Escherichia 1. E.

Coli
2. 'Alkalescens-Dispar' group (atypical forms

ofEsch. coli.) Morphology Strains ofEsch. coliare Gram Negative Non Spore forming Fimbriated 80% May form loose slime May produce a polysaccharide capsule Motility by peritrichous flagella

Culture Characteristics
They grow well on non-selective media Usually (with the exception of certain

Verocytotoxin-producing strains) ferment lactose Producing large red colonies on MacConkey agar. They grow over a wide range of temperature (15-45C) Some strains are more heat resistant than other members of the Enterobacteriaceae and may survive 60C for 15 min or 55C for 60 min.

MacConkey agar: Ferment lactose rapidly on MacConkey agar Colonies are rose pink in colour 1-1 Jam colony DCA growth inhibited Blood Agar: 1-3mm colony Some loose zone of hemolysis Entire edge low convex smooth and shiny

Esch. colican be differentiated from other

enteric Gram-negative bacteria by

Ability to utilize certain sugars Indole production Formation of acid and gas from lactose and

other carbohydrates

The termcoliformis commonly used to refer

to any member of the Enterobacteriaceae, but prefer to reserve it for enterobacteria that normally ferment lactose

Identification Tests
Sugar Fermentation
Glucose Lactose Sucrose Mannitol

+ + +

Bio Chemical Tests Indole Methyl Red Voges Proskauer Citrate Urease H2S production
IMVIC

Positive Positive Negative Negative Negative Negative + + - -

Antigenic Structure
Serotyping is based on the distribution of Lipopolysaccharide (LPS) Somatic (O) antigens Flagellar (H) Capsular (K) antigens

O antigen More than 180 different O antigens have been described and new 'O' groups continue to emerge. Serotyping detects cross-reactions as a result of shared epitopes on the LPS expressed by strains ofEsch. Coli Sometimes it gives false positive Widal test because of its shared epitopes

H antigen More than 50 H antigens have been identified. Most are monophasic, but rare diphasic strains have been reported. K antigen (100 K antigen Types) The term 'K antigen' was first used collectively for surface or capsular antigens that prevent flagellarspecific antibodies from binding to the somatic antigens. In the past these antigens were divided into three classes (L, A and B) according to the effect of heat on the agglutinability, antigenicity and antibody-binding power of bacterial strains that express them. In modern usage, 'K antigen' refers to the acidic polysaccharide capsular antigens, and those ofEsch. colimay be divided into two groups groups I and II

Fimbrial antigens Like many other members of the Enterobacteriaceae,Esch. colimay produce fimbriae, and strains may express both sex pili and more than one type of fimbrial structure. Type 1 fimbriae can mediate adhesion to a wide range of human and animal cells that contain the sugar mannose. Such adhesion might be involved in pathogenicity of GI and Urinary tract infections Fimbriae that are of importance in urinary tract infection cause mannose-resistant haemagglutination.. These include the P fimbriae that bind specifically to receptors present on the P blood group antigens of human erythrocytes and

Pathogenesis
E. coli is most abundant aerobic and

facultative anaerobe bacteria present in the gut It may cause infections from
self gut flora Contaminated sewage Nosocomial infections

Strains ofEsch. colipossess a range of

different pathogenic mechanisms. The polysaccharides of the O and K antigens protect the organism from the bactericidal effect of complement and phagocytes in the absence of specific antibodies.

Many strains express haemolysin(s),

Strains ofEsch. coliisolated from human extra-

intestinal infections are more likely to be haemolytic than strains isolated from the faeces of healthy human beings.

Haemolysin production is an important

pathogenic mechanism for releasing essential ferric ions bound to haemoglobin, and the expression of certain haemolysins has been shown to be regulated by iron.

The ability of strains ofEsch. colito acquire

ferric ions is a recognized pathogenic mechanism.

Expression of the aerobactin-mediated iron-

uptake system is a common feature of strains isolated from patients with septicaemia, pyelonephritis and lower urinary tract infection.

Toxins
Pathogenecity of E. coli can also be attributed

to certain toxins which are mainly causing Diarrhoea

Enterotoxigenic E. coli Vero Toxin producing E. coli

Clinical Syndromes
Esch. coliis the most common cause of Acute, uncomplicated urinary tract infection
Hospital-associated urinary tract sepsis Neonatal meningitis Diarrhoea Septicaemia Sepsis in operation wounds Abscesses in a variety of organs.

As many as 80% ofEsch. colistrains that

cause neonatal meningitis and 40% of those isolated from infants with septicaemia but without meningitis express a K1 antigen. may be more virulent than those with other K antigens, as they share structural identity with host components.

Strains possessing the K1 or the K5 antigen

Urinary tract and septic infections

Urinary tract infection occurs more frequently

in women than in men because the shorter, wider, female urethra Most urinary tract infections are thought to be caused by organisms originating from the patient's own faecal flora Strains that cause urinary tract infection often originate from the gut of the patient, with infection occurring in an ascending manner. The ability ofEsch. colito infect the urinary tract may be associated with fimbriae that specifically mediate adherence to uroepithelial cells.

