genus
 Important potential pathogen in humans
 Common commensal in the intestine
 Pink (lactose positive) colony with surrounding pink area
on MacConkey
 Ferments glucose, lactose, trehalose, & xylose

 Usually motile

 Positive indole and methyl red tests

 Does NOT produce H2S

 Simmons citrate negative

 Voges-Proskauer test negative

Pathogenesis and clinical diseases
Sepsis
In people with inadequate host defenses, e.g. the newborns.
Usually originates from UT or GI infections. Some infections
may be endogenous.
Meningitis
E. coli
(particularly K1
strains) and
S. agalactiae are
the leading causes Bactere
mia
of meningitis in
infants.
Urinary tract infection

E. coli is the most common cause of urinary tract infection.

Community- vs. hospital-acquired UT infection

Most infections originate from colon; the bacteria

contaminate the urethra, ascend into the bladder, and may
migrate into the kidney or prostate.

Symptoms: urinary frequency, dysuria, hematuria, and

pyuria. Can result in bacteremia and sepsis.

Uropathogenic E. coli strains produce P (Pyelonephritis-

associated) pili, which is associated with renal colonization
and may induce protective immunity, and hemolysin HlyA.
Infections caused by E.coli:

 Meningitis, gastrointestinal, urinary tract, wound, and

bacteremia
Gastrointestinal Infections:
- diarrhoegenic E. coli
1. Enteropathogenic (EPEC)
2. Enterotoxigenic E. coli (ETEC)
3. Enteroinvasive E. coli (EIEC )
4. Enterohemorrhagic E. coli (EHEC)
5. Enteroaggregative E. coli (EAEC)
1. Enteropathogenic (EPEC)
 Primarily in infants and children;
 Outbreaks in hospital nurseries and day
care centers
 Stool has mucous but not blood
 Identified by serotyping
Enteropathogenic E.coli

Destruction of surface microvilli

 Fever Gut lumen

 Diarrhea
 Vomiting
 Nausea
 Non-bloody stools
Enterotoxigenic E. coli

• Diarrhea resembling cholera

• Travelers diarrhea
 Enterotoxigenic E. coli
 Heat labile toxin
 Like cholera toxin
 Adenyl cyclase activated
 Cyclic AMP
 Secretion water/ions

 Heat stable toxin

 Guanylate cyclase activated
 cyclic GMP
 uptake water/ions
Enteroinvasive E. coli (EIEC )

Dysentery
- Resembles shigellosis
Enterohemorrhagic E. coli
 Usually O157:H7

Flagella Transmission electron

micrograph
 O157:H7 transmitted- through meat products or sewage-
contaminated vegetables
 Hemorrhagic diarrhea
 Bloody, copious diarrhea
 Few leukocytes
 Afebrile
 Hemolytic-uremic syndrome (HUS)
 Hemolytic anemia
 Thrombocytopenia (low platelets)
 Kidney failure
 Enterohemorrhagic E. coli

 Vero toxin- “shiga-like”

 Hemolysins

Enteroaggregative (EaggEC)
Cause diarrhea by adhering to the mucosal surface of the
intestine; watery diarrhea; symptoms may persist for over
two weeks
Urinary Tract Infections
 E. coli is most common cause of UTI and pyelo-
nephritis in humans
 Usually originate in the large intestine
 Able to adhere to epithelial cells in the urinary tract
Septicemia & Meningitis

 Most common causes of septicemia and meningitis

among neonates

 Acquired in the birth canal before or during

delivery

 E. coli also causes bacteremia in adults, secondary

to genitourinary tract infection or a gastrointestinal
source
 Klebsiella, Enterobacter
 Usually found in intestinal tract

 Wide variety of infections, primarily pneumonia, wound,

and UTI
 General characteristics:
 Non-motile

 Simmons citrate positive

 H2S negative, Weakly urease positive

 MR negative; VP positive
 Usually found in Gastro-intestinal tract

 K. pneumoniae is most commonly isolated species

 Possesses a polysaccharide capsule - which protects

against phagocytosis and antibiotics

 Makes the colonies moist and mucoid

 Frequent cause of nosocomial pneumonia
 Significant biochemical reactions

 Lactose positive

 Most are urease positive

 Non-motile
 Nosocomial pneumonia: Spread by health care
personnel and equipment

 Frequently caused by K. pneumoniae

 Often seen in middle-aged males who abuse alcohol

 Difficult to diagnose due to commensals in sputum
Proteus, Morganella & Providencia species
 All are normal intestinal flora

 Opportunistic pathogens
 Deaminate phenylalanine

 Non lactose fermenters

Proteus species
 P. mirabilis and P. vulgaris are widely recognized
human pathogens

 Isolated from urine, wounds, and ear and bacteremic

infections

 Both produce swarming colonies and have a distinctive

“burned chocolate” odor

 Both are strongly urease positive

 Both are phenylalanine deaminase positive

 Exhibits characteristic “swarming”
Laboratory Diagnosis of Enterics
Specimen collection:
 Specimens collected and transported in Cary-Blair,
Amies, or Stuart media

Isolation and Identification

 Site of origin must be considered
 Enterics from sterile body sites are highly significant

 Routinely cultured from stool

Media for Isolation and Identification of
Enterics:

 Blood agar and a selective/differential medium such

as MacConkey

 On MacConkey, lactose positive are pink; lactose

negative are clear and colorless
  For stool, highly selective media, such as Hektoen
Enteric (HE), XLD, or SS is used along with
MacConkey agar

 Identification: All enterics are

 Oxidase negative

 Ferment glucose
 Reduce nitrates to nitrites