Shiga Like Toxins Producing Escherichia coli

Noman Hafeez Khosa

M.Phil. Microbiology, Reg. No. F19-1520
University of Haripur, KPK, Pakistan

ABSTACT
Shiga toxins were named after the Kiyoshi Shiga who first described the dysentery due
to these types of toxins produced by Shigella dysentery. There are two major toxins
Shiga toxins1 and Shiga toxins2 (Stx1 and Stx2). These were initially mentioned similar
toxins as verocytotoxins produced by vero cells. Shiga like toxins of two types (SLT1
and SLT2) were then studied and found in some somatic (O) serogroups of Escherichia
coli strains O157:H7 as well as many other strains which were not found of much
importance initially. In many of the different cases, Enteropathogenic Escherichia coli
and Enterohaemorrhagic coli were also studied for the strain O157:H7 producing Shiga
toxins in vegetables. These toxins were being named as STEC when they were found
being produced by Escherichia coli. It was found that rather than the major Shiga toxins
producing Escherichia coli (STEC) strain O157:H7, many of the other strains of
serogroup (O) were also identified as Shiga toxins producing Escherichia coli (STEC)
including O104, O26, O111, O103 and O145 strains. Out the aforementioned strains
STEC O104:H4 was noticed as very common for Shiga toxins production and O26 and
O111 were having also been reported in many studies. The study designs used in
different studies were interviews, questionnaire, case-control studies, and cohort-
studies. However techniques used for detection of E.coli were, PFGE (pulse-field gel
electrophoresis), PCR (polymerase chain reaction), immunomagnetic separation (IMS)
procedures and culturing of E.coli. Owing to the multiple studies from different areas, we
found that leafy vegetables have been the major cause of carrying E.coli O157:H7
producing Shiga toxins. Many of them were reported due to unhygienic methodologies
of harvesting, transporting, storage and cooking.

