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Biomedical subjects

S M Scotland

Publications and source records attributed to S M Scotland.

At least 55 records · Page 3Linked to original sources

Antibodies to Escherichia coli O157 in patients with haemorrhagic colitis and haemolytic uraemic syndrome.

Twenty four sera from patients with haemolytic uraemic syndrome or haemorrhagic colitis and healthy controls were examined for antibodies to the lipopolysaccharide (LPS) of Escherichia coli O157. Faecal specimens from these patients were also examined for Vero cytotoxin producing E coli (VTEC) by DNA probes, and for faecal Vero cytotoxin. Eight patients with faecal E coli O157:H7 gave a strong antibody response to O157 LPS, shown by immunoblotting and an enzyme linked immunosorbent assay. Six symptomatic patients without evidence of faecal VTEC also gave a strong antibody response to O157 LPS. Sera from the remaining five patients and five healthy controls did not contain antibodies to E coli O157. The results suggest that the testing of sera from patients with haemorrhagic colitis or haemolytic uraemic syndrome by ELISA or immunoblot would prove valuable in addition to the established procedures for detecting VTEC, using DNA probes and testing for faecal Vero cytotoxin.

Antibodies, Bacterial↗

First recognized community outbreak of haemorrhagic colitis due to verotoxin-producing Escherichia coli O 157.H7 in the UK.

The first recognized outbreak of haemorrhagic colitis due to Escherichia coli O 157.H7 in the United Kingdom affected at least 24 persons living in East Anglia over a 2-week period. The illnesses were characterized by severe abdominal pain and bloody diarrhoea of short duration. Eleven patients were admitted to hospital and there was one death. Patients were mainly adult women who had not eaten out of the home in the 2 weeks before onset. Unlike previously reported outbreaks hamburgers were not the vehicle of infection, and a case-control study suggested that handling vegetables, and particularly potatoes, was the important risk factor.

Adult↗

Vero cytotoxin production and presence of VT genes in Escherichia coli strains of animal origin.

Vero cytotoxin (VT) production has been studied in 34 Escherichia coli strains isolated from animals with enteric diseases. The strains were tested by DNA hybridization with probes for VT1 and VT2 sequences and also in toxin neutralization experiments with specific antisera. Twenty bovine strains, belonging to nine different O serogroups, produced VT1 or VT2 but not both toxins. In contrast, all 14 porcine strains of four different O serogroups produced VT2 only. Six of these porcine strains, belonging to serogroups O138, O139 and O141, were isolated from cases of oedema disease. In general, the porcine isolates produced toxin at a lower level than the bovine strains.

Animals↗

HEp-2 adhesion and the expression of a 94 kDa outer-membrane protein by strains of Escherichia coli belonging to enteropathogenic serogroups.

Sixty strains of Escherichia coli belonging to enteropathogenic serogroups (EPEC) were examined for the ability to adhere to HEp-2 cells, the possession of the genes encoding EPEC adherence factor (EAF) and the ability to express an outer-membrane protein (OMP) of 94 kDa thought to be involved in bacterial adhesion to eukaryotic cells. An absolute correlation was found between HEp-2 adhesion and the possession of the genes encoding EAF. An OMP of 94 kDa was observed in the SDS-PAGE profile of most adhesive strains. In some strains this protein was prone to proteolytic degradation. An antiserum raised to a HEp-2 adhesive strain of EPEC did not react with the 94 kDa OMP of all EPEC which were EAF-positive and HEp-2 adhesive, indicating some interstrain antigenic variation of this protein. Although this 94 kDa protein was surface-exposed, specific antibodies binding to the 94 kDa protein in situ in the outer membrane did not interfere with adhesion of EPEC to HEp-2 cells. Therefore, these studies question the value of this protein as a potential vaccine component.

Bacterial Adhesion↗

Vero cytotoxin-producing strains of Escherichia coli from children with haemolytic uraemic syndrome and their detection by specific DNA probes.

