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Veterinary World, Vol.1, No.3, March 2008
Introduction
According to the Latin proverb, we are bornbetween the urine and faeces.Thus from birth, weacquire the faecal flora of our mothers.Over a century ago,Escherich described thebacteria that he isolated from the faeces of humanneonates as
Bacterium colicommune 
(Bettelheim,1986; Escherich,1988).He demonstrated that theorganisms now known as
Escherichia coli 
werepresent in the faeces and intestinal contents ofhumans and were considered as commensalorganisms.Most
E.coli 
strains are harmless commensals,however, some strains are pathogenic and causediarrhoeal disease.
E.coli 
strains that causediarrhoeal illness are categorized into specific groupsbased on virulence properties, mechanisms ofpathogenicity, clinical syndromes and distinct O: Hserogroups. These categories includeenteropathogenic
E.coli 
strains (ETEC),enteroinvasive
E.coli 
strains (EIEC), diffuse adhering
E.coli 
strains (DAEC), enteroaggregative
E.coli 
strains (EAggEC) and enterohaemorrhagic
E.coli 
strains (EHEC).EHEC were first identified as human pathogensin 1982, when
E.coli 
of serotype O 157:H7 wasassociated with two outbreaks of haemorrhagiccolitis. All EHEC produce factors cytotoxic to Africangreen monkey kidney (VERO) cells, which have beendescribed as verotoxins (VTs).E.coli O157:H7 andmany serotypes of
E.coli 
have subsequently shownto produce VTs; hence they have named VT-producing
E.coli 
(VTEC).
E.co l i 
O157:H7
The first confirmed isolation of
E.coli 
O157:H7in the United States was in 1975 from a Californiawoman with bloody diarrhoea.The first reportedisolation of
E.coli 
O157:H7 from cattle was from aless than 3 week old calf with colibacillosis inArgentina in 1977. The bacterium was first identified
E.co l i 
O157:H7
Treor Francis Fernandez30, Angalamman Nagar, Muthialpet,Puducherry-605003
as a human pathogen in 1982, when it wasassociated with two food borne outbreaks ofHemorrhagic colitis.Since then O157 VTEC have been identified inmany outbreaks and in sporadic cases of bloodydiarrhoea in North America and Great Britain and aclose association has been established betweenVTEC and haemorrhagic uremic syndrome (HUS).
Characteristics of E.coli O157:H7
Most strains of E.coli O157:H7 possess severalcharacteristics uncommon to most other E.coli.
(i) Acid tolerance:
Unlike most food bornepathogens,
E.coli 
O157:H7 uniquely tolerant to acidicenvironments. Acid tolerance is a complexphenomenon, both growth phase dependant andinducible.
E.coli 
cells in stationary phase of growthare substantially more acid tolerant than cells in theexponential phase. This increased tolerance isassociated with expression of genes regulated by therpoS sigma factor operon(Cheville
et al 
.1996;Rowbury,1998; Small
et al.
1994) examinedthree mechanisms of acid resistance, thatis,oxidative-arginine dependent and glutamatedependent, and found that all three contribute to themicroorganisms overall acid tolerance. Induction ofacid tolerance in
E.coli 
can enhance its survival inacidic foods (Cheville
et al 
, 1996; Layer
et al 
, 1995).
(ii) Antibiotic resistance:
Sufficient evidenceindicates that the bacterium is resistant to mostantibiotics.
(iii) Thermal inactivation:
Studies on the thermalsensitivity of
E.coli 
O157:H7 in ground beef haverevealed that the pathogen has no unusual resistanceto heat, with D values at 57.2°C,60°C,62.8°C and 64.3°C of 270,45,24 and 9.6 srespectively. Pasteurization of milk(72°C,16.2s) hasalso been determined to be an effective treatmentthat will kill more than 104
E.coli 
O157:H7 cells perml(D’Aoust et al,1988).Proper heating of foods ofanimal origin (63°C) is an important critical control
REVIEW
Veterinary World, Vol.1(3): 83-87083
 
Veterinary World, Vol.1, No.3, March 2008
point to ensure activation of
E.coli 
O157:H7.
(iv) Inability to grow well:
It is important to notethat many VTEC strains do not grow well at 44°C, ifat all, above 44°C.The minimum growth temperaturefor
E.coli 
O157:H7 under otherwise optimal conditionis approximately 8 to 10°C (Buchanan&Bagi, 1994).