Introduction Enteropathogenic Escherichia coli

Shiga toxin belongs to family of toxins
Stx1 and Stx2 which are expressed by a
gene eae which encode for adhesion
called intimin. Shiga like toxin (SLT) are
similar toxins to verocytotoxin, produced
by some serotypes of Escherichia coli
including strains O157:H7 and O104:H4.
(EPEC) is also only termed as E.coli
associated with somatic (O) serogroups
alleged as pathogen. One of the
cytotoxin for which the vero cells are
responsible is produced by
Enteropathogenic Escherichia coli. It
was suggested in previous studies that
Shiga like toxin production is similar
toxin as verotoxin.
Shiga toxin producing Escherichia coli
were reported as an emerging infectious
agent in respect of food borne diseases.
The infections or diseases reported in
respect of SLT 1 and SLT2 ranges from
mild diarrhea, hemorrhagic colitis (HC),
hemolytic uremic syndrome (HUS),
thrombotic thrombocytopenic purpura.
Hereby, we overview the Shiga like toxin
producing Escherichia coli in vegetables
which is going a serious health
challenge, previously studied mostly on
beef, meat and cattle farms and many of
other research areas.
Study Discussion
Shiga toxins are found in form of Stx1
and Stx2 which are expressed by the
genes that are part of genome of
lambdoid phages. Shiga toxin firstly
described in case of dysentery due to
Shigella dysentery and named after
‘Kiyoshi Shiga’ who described it.
Enteropathogenic Escherichia coli
(EPEC) also termed as E. coli
associated with somatic (O) serogroups
alleged as pathogens, albeit their
pathogenic mechanism doesn’t belongs
to enteroinvasion or to production of
heat stable or heat labile enterotoxin. A
cytotoxin which is detectable in Vero
cells is also produced by some EPEC
strains. The term Enterohaemorrhagic
Escherichia coli (EHEC) was also used
to study toxins producing strains those
causing hemorrhagic colitis (HC) and
hemolytic uremic syndrome (HUS).In
previous studies Shiga like toxins were
also noted similar toxin as verotoxin.
The major strain producing Shiga toxin
is O157:H7 but many of others have
also been studied in different periods of
times.
Shiga like toxin referred as SLT1 and
SLT2 are produced by Escherichia coli
and owing to which, they are also
abbreviated as STEC. STEC were
reported as an emerging infectious
agent in respect of food borne diseases.
The infections or diseases reported in
respect of SLT 1 and SLT2 ranges from
mild diarrhea, hemorrhagic colitis (HC),
hemolytic uremic syndrome (HUS),
thrombotic thrombocytopenic purpura.
For the identification of Shiga like
toxins1 (SLT1) and Shiga like toxins2
(SLT2) in vegetables, meat, cattle and
farm animals in Thailand, specific DNA
probes from were used. The cases afore
mentioned were literally reported in
children, adults and immuno-
compromised patients. Shiga toxin
producing E.coli causative agent for
human infection is found due to the
feckless ingestion of fecal matter, albeit
the unusual occurrence of infections due
to hemolytic-uremic syndrome with
prodromal diarrhea. The capability of
pathogenicity of STEC is occupied by
the many of agents of virulence; those
are Shiga toxins (Stx1 and Stx2), intimin
(eae), enterohaemolysin (Ehly) and
STEC auto agglutinating adhesin.
The study designs in different studies
conducted questionnaires, interviews,
case-control study and cohort study
along with analysis by using SAS
(statistical analysis software). The
laboratory techniques included were
PFGE (pulse field gel-electrophoresis),
PCR (polymerase chain reaction),
immunomagnetic separation (IMS)
procedures and cultivation of strains
with different culture media. It is easy
and economical to identify the
Escherichia coli O157:H7 in the
laboratory with the usage of sorbitol-
MacConkey (s-Mac) agar as E.coli
O157:H7 doesn’t ferment the sorbitol
after overnight incubation.
Studies have been carried out in respect
of Shige toxin producing Escherichia
coli in vegetables. An outbreak of
Escherichia coli O157:H7 in 1996,
among school children was reported in
Sakai city, Osaka, Japan. Schools of
those districts included in study had a
trend of serving lunch to school children
having same menu. This study was due
to consumption of white radish sprouts.
In this study hospitalized and non-
hospitalized children were studied
separately. Major symptoms in both
hospitalized and non-hospitalized
children were abdominal pain, diarrhea
and fever, albeit bloody diarrhea was
reported on an average of 33.5%. The
major reason was ignorance and
unavailability of hygienic conditions. The
unavailability of refrigeration or freezing
and other caring methods while
irrigation, harvesting, transporting and
storage. The probability of proliferation
of Escherichia coli O157:H7 was
confirmed in this study.
In another study for the prevalence of
EHEC O157:H7 from Nigeria, three
different types of vegetables, cabbage,
lettuce and tomatoes, were included.
The samples collected were 72 each of
cabbage, lettuce and tomatoes. This
study was actually carried out to
determine the presence of E.coli
O157:H7 in meat and vegetables sold in
Benin City, Nigeria. So hereby, it is
necessary to describe the sum of total
samples collected that was 432, and
216 samples were of vegetables, out of
these 432 samples. The Escherichia coli
found in cabbage, lettuce and tomatoes
were 48 (66.67%), 68(94.99%) and
64(88.8%) respectively. Escherichia coli
O157:H7 was not found in these
samples of vegetables.
During one of the study on meat and
vegetables, the samples that were
positive for Stx2 were also processed
for the identification of Shiga toxins
producing Escherichia coli strains O104,
O103, O111, O145, O26 and O157.
There were found no serogroup-
concerning genes in vegetables, albeit
found in meat samples. It was confirmed
in some of the studies that Shiga toxins
producing Escherichia coli (STEC) strain
O104:H4 is Stx2 positive negative for
Stx1 and intimin (‘eae gene).
According to According to Rangel et al.
(2), fresh produce resulted for 21% of
outbreaks and 34% of cases reported in
the United States between 1982 and
2002, and lettuce was the only most
regularly concerned product. The other
cases concerning freshcut lettuce, streptomycin, tetracycline, ciprofloxacin,
parsley, and spinach have been nalidixic acid, ampicillin, sulfisoxazole,
kanamycin, gentamicin, ceftriaxone and
reported since 2002. At that time of their
amoxicillin-clavulanic acid.
writing, an ongoing outbreak linked to
fresh-cut, packaged spinach has led to
199 confirmed infections resulting in 102
(51%) hospitalizations, 31 (16%) cases
of HUS, and 3 deaths (1.5%) (4). These
tragic vents clearly demonstrate that
leafy vegetables are currently a
significant vector for the transmission of
E. coli O157:H7 and point to the vital
necessitate for the development of
measures that will reduce the risk to
public health and restore confidence in a
key agricultural sector.
Human infection can result from direct
or in-direct contact with infected animals
(6, 8) or individuals (3, 7) and
contaminated surfaces