Diarrhoea
Esch. coliis normally carried in the gut as a

harmless commensal It may cause gastro-intestinal disease ranging in severity from mild, self-limiting diarrhoea to haemorrhagic colitis and the associated, potentially life-threatening,haemolytic uraemic syndrome. Strains fall into at least five groups, each associated with specific serotypes

1. Enteropathogenic Esch. coli(EPEC),

which cause infantile enteritis, especially in tropical countries 2. Enterotoxigenic Esch. coli(ETEC), which are responsible for community-acquired diarrhoeal disease in areas of poor sanitation and are the commonest cause of travellers' diarrhoea

3. Entero-invasive Esch. coli(EIEC),

which cause an illness resembling shigella dysentery in patients of all ages 4. Verocytotoxin-producing Esch. coli(VTEC), which cause symptoms ranging from mild, watery diarrhoea to haemorrhagic colitis and haemolytic uraemic syndrome

5. Entero-aggregative Esch. coli(EAggEC), which cause chronic

diarrhoeal disease in certain developing

Enteropathogenic Esch. coli (EPEC)

EPEC strains were originally identified as a cause

of diarrhoeal disease in infants Certain strains belonging to characteristically EPEC serogroups, such as O26 and O111 Colonization of the upper part of the small intestine occurs in infantile enteritis associated with EPEC. Electron microscopy of intestinal biopsy specimens has shown that the bacteria become intimately associated with the mucosal surface and are partially surrounded by cup-like projections ('pedestals') of the enterocyte surface. In areas of EPEC attachment the brush border microvilli are lost. Adhesion to the gut wall and the subsequent

Enterotoxigenic Esch. coli (ETEC)

ETEC produce a heat-stable enterotoxin or a heat-

labile enterotoxin, or both. In addition, they usually express fimbriae that are specific for the host animal species and that enable the organisms to adhere to the epithelium of the small intestine Infection is usually of brief duration, often beginning with the rapid onset of loose stools and accompanied by variable symptoms, including nausea, vomiting and abdominal cramps. Heat-labile enterotoxin (LT) is closely related to the toxin produced by strains ofVibrio cholerae. There are two main forms, termed LT-I and LT-II

Entero-invasive Esch. coli (EIEC)

EIEC, like shigella cause disease by invading

intestinal epithelium. Infection is by ingestion Only a small number of bacteria need to be swallowed as they are relatively resistant to gastric acid and bile, and pass readily into the large intestine where they multiply in the gut lumen. The bacteria pass through the overlying mucous layer, attach to the intestinal epithelial cells and are carried into the cell by endocytosis into an endocytic vacuole, which then lyses. After lysis of the vacuole the bacteria multiply within the epithelial cell and kill it. Spread to neighbouring cells leads to tissue destruction and consequent inflammation, which is

Verocytotoxigenic Esch. coli (VTEC)

Strains ofEsch. coliexpressing a protein

cytotoxic for Vero cells were discovered in 1977 Owing to the similarity in structure between Verocytotoxin and Shiga toxin expressed byShigella dysenteriae1, VTEC have also been termed Shiga toxin-producingEsch. coli. They are also sometimes referred to as enterohaemorrhagicEsch. coli. Human VTEC infection can be associated with a range of clinical symptoms from mild, non-bloody diarrhoea to the severe manifestations of haemolytic uraemic syndrome HUS

The biological properties, physical

characteristics and antigenicity of VT are very similar to those of Shiga toxin, produced by strains ofSh. dysenteriaetype 1 Like Shiga toxin, VT1 and VT2 comprise A and B subunits. VT1 and VT2, like Shiga toxin, are cytotoxic for Vero and HeLa cells Micro-angiopathy of the capillaries is a characteristic renal lesion in haemolytic uraemic syndrome, supporting the

Entero-aggregative Esch. coli (EAggEC)

Pathogenesis The mechanisms by which EAggEC cause diarrhoeal illness are poorly understood. Studies have failed to identify the infective dose The site of adhesion within the human host has not been determined. The characteristic pattern of adhesion to HEp2 cells may be a putative pathogenic mechanism. Strains of EAggEC may express fimbriae for

Lab Diagnosis
Specimen Urine Pus Stool CSF Morphology Gram Stain Culture Grow on

Blood Agar MacConkeys Agar

Colonial Morphology

Identification Motility Biochemical test Serology to detect type of O, H, K antigen Special test to detect toxin production
Biken Test Sereny Test CHO Test

Elisa Test RIA (to detect antibodies against toxins

Treatment and control

In the absence of acquired resistance,Esch.
ampicillin, cephalosporins, tetracyclines, quinolones aminoglycosides trimethoprim sulphonamides.

coliis susceptible to many antibacterial agents including

Many strains, however, have acquired plasmids

conferring resistance to one or more of these drugs Antimicrobial therapy should be guided by laboratory tests of sensitivity if possible.

Treatment and control ofEsch. colienteritis

General measures As with most diarrhoeal disease, the early administration of fluid and electrolytes is the most important single factor in preventing the death of the patient in severe infections. Despite the potentially serious consequences of VTEC infection, the use of antibiotics in this condition is controversial. The most effective means of preventing infection is to avoid exposure to the infecting agent. Contaminated food and water are probably the most important vehicles of ETEC infection in developing countries. VTEC infections are acquired most frequently from meat, unpasteurized milk and direct contact with

Vaccination No commercially available vaccine yet Antimicrobial prophylaxis A number of antimicrobial drugs reduce the incidence of diarrhoea in travellers to tropical areas. These include doxycycline, trimethoprim, norfloxacin and other fluoroquinolones