(v) Inability to ferment sorbitol within 24 hours:
Most but not all O157 VTEC strains do not fermentsorbitol.
(vi) Inability to produce ß-glucoronidase:
Mostof the O157 VTEC strains will not hydrolyze 4-methylumbelliferyl-D-glucoronide.
(vii) Inability to produce gas and indole at 44°C:
Hence such methods would probably fail to detectVTEC.
(viii) Possession of an attaching and effacing(eae) gene:
This property contributes to VTEC toestablish its pathogenicity.
(ix) Carriage of a 60-MDa plasmid:
E.coli 
O157:H7isolates associated with human illness harbour aplasmid (pO157) of approximately 60 MD and thatcontains DNA sequences common to plasmidspresent in other serotypes of VTEC isolated frompatients with Haemorrhagic colitis. It washypothesized that the plasmid is believed to play arole in the pathogenicity of disease, but its functionis unclear.
(x)
Expression of an uncommon 5,000 to 8,000molecular weight outer membrane protein.
Reservoirs and sources of 
E.co l i 
O157:H7
Several reservoirs and sources of
E.coli 
O157:H7 have been identified. The association of
E.coli 
O157:H7 with undercooked ground beef andraw rice led to investigations of the role of cattle as areservoir of the pathogens. Several surveys of faecalshedding of
E.coli 
O157:H7 produced the followinggeneral observations.
l
Young animals tend to carry
E.coli 
O157:H7more frequently than adults(Zhao
et al 
.,1995)
l
Prevalence of faecal excretion variessubstantially among positive herds (Zhao
etal.
,1995).
l
E.coli 
O157:H7 levels in calf faeces rangefrom less than 10 2 CFU/g to 10 5 CFU/g(Zhao
et al.
,1995)
l
Faecal shedding of
E.coli 
O157:H7 frequentlyis intermittent and of short duration, i.e.several weeks to months (Brown
et al 
.,1997;Cray & Moon,1995)
l
More than one strain of
E.coli 
O157:H7 canbe isolated from faeces of the same animalor different animals within the same herd(Faith
et al 
., 1996; Ming
et al 
., 1995).
l
Calves have been experimentally infectedwith
E.coli 
O157:H7 (Brown et al.,1997; Crayand Moon,1995).The results revealed that,
1.
E.coli 
O157:H7 is not pathogenic to calves.Inoculation with 1010 CFU did not induce significantclinical disease.
2.
E.coli 
O157:H7 is continued to thegastrointestinal tract, with the fore stomachs (rumen,reticulum and omasum) and distal sites (distal ileum,proximal caecum, spiral colon and descending colon)being the principal sites of localization.
3.
E.coli 
O157:H7 did not form attaching andeffacing lesions and did not colonize mucosalsurfaces.The prevalence of
E.coli 
O157 in cattle has beenreported to range from 0.1 to 16%.The organism hasalso been isolated from the faeces of geese, sheep,horses, dogs, seagulls, goats and deer. The organismhas also been isolated from environmental sourcessuch as cattle, water troughs and soil.Sources of
E.coli 
O157:H7 for cattle have notbeen clearly identified. Possible sources includecontaminated feedstuffs or water,colonized animalsin herds,infected wildlife and humans or contaminatedfacilities and equipment surfaces from contact withfaeces.
Transmission of 
E.co l i 
O157:H7
Although a variety of foods have been implicatedin
E.coli 
O157:H7 associated illness, most outbreakshave been associated with consumption of raw orundercooked foods of bovine origin.
E.coli 
O157:H7infections also were associated with eating otherfoods, including vegetables, apple cider, cantaloupe,mayonnaise-containing salad dressing and salami.Contact of foods with
E.coli 
O157:H7 containing meator faeces (human or bovine) is a likely source ofcross-contamination. Person-to-person transmission(13.2%) and waterborne (4.4%) outbreaks have beendocumented.The mechanism of transmission in food chainis not fully understood, but contamination of meat fromintestinal contents at slaughter is probably animportant factor.