The elimination of pathogenicity of

salmonella and Escherichia coli
O157:H7 was studied by treating mung-
bean and radish sprouts. This study
resulted with the reduction or
inactivation of Escherichia coli O157:H7
to non-noticeable level. An adhesin
called intimin which is encoded by
genes ‘eae’ is found to be also
responsible for the attachment of STEC
strains to intestinal epithelial cells to
form lesions. Shiga toxins1 and Shiga
toxins2 (Stx1 and Stx2) were found
susceptible chloramphenicol, tri-
methoprim-sulfamethoxazole,
References 5. Maule, A. 2000. Survival of
verocyto-toxigenic Escherichia
1. Wells, J. G., B. R. Doris, and I. K.
coli O157:H7 in soil, water and on
Wachsmuth. 1983. Laboratory
surfaces. Symp. Ser. Soc. Appl.
investigation of hemorrhagic
Microbiol. 29:71–78.
colitis outbreak associated with a
6. Trevena, W. B., G. A. Willshaw,
rare Escherichia coli serotype. J.
T. Cheasty, G. Domingue, and C.
Clin. Microbiol. 18:512-520.
Wray. 1999. Transmission of
2. Rangel, J. M., P. H. Sparling, C.
Vero cytotoxin producing
Crowe, P. M. Griffin, and D. L.
Escherichia coli O157 infection
Swerdlow. 2005. Epidemiology of
from farm animals to humans in
Escherichia coli O157:H7
Cornwall and West Devon.
outbreaks, United States, 1982–
Commun. Dis. Public Health
2002. Emerg. Infect. Dis. 11:603–
2:263–268.
609
7. Valcour, J. E., P. Michel, S. A.
3. Boudailliez, B., P. Berquin, P.
McEwan, and J. B. March. 2002.
Mariani-Kurkdjian, D. D. Ilef, B.
Associations between indicators
Cuvelier, I. Capek, B. Tribout, E.
of livestock farming intensity and
Bingen, and C. Piussan. 2004.
incidence of human Shiga toxin–
Possible person-to-person
producing Escherichia coli
transmission of Escherichia coli
infection. Emerg. Infect. Dis.
O111–associated hemolytic
8:252–257.
uremic syndrome. Pediatr.
8. Warshawsky, B., I. Gutmanis, B.
Nephrol. 11:36–39.
Henry, J. Dow, J. Reffle, G.
4. Anonymous. 2006. Update on
Pollett, R. Ahmed, J. Aldom, D.
multi-state outbreak of E. coli
Alves, A. Chagla, B. Ciebin, F.
O157:H7 infections from fresh
Kolbe, F. Jamieson, and F.
spinach. Available at:
Rodgers. 2002. Outbreak of
http://www.cdc.gov/foodborne/e-
Escherichia coli O157:H7 related
colispinach/100606.htm.
to animal contact at a petting zoo.
Accessed 6 October 2006.
Can. J. Infect. Dis. 13:175–181.
9. Levine, M. M., J.-G. Xu, J. B. Consumption of Packaged
Kaper, H. Lior, V. Prado, B. Tall, Spinach, August–September
J. Nataro, H. Karch, and K. 2006: The Wisconsin
Wachsmuth. 1987. A DNA probe Investigation. Clinical Infectious
to identify enterohemorrhagic Diseases 2009; 48:1079–1086.
Escherichia coli of 0157:H7 and DOI: 10.1086/597399.
other serotypes that cause 13. Hideshi Michino, Kazuhiro Araki,
hemorrhagic colitis and hemolytic Shunsaku Minami, Satoshi
uremic syndrome. J. Infect. Dis. Takaya, Nobumichi Sakai, et al.
156:175-182. Massive Outbreak of Escherichia
10. Smith, H. R., B. Rowe, R. J. coli0157:H7 Infection in
Gross, N. K. Fry, and S. M. Schoolchildren in Sakai City,
Scotland. 1988. Haemorrhagic Japan, Associated with
colitis and Vero cytotoxin- Consumption of White Radish
producing Escherichia coli in Sprouts. American journal of
England and Wales. Lancet Epidemiology Vol.150, No. 8,
i:1062-1065. 1999; Pages 787-796.
11. A. So derstro m, P. O sterberg, A. 14. Marta-Louise Ackers, Barbara E.