Faecal specimens from 66 children with haemolytic uraemic syndrome in the United Kingdom were examined for strains of Escherichia coli producing Vero cytotoxin (VT). Initially, conventional bacteriological methods were used to identify colonies of E. coli which were then tested for VT production. Subsequently, specific DNA probes for VT1 and VT2 were used in hybridisation tests to detect VT-producing E. coli (VTEC). VTEC strains were isolated from 19 cases and in 15 they belonged to serogroup O157. Fourteen of these O157 strains possessed the flagellar antigen H7 and one was non-motile. The VTEC strains from the remaining four cases belonged to serotypes O26:H11, O104:H2, O153:H25, and O163:H19 together with a rough VT+ strain with flagellar antigen H51. The O157 strains hybridised with either the VT2 probe or both VT1 and VT2 probes. The other VTEC strains hybridised with either the VT1 or VT2 probe. Confirmation of the production of VT1 and VT2 in vivo was obtained by the neutralisation of faecal VT with specific antisera raised against these two cytotoxins.

Animals↗

Toxins.

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Animals↗

Haemorrhagic colitis and Vero-cytotoxin-producing Escherichia coli in England and Wales.

Vero-cytotoxin-producing strains of Escherichia coli (VTEC) were identified by the use of DNA probes in 39% of faecal samples from patients with haemorrhagic colitis in England and Wales. The patients with VTEC were distributed widely and their ages ranged from 2.5 to 86 years (mean 41). 3 patients died, including a child of 2.5 years. 30 of the 32 VTEC strains belonged to serogroup O157. Plating on sorbitol agar for non-fermenters followed by agglutination with a specific O157 antiserum was a useful screening method for O157 VT+ strains. However, it was not as sensitive as the DNA probe technique and did not detect VTEC of other serogroups.

Adolescent↗

Properties of strains of Escherichia coli belonging to serogroup O157 with special reference to production of Vero cytotoxins VT1 and VT2.

Fifty-four strains of Escherichia coli belonging to serogroup O157 were examined for the production of Vero cytotoxins VT1 and VT2, and for other properties such as plasmid content, resistance to antimicrobial agents and colicin production. Twenty-six strains from cases of diarrhoea, haemorrhagic colitis and haemolytic uraemic syndrome in humans produced VT. By serum neutralization tests and hybridization with DNA probes for VT1 or VT2, three classes were recognized which produced either VT1 alone or VT2 alone or both VT1 and VT2. These strains were of H type 7 or non-motile. The strains producing VT were sensitive to all the antimicrobial agents tested, and all carried at least one plasmid which had a molecular weight of c. 60 X 10(6). Seven strains of porcine origin and 21 strains of human origin did not produce VT or hybridize with either DNA probe. None of these strains was of H type 7. Of the 21 human VT- strains, 17 were of extra-intestinal origin and 18 were of H type 45. Twenty-three of the 28 VT-strains were resistant to at least one antimicrobial agent.

Cytotoxins↗

Heterogeneity of Escherichia coli phages encoding Vero cytotoxins: comparison of cloned sequences determining VT1 and VT2 and development of specific gene probes.

Phages coding for production of Vero cytotoxins VT1 or VT2 in strains of Escherichia coli serotype O157.H7 or O157.H- were morphologically indistinguishable. Their genome size and restriction enzyme digests of the phage DNA were similar. These phages were clearly different in these respects from a VT1-encoding phage isolated from a strain of E. coli O26.H11 (H19). However the VT1 region cloned from the phage originating in the E. coli O157.H7 strain was identical to the VT1 region previously cloned from the phage carried by H19. Sequences encoding VT2 that were cloned from the phage in E. coli O157.H- have been mapped and the VT2 region identified by transposon insertion. The cloned regions coding for VT1 or VT2 production had no similarities in the presence of restriction enzyme sites over a distance of about 2 kb, and two VT1-specific probes spanning a region of about 1.4 kb did not hybridize under stringent conditions with cloned VT2 DNA. A 2 kb HincII fragment contained the VT2 genes but hybridized to VT1-encoding phages and recombinant plasmids via flanking phage DNA. A 0.85 kb AvaI-PstI fragment was a specific probe for VT2 sequences and did not hybridize under stringent conditions to phages or plasmid recombinants encoding VT1.

