E.
COLI
Morphology: Gram -ve, rod shaped. It is 1-3 x 0.4-0.7 µm in size and
0.6 to 0.7 µm in volume. It is arranged singly or in pairs. It is motile due
to peritrichous flagella, though some strains are non-motile. Spores are
not formed. Capsules (polysaccharide) and fimbriae are found in some
strains.
Two types –
S (smooth) forms
R (Rough) forms
S – forms are seen in fresh isolates, easily emulsifiable in
saline.
R – forms are seen in older cultures, with irregular dull
surface, often auto-agglutinable in saline.
S-R variation occurs as a result of repeated subcultures and is
associated with the loss of surface antigens and usually of
virulence
Growth & Cultural characteristics:
It is an aerobe/ facultative anaerobe. The optimum growth temperature is
37°C.
On Nutrient agar, colonies are large, thick, greyish white, moist, smooth,
opaque or translucent discs. Some strains may form “mucoid ” colonies.
On MacConkey agar medium, colonies are bright pink due to lactose
fermentation.
On selective media, their growth is inhibited, however their colonies are
pink on DCA (Desoxycholate citrate agar) as it contains lactose and
neutral red. In broth, there is generalized turbidity and deposit which
disperses on shaking.
Biochemical Reactions:
Glucose, lactose, mannitol, maltose are fermented with acid and gas
production, but sucrose is not fermented by typical strain of E. coli. In
Triple sugar iron (TSI), acid and gas are produced.
Indole = +
MR (methyl red) = +
Nitrate reductase = +
VP (Voges Proskauer) = -ve
oxidase = -ve
citrate negative = -ve
H2 S is not formed and urea is not hydrolysed.
Epidemiology patterns and Transmission:
The most serious illness in humans is caused by verocytotoxic
E. coli (VTEC) (also known as enterohaemorrhagic E. coli), The most
common VTEC strain in Europe and North America is E. coli O157:H7.
However, other serotypes have frequently been involved in sporadic cases
and outbreaks.
Important cause of bloody diarrhoea in Europe.
In 2005 there were 946 laboratory confirmed cases of VTEC 0157 in
England and Wales (HPA).
The highest incidence rates in the UK are seen in children < 5
years.
The highest rates have been observed in rural areas particularly in
Scotland
An estimated 73,480 illnesses due to E. coli O157 infection occur
each year in the United States, leading to an estimated 2,168
hospitalizations and 61 deaths annually, and it is an important
cause of acute renal failure in children.
Mode of Transmission:
Primarily through the consumption of contaminated, undercooked or
raw foods, (particularly ground beef) and unpasteurised milk.
Other foods implicated in outbreaks of [Link] O157:H7 include
undercooked hamburgers, dried cured salami, lettuce, yogurt,
cheese and milk and radish sprouts.
Cross contamination during food preparation.
Person to person via the faeco-oral route is common.
Direct contact with animals, e.g. school visits to farms.
Waterborne transmission occurs through swimming in or consuming
contaminated water.
The infectious dose of VTEC O157 appears to be very low, probably
less than 100 organisms.
Pathogenicity:
The most widely distributed clone is E coli O25b/ ST131
Antigens:
Somatic O Ag - 164 – Endotoxic activity
Capsular K Ag - 100
Flagellar H Ag - 50
Antigenic determinants:
• Fimbriae (2 types)
Mannose sensitive (common pili)
Mannose resistant
• K1 - Capsular Antigen – protects against phagocytosis
Enterotoxin:
LT- Heat Labile Toxin
o It is due to increase in In CAMP causes to increase secretion of NA+
& Cl- and water from cells.
ST- Heat Stable Toxin
o Activation of Cyclic guanosine monophosphate and
Increased cGMP.
o Causes fluid accumulation in intestine
Some strains produce Verotoxin: Vt1, Vt2- Also known as
shiga toxin-producing acts like shigella dysentery toxin and
causes cytotoxicity.
Hemolysin- cytotoxic and facilitate invasion.
Virulence Factors:
Virulence factors of E. coli can affect a wide range of eukaryotic
cellular processes, including cell signalling, ion secretion, protein
synthesis, mitosis, cytoskeletal function and mitochondrial function.
Virulence factors of pathogenic E. coli are frequently encoded on
genetic elements such as plasmids, bacteriophage, transposons and
pathogenicity islands that can be mobilized into different strains to
create novel combinations of virulence factors.
Virulence Factors:
1) LT,ST, Colonizatrion factors – present in Enterotoxigenic
(ETEC) [Link] and causes Watery to cholera-like diarrhea.
2) Ipas, VirG/IcsA factors – Present in Enteroinvasive (EIEC)
[Link] and causes Watery diarrhea to dysentery
3) Intimin, Tir (Bundle forming Pilus), BFP factors – present in
Enteropathogenic (EPEC) [Link] and causes Watery diarrhea
(infantile diarrhea)
4) EPEC factors + shigatoxins, EspP and hemolysin - present in
Enterohemorrhagic (EHEC) (O157:H7) [Link] ( shiga toxin-
producing) and causes Hemorrhagic cystitis, Hemolyic urimic
syndrome (HUS)
5) AAF adhesins, EAST-1, Pet, Pic, Hemolysin - present in
Enteroaggregative (EAEC) [Link] and causes watery to mucoid
diarrhea
6) Adhesins and Esp-like proteins - present in Diffusely
adhering (DAEC) [Link] and causes watery diarrhea
7) Type-1 pili, P -pili, Afa, hemolysin, CNF-1 - Present in
Uropathogenic (UPEC) [Link] and causes Urinary tract
infections (UTI’s)
8) Capsule, Type-1 pili, S-fimbrial adhesin, IbeA and IbeB
invasins - Present in Septic (SEC) [Link] and causes Neonatal
sepsis and meningitis.