Lindqvist, B. Jo nsson, et al. A Mahon, Ellen Leahy, Brant
Large Escherichia coli O157 Goode, Todd Damrow, et al. An
Outbreak in Sweden Associated Outbreak of Escherichia coli
with Locally Produced Lettuce. O157:H7 Infections Associated
Foodborne Pathogen and with Leaf Lettuce Consumption.
Disease. Vol. 5, No. 3, 2008, The Journal of Infectious
Pages 339-348 DOI: Diseases 1998; 177:1588–1593.
10.1089=fpd.2007.0065 15. M. L. Bari, Muhammad Imran al-
12. Arthur M. Wendel, Diep Hoang Haq, T. Kawasaki, M. Nakauma,
Johnson, Umid Sharapov, Juliana S. Todoriki, et al. Irradiation to Kill
Grant, et al. Multistate Outbreak Escherichia coli O157:H7 and
of Escherichia coli O157:H7 Salmonella on Ready-to-Eat
Infection Associated with Radish and Mung Bean Sprouts.
 ournal of Food Protection, Vol. Various Chemicals. Journal of
67, No. 10, 2004, Pages 2263– Food Protection, Vol. 62, No. 8,
2268. 1999, Pages 850–856.
16. Lia Bardasi, Roberta Taddei, 19. Markus Tzschoppe, Annett Martin
Lucia Nocera, Matteo Ricchi, and Lothar Beutin. A rapid
Giuseppe Merialdi. Shiga toxin- procedure for the detection and
producing Escherichia coli in isolation of enterohaemorrhagic
meat and vegetable products in Escherichia coli (EHEC)
Emilia Romagna Region, years serogroup O26, O103, O111,
2012-2013. Italian Journal of O118, O121, O145 and O157
Food Safety 2015; Vol. 4, No. strains and the aggregative
4511, Pages 33-35 DOI: EHEC O104:H4 strain from
10.4081/ijfs.2015.4511 ready-to-eat vegetables.
17. Azucena Mora, Santana L. León, International Journal of Food
Miguel Blanco, Jesús E. Blanco, Microbiology. 2012;152:19–30
Cecilia López, et al. Phage types, 20. Stephen A. Enabulele and Nduka
virulence genes and PFGE Uraih. Enterohaemorrhagic
profiles of Shiga toxin-producing Escherichia coli 0157:H7
Escherichia coli O157:H7 isolated Prevalence in meat and
from raw beef, soft cheese and vegetables sold in Benin City,
vegetables in Lima (Peru). Nigeria. African Journal of
International Journal of Food Microbiology Research Vol. 3,
Microbiology. 2007; 114:204-210. No. 5, 2009, Pages 276-279.
DOI:10.1016/j.ijfoodmicro.2006.0 21. Thomas G. Cleary, John J.
9.009. Mathewson, Elizabeth Faris, and
18. Peter J. Taormina and larry R. Larry K. Pickering. Shiga-Like
Beuchat. Behavior of Cytotoxin Production by
Enterohemorrhagic Escherichia Enteropathogenic Escherichia
coli O157:H7 on Alfalfa Sprouts coli Serogroups. Infection And
during the Sprouting Process as Immunity, Vol. 47, No. 1, 1985,
Influenced by Treatments with Pages 335-337.
22. M. Šafařĺková and I. Šafařĺk.
Immunomagnetic separation of
Escherichia coli O26, O111 and
O157 from vegetables. Letters in
Applied Microbiology 2001;
33:36-39.
23. Phillip I. Tarr and Marguerite A.
Neill. Perspective: The Problem
of Non-0157:H7 Shiga Toxin
(Verocytotoxin) - Producing
Escherichia coli. The Journal of
Infectious Diseases 1996;
174:1136-1139.
24. Orasa Suthienkul, J. Edward
Brown, Jitvimol Seriwatana,
Sanya Tienthongdee, Suvong
Sastravaha, and Peter
Echeverria. Shiga-Like-Toxin-
Producing Escherichia coli in
Retail Meats and Cattle in
Thailand. ASM: Applied And
Environmental Microbiology Vol.
56, No. 1, 1990, Pages 1135-
1139.
25. Phillip I Tarr, Carrie A Gordon,
Wayne L Chandler. Shiga-toxin-
producing Escherichia coli and
haemolytic uraemic syndrome.
Lancet 2005; 365: 1073–86.