Bacterial Toxins↗

Enterotoxigenic strains of Escherichia coli O128 are not restricted to subgroup ac but also belong to subgroups ab and abc.

Ninety strains of Escherichia coli O128 isolated in many different countries were examined. The majority (77 strains) belonged to the antigenic subgroup O128ab, and 41 of these strains produced heat-stable enterotoxin (ST), heat-labile enterotoxin (LT), or both. Eight strains were of the antigenic subgroup O128ac; six produced ST only, and two were nontoxigenic. Five strains were of the antigenic subgroup O128abc; two produced ST and LT, and three were nontoxigenic. Ten of the strains studied produced Vero cytotoxin, and all belonged to the antigenic subgroup O128ab. It was concluded that, contrary to the report of Guth et al. (B. E. C. Guth, M. L. M. Silva, I. C. A. Scaletsky, M. R. F. Toledo, and L. R. Trabulsi, Infect. Immun. 47:338-340, 1985), enterotoxigenic strains of E. coli O128 are not restricted to subgroup ac but also belong to the most common subgroup, ab.

Antigens, Bacterial↗

An outbreak of food-borne enterotoxigenic Escherichia coli diarrhoea in England.

An outbreak of diarrhoea with abdominal pain occurred among members of the staff of a school and their guests after a social function at which a cold buffet was served. Sixty people attended the function and 43 subsequently completed questionnaires. Of these, 27 had diarrhoea. The median incubation period was 36 h and the range 12-66 h. Food history analysis showed an association between diarrhoea and eating curried turkey mayonnaise. Stool specimens from 13 of those who developed diarrhoea were examined: Escherichia coli 06.H16 (producing heat-stable and heat-labile enterotoxins) was found in nine specimens and E. coli 027.H20 (producing heat-stable enterotoxin) in 11 specimens. Eight patients had both strains and only one was negative for enterotoxigenic E. coli. Food samples were not available for examination. Enterotoxigenic E. coli should be considered as a possible cause in well-defined outbreaks of food-borne diarrhoea with abdominal pain.

Abdomen↗

Cloning of genes determining the production of vero cytotoxin by Escherichia coli.

Sequences encoding the production of a cytotoxin (VT) active on Vero cells were cloned in Escherichia coli K12 from a VT-determining phage that originated in E. coli strain H19 of serotype O26.H11. Subcloning resulted in the identification of a 2.5 kb fragment that still coded for VT production. Mutagenesis with transposon Tn1000 was used to map VT sequences and a 0.75 kb probe was developed. In colony hybridization tests with strains isolated from patients with haemolytic uraemic syndrome or diarrhoea, this probe derived from the H19 VT genes detected only some of the VT+ strains belonging to serogroup 0157. A VT+ strain, E32511, serotype 0157.H-, which was negative in colony hybridization was the source of another VT-determining phage from which VT sequences were cloned. Southern hybridization of the VT genes from E32511 with the H19 probe was negative under stringent conditions but there was weak homology under conditions of low stringency. These results indicate that there are differences in the VT genes of pathogenic E. coli.

Bacterial Toxins↗

Properties of wild-type strains of enterotoxigenic Escherichia coli which produce colonization factor antigen II, and belong to serogroups other than O6.

Enterotoxigenic strains of Escherichia coli, which belonged to serogroups other than O6 and produced colonization factor antigen II, usually produced only coli surface antigen 3 (CS3) and gave weak mannose-resistant haemagglutination of bovine erythrocytes. A non-autotransferring plasmid, NTP165, from a strain of E. coli O168. H16 coded for heat-stable enterotoxin, heat-labile enterotoxin and CS antigens. The CS antigens expressed after acquisition of plasmid NTP165 depended on the recipient strain: a biotype A strain of serotype O6. H16 expressed CS1 and CS3; a biotype C strain of serotype O6. H16 expressed CS2 and CS3; strain K12 and strain E19446 of serotype O139. H28 expressed only CS3. An exceptional wild-type strain, E24377, of serotype O139. H28 produced CS1 and CS3 when isolated; a variant of E24377 which had lost the plasmid coding for CS antigens produced both CS1 and CS3 after the introduction of NTP165.

Antigens, Bacterial↗