PATHOGENEIS: ENTEROHEMORRHAGIC (ECOLI ) it occurs in large intestine
which is due to presence of shiga like toxins similar to shiga like toxins and
cholera . O157:H7 is the most [Link] has two types of toxins shiga like
toxin 1 Shiga like toxins 2 urinarination a severe form of diarrhea, and with
hemolytic uremic syndrome, a disease resulting in acute renal failure,
microangiopathic hemolytic anemia, and thrombocytopenia. It also has the
heat labile and heat stable toxins. HLT increases the CAMP which is secondary
messenger of cytoplasm which activates the ion channels and this causes to
activate the intracellular electrolyte like chloride and water channels to go out
of the cells that is to the external environment. In this way they promote the
diarrhea and inhibition of the reabsorption of sodium... The gut lumen is
distended with fluid, and hypermotility and diarrhea ensue, lasting for several
days. LT is antigenic and cross-reacts with the enterotoxin of Vibrio cholerae,
since it also has the similar mechanism. LT stimulates the production of
neutralizing antibodies in the serum (and perhaps on the gut surface) of persons
previously infected with enterotoxigenic E coli. For the HEAT STABLE
TOXIN there will be an increase of CGMP which leads to the activation of
protein kinase which controls the ion channels and when there is an opening of
ion channels this leads to the efflux of water into the lumen which promotes
secretory diarrhea. Ecoli has the ability to produce the verotoxin and it causes
the B- HEMOLYSIS because of the hemolysin and it causes to kill the normal
cell(cytotoic) and it also facilitates [Link] also because of the
colonization factors such as fimbrae which is helpful for attachment and for this
we have mannose resistant (pilli) and the mannose sensitive which atteches to
the intestine to resist peristalsis. And also adherenece to vaginal epithelial cells
and may contribute to vaginal colonization and cystitis.
FOR ONLY THE REFERENCE PURPOSE WHY IT IS ONLY ECOLI
HAEMORRAGIC:
We totally have 6 forms of strains which cause diarrhea but among that for only
4 strains it causes the bloody diarrhea ETEC, EPEC, EHEC, EIEC. Among all
this there will be a enterotoxins such as heat labile and heat stable. But only for
the haemorrogic colitis we have the shiga like toxin, verotoxin and the
hemolysis [Link] is responsible for the acute kidney injury and bloody
urination and also symptoms mentioned to the patient( PLEASE REFER THE
VIRULENCE FACTORS MENTIOED IN PATHOGENICITY)
CLINICAL FEATURES OF THE DISEASE CAUSED BY ORGANISM:
[Link] will be a cystitis/ pyelonephritis which is due to uropathogenic ecoli.
[Link]
3. Accumulation of pus in pleural fluid.
4. peritonitis because of the rupture of appendix.
5. sepsis
6. neonatal meningitis.
Clinical presentation of the patient that is correlated with the organism’s characteristic: for the patient it was
mentioned there will be a blood in urine, bloody diarrhea, acute kidney injury, abdominal pain, vomiting and
dehydration.
DIAGNOSIS: ; O157:H7 strains also are negative on MUG tests . Specific anti sera are used to identify
the O157:H7 strains. Tests for the detection of both Shiga toxins using commercially available enzyme
immunoassays (EIAs) are done in many laboratories. Other sensitive test methods include cell culture cytotoxin
testing using Vero cells and polymerase chain reaction for the direct detection of toxin genes directly from stool
samples. Specimens: include urine, blood, pus, spinal fluid, sputum, or other material, as
indicated by the localization of the disease process.
Smears: The Enterobacteriaceae resemble each other morphologically. The presence of
large capsules is suggestive of Klebsiella species.
Culture: Specimens are plated on both blood agar and differential media. With differential
media, rapid preliminary identification of gram-negative enteric bacteria is often possible.
Nucleic Acid Amplification Tests (NAATs)
A variety of multiplex NAATs designed to detect the most common pathogens
responsible for particular syndromes, are currently available and many more
are entering clinical trials.
TREATMENT:
1. Trimethoprim- Sulfamethoxazole
2. Ciprofloxacin
3. sulfonamides, Ampicillin, cephalosporins, fluoroquinolones,
and aminoglycosides which
have marked antibacterial effects against the Enterics.
Multiple drug resistance is common and is under the control of
transmissible
Plasmids.
Fluid and electrolyte replacment.
PREVENTION: Cooking ground beef and by avoiding unpasteurized
products such as apple cider.
1. Avoid raw milk
2. washing hands.
3. Maintain sanitation
Developing farm and slaughterhouse-based methods to decrease
contamination of meat; encouraging use of irradiation to increase
the safety of ground beef; identifying ways to prevent
contamination of foods eaten raw (e.g., produce).
Follow correct food hygiene practices for food preparation and
cooking in domestic and commercial kitchens as described by the
WHO Five keys to safer food3.
These include;
Prevent cross contamination of raw and cooked food by washing
hands before, during and after food preparation. Wash and
sanitize all equipment, surfaces and utensils used for food
preparation.
Separate raw and cooked food, and use separate equipment and
utensils for handling raw food.
Cook food thoroughly (until centre of food reaches at least 70 oC),
especially poultry, meat, eggs and seafood.
Reheat cooked food thoroughly, and store cooked and raw food at a
safe temperature.
Use safe water and raw materials, e.g. pasteurized milk and water.
Wash fruit and vegetables.