The serological relationship between Yersinia enterocolitica O9 and Escherichia coli O157 using sera from patients with yersiniosis and haemolytic uraemic syndrome.

Sera from patients with yersiniosis, shown to contain antibodies to Yersinia enterocolitica O9; and sera from patients with haemolytic uraemic syndrome (HUS) caused by Escherichia coli O157, were used to investigate serological cross-reactions between Y. enterocolitica O9 and E. coli O157. Lipopolysaccharide (LPS) was isolated from strains of Y. enterocolitica O9 and E. coli O157 and reacted with sera by immunoblotting and ELISA. Sera from patients with HUS contained antibodies to the LPS of E. coli O157 only; 80% of sera from patients with yersiniosis contained antibodies to the LPS of Y. enterocolitica O9 and E. coli O157. This one-way cross-reaction was also detected using hyperimmune rabbit antisera.

Screening for enteropathogenic Escherichia coli in infants with diarrhea by the fluorescent-actin staining test.

The attaching effacing (AE) adherence property is now recognized as an important virulence characteristic of enteropathogenic Escherichia coli (EPEC). The fluorescent-actin staining (FAS) test (S. Knutton, T. Baldwin, P. H. Williams, and A. S. McNeish, Infect. Immun. 57:1290-1298, 1989), which is diagnostic for the AE lesions produced by EPEC (and Vero cytotoxin-producing E. coli), has provided an additional tool with which to investigate this important class of enteric pathogens. In this study, we screened for the AE adherence property in two groups of E. coli isolated from infants with diarrhea by using the FAS test and compared the results with those from O:H serotyping, localized adhesion to HEp-2 cells (LA), and the EPEC adherence factor (EAF) probe. Only 16 of 41 (39%) E. coli strains previously diagnosed as EPEC by O antigen serogrouping were FAS test positive, and of these only 12 belonged to recognized EPEC O:H serotypes; 9 strains which did belong to EPEC O:H serotypes were FAS test negative. Of a second group of 297 untyped E. coli, 7 (2.3%) were FAS test positive, and of these only 2 belonged to EPEC serogroups; 5 belonged to serogroups not regarded as EPEC serogroups or were nontypeable. Of the 23 FAS-test-positive strains identified, 10 were EAF probe positive and showed good LA; 13 were EAF probe negative and showed a quantitatively distinctly poor LA. EAF-positive and EAF-negative strains, however, showed equally good adhesion to human small intestinal mucosa. None of the FAS-test-positive E. coli hybridized with probes for Vero toxins 1 or 2. We conclude that the FAS test is diagnostic not only for classical EPEC and Vero cytotoxin-producing E. coli but also for EPEC strains which are not currently being diagnosed because they belong to serotypes not generally regarded as EPEC serotypes.

Haemolytic uraemic syndromes in the British Isles, 1985-8: association with verocytotoxin producing Escherichia coli. Part 2: Microbiological aspects.

In a three year study of children under 16 years with haemolytic uraemic syndrome faecal samples were examined for the presence of Verocytotoxin producing Escherichia coli (VTEC) using DNA probes and for free neutralisable Verocytotoxin in a Vero cell assay with specific antisera. There was evidence of VTEC infection in 58 of 185 (31%) samples. A total of 53 VTEC was identified from patients with haemolytic uraemic syndrome. Thirty eight VTEC belonged to serotype O157:H7 or O157:H-, 34 produced VT2 only, and four strains produced both VT1 and VT2. The remaining 15 VTEC belonged to nine different O serogroups; three strains produced VT1, 10 produced VT2, and two were positive for VT1 and VT2. Three control groups of patients without haemolytic uraemic syndrome were also examined. There was evidence of VTEC infection in 8%, 6%, and 4% of specimens from individuals with bloody diarrhoea, those with diarrhoea only, and healthy controls respectively. VTEC from the bloody diarrhoeal and diarrhoeal controls were O157:H7 but those from the healty controls could not be O serogrouped. This study confirms the association of VTEC, and particularly strains of O157:H7, with haemolytic uraemic syndrome. Strains producing VT1, VT2, or both toxins were isolated, although over 94% of VTEC produced VT2 alone or together with VT1.

Extended serogrouping scheme for motile, mesophilic Aeromonas species.

A total of 1,255 strains of motile, mesophilic Aeromonas species isolated from clinical and environmental specimens in the United Kingdom and 258 strains isolated in Australia, Brazil, Peru, and the United States were examined by using antisera for serogroups O1 to O44 (R. Sakazaki and T. Shimada, Jpn. J. Med. Sci. 37:247-255, 1984) and for unpublished serogroup O45 (R. Sakazaki). The typeability rate for strains isolated in the United Kingdom was 35%; the strains isolated in other countries had typeability rates of between 14 and 43%. A total of 52 provisional new serogroups were identified, and the strains with unidentified O groups were examined by using antisera for these provisional new serogroups. The typeability rate for strains isolated in the United Kingdom was increased to 66% (70% of smooth strains). The typeability rates were 76% for A. hydrophila and 63% for both A. caviae and A. sobria. The 52 antisera for the provisional new serogroups increased the typeability rate for strains isolated outside the United Kingdom to between 43 and 68%. This extended serogrouping scheme would be of value in determining the importance of Aeromonas strains as human intestinal pathogens and in investigating the pathogenic mechanisms that may be involved in the production of diarrheal disease.