Detection of 
E.co l i 
O157:H7
(i) meat:
It is not easy to detect VTEC in raw meatsand foods where low levels of
E.coli 
may beswamped by high numbers of other bacteria. So far,there are no widely recommended methods forroutine food examination. Currently traditional
E.coli O157:H7
084
 
Veterinary World, Vol.1, No.3, March 2008
isolation methods for foods involve enrichment in aselective broth followed by plating on to sorbitol Macconkey agar with additives. This agar is only suitablefor 0157:H7 strains (most but not all 0157 VTECstrains do not ferment sorbitol).The composition ofthe enrichment broth and plating agar is important ifVTEC is to be isolated from contaminated materialsand several researches are going on determining theoptimum combination of selective agents.
(ii) Stool:
Methods used in medical laboratories todetect the organism from stools are more successful,probably the number of VTEC cells present in thestools of someone made ill by the organism isrelatively high in comparison to the background flora.For
E.coli 
O157 (but not all VTEC serotypes)commercial kits are available for isolation andidentification (ELISA methods) and for confirmationof suspect colonies (latex agglutination).Recipes forseveral effective broths and agars have beenpublished but there is no consensus yet on which isthe best. Another technique used to enhance isolationof VTEC from enrichment broths is the use ofcommercially available immunomagnetic beadscoated with specific O157 antiserum. Animmunoblotting technique is available for rapididentification of O157 colonies on agar plates.For epidemiological purpose/surveillance O157VTEC can be distinguished by phage typing (Ahmed
et al 
., 1987) which can be combined with typing ofthe VT genes to give added discrimination.Other methods such as plasmid profile analysis,pulsed field gel electrophoresis and multilocusenzyme electrophoresis may further differentiateO157 VTEC and can be applied to VTEC of otherserogroups. Restriction fragment lengthpolymorphism analysis of genomic DNA probed withphage ë or the DNA of a VT encoding phage havealso been used to differentiate O157 VTEC strains.Thus for surveillance, isolation of
E.coli 
O157 shouldbe attempted using readily available methods (e.g.:cefixime tellurite sorbitol Mac conkey Agar) andtesting of non-forming sorbitol colonies with 0157antiserum. Biochemical and serological confirmationtests may also be needed to exclude falsepositives.VTEC strains other than 0157 do not havebiochemical markers that assist in their identification.The general methods which rely on enrichmentfollowed by plating onto selective agar andbiochemical confirmation all at 37°C would be morelikely to isolate VTEC. Method developed usingcultural and rapid techniques are progressing fast.
Characteristics of the disease
Fortunately infection from
E.coli 
0157 isrelatively rare. Its principal symptom, diarrhoea, isalso a symptom of other gastrointestinal infections.This has meant that the relatively few
E.coli 
O157have led to be found from among many moreinfections with this routine symptom.
E.coli 
O157 infections are associated with arange of illness in humans, although a proportion maybe asymptomatic. Where symptoms do occur, theincubation period is 2 to 10 days, with most casesoccurring in 3 days.The range of clinical disease includes:
l
Mild diarrhoea, fever, abdominal pain,vomiting
l
Hemorrhagic colitis (HC), which consists ofinflammation of the large bowel, with severeblood.
l
Haemolytic Uraemic syndrome (HUS), acombination of anaemia, acute kidney failureand low platelet count which may beaccompanied by fever.
l
Thrombotic thrombocytopenic purport (TTP)characterized by fever, via skin and centralnervous system involvement, resulting fromaggregation of platelets in various organs.HUS largely affects children and TTP largelyaffects adults.TTP is a rare syndrome of
E.coli 
O157:H7 infection.
Immunity to
E.co l i 
O157:H7
An infected patient’s serological responseagainst surface epitopes of
E.coli 
O157:H7 can lastfrom weeks to months. It can be useful forepidemiological studies to determine the serologicalresponses of patients suspected of
E.coli 
O157:H7infection when stool cultures are negative for recoveryof
E.coli 
O157:H7. However, such studies are limitedby the fact that all patients who were
E.coli 
O157:H7culture positive have a demonstrable antibodyresponse. Oral inoculation of calves and steers with1010
E.coli 
O157:H7 induced prompt and sustainedincreases in serum antibodies to the O157 antigenicLPS and to a lesser extent to Stx 1(Johnson
et al 
.,1996).The serological responses, however, do notcorrelate with elimination of carriage by cattle orprotection of calves against reinfection by the samestrain. The ability of
E.coli 
O157:H7 to persist in andreinfect cattle that have a strong immune responseis likely to contribute to the introduction andpersistence of infection in herds.
E.coli O157:H7
085
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