Escherichia coli

Submitted to
Dr.A.Roy
Professor& Head
Deptt.of microbiology
Anand veterinary college
Submitted by
Smita
M.V.Sc.scholar
Deptt.of vety.public health
Anand veterinary college
 Introduction
Escherischia coli is Gram negative
bacteria, usually motile with peritrichous
flagella and often fimbriate.
Lactose fermenter and produces pink
colonies on Macconkey agar and has
characteristic biochemical reactions in
IMViC tests.
Some strains produce colonies with a
metallic sheen when grown on eosin-
methylene blue agar.
Cont..
Organism persist as important members of
the normal flora of the intestine
throughout life.
Colonization of mammalian intestinal tract
by E.coli from environmental sources
occurs shortly after birth.
Somatic(O),flagellar(H)and capsular(K)
antigens are used for serotyping E.coli.
Cont...
Pathogenic strains of E.coli possess
virulence factors which allow them to
colonize mucosal surfaces and
subsequently produce disease.

In recent years, E.coli 0157:H7 has

emerged as a major food-borne, zoonotic
pathogen in humans.
 History
Escherichia coli was first isolated by
Escherich in 1885 from the faeces of infants
and he named the bacterium as “bacterium
colicommune”.
In 1920 Castellani and Chalmers gave the
genus name in honour of Escherich.
The organisms has been found in the
intestinal tract of practically all vertebrates.
The organisms under favourable
conditions ,undergo rapid multiplication and
cause disease in animals and humans.
 Habitat
Widely distributed in nature

Commonly present in large intestine

of man and animals.
 Morphology
E.coli is a short rod measuring 2-3 µm in
length and 0.6µm in width.
Does not form spore, motile due to
peritrichous flagella and often fimbriate.
Some strains produces a polysaccharide
capsule and produces a mucoid growth.
The organism is readily stained and is
Gram negative.
Unstained cells of E. coli viewed by phase microscopy. about 1000X magnification.
Gram stain of E. coli viewed by conventional light microscopy. about 1000X magnification. .
E.Coli with fimbriae
 Growth requirement
 The organism is aerobic and
facultatively anaerobic.
 optimum temperature of growth is

37°C but growth occurs over a range

of 14-45°C.
 The optimum pH of growth is 7.0 but

organism grows within a wide pH

range.
 Cultural characteristics
The organism grows well on ordinary media
and form large colonies of 2-3 mm in
diameter.
The colonies are circular, low, convex,
smooth and colourless in 18 hours of
incubation on nutrient agar.
On MacConkey’s lactose agar, the growth
appears as large pink coloured colonies due
to fermentation of lactose and production
of acid shown by neutral red indicator.
Cont..
Haemolytic activitiy on blood agar is
characteristic of certain strains of E.coli
In broth the smooth strains produce
diffuse cloudiness and a heavy sediment
but no pellicle.
When grown on eosin-methylene blue
agar(EMB agar), some strains produce
colonies with a metallic sheen.
Growth of E.coli on MacConkey agar
Growth of E.coli on EMB agar
Biochemical characteristics
E.coli produces acid and gas from glucose,
lactose, fructose ,galactose, maltose and
mannitol, may or may not ferment sucrose,
salicin and dulcitol.
Methyl red positive
Voges-Proskauer negative,
Forms indole,
reduces nitrates to nitrites,
does not produce H2S in TSI(triple sodium
ion)
Cont..
 Does not grow on citrate medium.
 Do not show urease activity.

 Shows lysine decarboxylase activity.

 Clinical relevance, growth characteristics,& biochemical reactions of members of enterobacteriaceae

Escherichia coli Salmonella Yersinia sps. Proteus sps. Enterobacter Klebsiella

aerogenes pneumoniae

Clinical Major pathogen Major Major Opportunistic Opportunistic Opportunistic

importance pathogen pathogen pathogens pathogens pathogens

Cultural Some strains _ _ Swarming growth mucoid mucoid

characteristics haemolytic

Motility at 30◦C motile Motile Motile Motile Motile Non-motile

Lactose + _ _ _ + +
fermentation

IMVIC tests

Indole + _ v ± _ _
production

Methyl red test + + + + _ _

Voges-proskauer _ _ _ v + +
test

Citrate utilization _ + _ v + +
test

H2S production in _ + _ + _ _
TSI agar

Lysine + + _ _ + +
decarboxylase
 Antigenic character
Somatic(O),flagellar(H),and capsular(K)
antigen are used for serotyping E.coli.
Somatic antigen are lipopolysaccharide in
nature and located at surface of cell wall
Flagellar antigen are protein in nature.
Capsular antigen are composed of
polysaccharides.
Proteinaceous fimbrial(F) antigens act as
adhesins facilitating attachment to
mucosal surfaces.
Pathogenesis and pathogenicity
Virulence factors of pathogenic strains of E.coli include
capsules, endotoxins, structures responsible for colonization,
enterotoxins & other secreted substances.

Capsular polysaccharides, interfere with phagocytic

uptake of E.coli. Capsular material which is weakly antigenic,
also interferes with antibacterial effectiveness of complement
system

Endotoxin, a lipopolysaccharide(LPS) component of cell

wall of Gram-negative organism,is released on death of
bacteria.It is composed of lipid A moiety, core polysaccaride
and specific side chains.

Role of LPS in disease production includes pyrogenic

activity, endothelial damage, endotoxic shock.these effects
are of great significance in septicaemic disease.
Cont..
Fimbrial adhesins(present on many
enterotoxigenic strains of E.coli )allow attachment
to mucosal surfaces in small intestine and in lower
urinary tract.

Most significant adhesins in strains of E.coli

producing disease in domestic animals are
K88(F4),K99(F5),987P(F6)and F41.

Most common adhesin present in strains of E.coli

infecting pigs is K88.

K99 &F41 adhesins occur in calves & K99 in lambs.

Cont..
Both K88 &K99 adhesins are encoded by
plasmids.

An adhesin, termed intimin,appears to be

necessary for binding of enteropathogenic
E.coli(EPEC) to enterocytes
E.coli on surface of small intestine
Bladder epithelial cells with attached E.coli cells to it
Cont..
Pathological effects of infection with
pathogenic E.coli derive mainly from:
• Endotoxin
• Enterotoxin
• Verotoxin
• Cytotoxic necrotizing factors
Enterotoxin affect only functional activity of
enterocytes while verotoxins and cytotoxic
necrotizing factors can produce
demonstrable cell damage at their sites of
action.
 pathogenic
strains of E.coli

non-enteric
enteric disease septicaemia
localized disease

invasion by
enterotoxigenic enteropathogeni septicaemic uropathogenic
oppurtunistic
E.coli(ETEC) c E.coli(EPEC) strains of E.coli strains of E.coli
E.coli

endotoxin
enterotoxins induced tissue
changes

heat-labile(LT)

heat- stable(ST)

verotoxigenic necrotoxigenic
E.coli(VTEC) E.coli

verotoxins:VT1,V cytotoxic necrotizing

T2,VT2e factors:CNF1,CNF2
Cont..
 Enterotoxigenic E.coli

Adhesins,K88 in pigs,K99 in calves and in

lambs,required for colonization
LT1 induces hypersecretion of fluid into intestine through
stimulation of adenylate cyclase activity.
LT1 has been demonstrated in many strains of ETEC.
LT2 in some strains of ETEC isolated from cattle
STa induces increased guanylate cyclase activity in
enterocytes and this inhibit fluid absorption from intestine
Precise cytotoxic effect of STb is not known.
Cont..
 Enteropathogenic E.coli(EPEC):
An adhesin intimin, required for colonization
Cause destruction of microvilli,atrophy and
shedding of enterocytes
Maldigestion, malabsorption
Cause diarrhoea in piglets, lambs, pups

 Verotoxigenic E.coli(VTEC):
Bind to enterocytes
Damage to vasculature in intestine and in other
locations
Oedema disease in pigs
Post-weaning diarrhoea in pigs
Cont..
 Necrotoxigenic strains of E.coli:
Bind to enterocytes
Damage to enterocytes and to blood
vessels
Cause haemorrhagic colitis in cattle
 Term Enterohaemorrrhagic E.coli is
applied to strains such as O157:H7
which cause dysentry in humans.
Cont..
Alpha-haemolysin,are useful marker for
virulence in certain strains of E.coli,is closely
related to expression of other virulence
factors.
Siderophores, iron-binding molecules such as
aerobactin and enterobactin,are synthesized
by certain pathogenic strains of E.coli.
When available iron levels in tissues are low,
these iron binding molecules may contribute
to bacterial survival
 Clinical infections
In young animals
Enteric colibacillosis(calves,lambs,piglets)
Neonatal diarrhoea
Colisepticaemia(calves ,lamb and poultry)
Systemic colibacillosis
Colibacillary toxaemia
In older animals
Post-weaning enteritis(pigs)
Oedema disease(pigs)
Non-enteric localized infections which involve
urinary tract, mammary glands and uterus
 Diagnosis
Suitable specimens include
Faecal samples from enteric cases
Tissue from septicaemic cases
Mastitic milk
Samples of mid-stream urine and
Cervical swabs from suspected cases of
pyometra or metritis
Specimens cultured on blood and
MacConkey agar are incubated aerobically
at 37оC for 24 to 48hrs.
Cont..
On blood agar the colonies are greyish, round
and shiny with a characteristic smell.colonies
may be haemolytic or non-haemolytic.
On MacConkey agar colonies are bright pink.
IMViC tests can be used for confirmation
Colonies of some E.coli strains have a
metallic sheen on EMB agar.
Slide agglutination tests for O and H antigens
are employed for serotype identification
Cont..
When enterotoxigenic strains of E.coli are
suspected, presence of either enterotoxins or
fimbrial antigens can be confirmed by
immunological methods or molecular
techniques such as PCR.

Enterotoxin in small intestine can be detected

using methods employing monoclonal
antibodies.

For expression of fimbrial antigens, isolates

should be subcultured on Minca
medium.Fimbrial antigens can be identified
using ELISA or Latex agglutination
Cont..
DNA probes specific for genes encoding
heat-labile and heat-stable enterotoxins
may be used to identify enterotoxigenic
strains of E.coli.
Toxins produced by verotoxigenic and
necrotoxigenic strains can be detected by
vero cell assay.
Molecular methods based on the detection
of genes encoding toxins are also used.
 Treatment
In calves with neonatal diarrhoea,milk
should be withdrawn and replaced by fluids
containing electrolytes.
Milk feeding can be resumed gradually
when clinical improvement is evident.
Enteric diseases may be treated by oral
administration of antimicrobial compounds
which are active in GI tract
Systemic and localized infections require
parenteral administration of therapeutic
agents.
 Prevention&Control
Newborn animals should receive ample
amounts of colostrum shortly after birth.
Colostrum antibodies can prevent
colonization of intestine by pathogenic
E.coli.
Clean, warm environment should be
provided for newborn animals
Dietary regimes may contribute to the
development of oedema disease and other
post-weaning conditions.
New feed should be introduced gradually.
Cont..
Vaccination methods used for prevention of
enteric disease in piglets and calves include:
Commercially available killed vaccines
containing prevalent pathogenic E.coli
serotypes can be given to pregnant sows.
Vaccination of pregnant cows with purified
E.coli K99 fimbrial or whole-cell
preparations,often combined with rotavirus
antigen,can be used to enhance colostral
protection