Adhesion to cells in culture and plasmid profiles of enteropathogenic Escherichia coli isolated from outbreaks and sporadic cases of infant diarrhoea.

Enteropathogenic strains of Escherichia coli (EPEC) that caused 10 outbreaks of infant diarrhoea in the U.K. between 1968 and 1986 were studied. All gave localised adherence (LA) to HEp-2 cells, HeLa cells and Intestine 407 cells in culture. All hybridised with the EPEC adherence factor (EAF) probe. The hybridising sequences were carried on plasmids ranging in size from 26 to 76 MDa. EPEC from sporadic cases of infant diarrhoea occurring between 1979 and 1986 that belonged to the same serotypes as the outbreak strains were also studied. All strains of serotypes O111ab.H2, O114.H2, O119.H6, O127.H6 and O142.H6 gave LA and were EAF-positive. In other serotypes, non-adhering strains or strains giving diffuse adherence were found also. In addition, strains of serotype O128.H2 which gave LA but did not hybridise with the EAF probe were identified. The strains isolated from sporadic cases of diarrhoea in the U.K. were similar, with respect to adhesion and hybridisation, to those isolated from sporadic cases of diarrhoea in developing countries.

A christening party outbreak of haemorrhagic colitis and haemolytic uraemic syndrome associated with Escherichia coli O 157.H7.

A point source outbreak of haemorrhagic colitis due to Escherichia coli O 157.H7 producing verocytotoxin (VT), took place following a christening party in Birmingham in June 1987. Twenty-six people were affected, six were admitted to hospital and one developed haemolytic uraemic syndrome: there were no deaths. VT + E. coli O 157.H7 was isolated from 13 (57%) of 23 faecal specimens from affected people and from 3 (9%) of 33 specimens from asymptomatic people. Free VT was detected in the faeces of one further asymptomatic person. Illness was associated with eating turkey-roll sandwiches (P less than 0.001) suggesting that cold meats might be an important source of infection.

Phage-typing of Vero-cytotoxin (VT) producing Escherichia coli O157 isolated in the United Kingdom.

Vero-cytotoxin (VT) producing Escherichia coli serogroup O157 have been isolated from patients with diarrhoea, haemorrhagic colitis (HC) and haemolytic uraemic syndrome (HUS). A phage-typing scheme developed in Canada has been used to type 155 VT+ E. coli O157 serogroup isolated from sporadic infections in the UK since 1983, and 48 strains from HC or HUS outbreaks. Twelve phage types were identified of which three, types 49, 51 and 52, have not been found in North America. All strains carried a 60 x 10(6) plasmid and most VT1+VT2+ strains also had a 5 x 10(6) plasmid coding for colicin D production. The majority of strains producing both VT1 and VT2 belonged to phage type 1, or the related types 4, 8 and 14. Most strains producing only VT2 belonged to types 2 or 49. Four outbreaks were included in the survey. Three had strains of a single phage type while strains from the fourth outbreak were more variable. The distribution of phage types throughout the UK showed no marked geographical variations.

Four new provisional serovars of Shigella.

Four bacterial strains are described that possess the biochemical characteristics of Shigella species but do not belong to any of the established Shigella serovars or to any previously described provisional serovar. One strain fermented mannitol, and it is proposed that this be the type strain for a new provisional serovar of Shigella boydii. The remaining strains did not ferment mannitol and belonged to three different serovars. These strains are proposed as type strains for three new provisional serovars of Shigella dysenteriae. All four strains were invasive in a HEp-2 cell tissue culture test, but only one was invasive in the guinea pig eye test and might therefore be expected to cause dysenterylike illness in humans. It is important that the designation of such strains remain provisional until other reference laboratories have had the opportunity to search for additional isolates and the possible pathogenicity of these strains for humans can be further assessed.

The use of sorbitol-MacConkey agar in conjunction with a specific antiserum for the detection of Vero cytotoxin-producing strains of Escherichia coli O 157.

Using DNA probes specific for the genes encoding Vero cytotoxins 1 and 2 in hybridization experiments on faecal samples, Vero cytotoxin-producing Escherichia coli (VTEC) of serogroup O 157 were detected in 21 of 63 cases of haemorrhagic colitis, 9 of 31 cases of non-bloody diarrhoea and 14 of 68 cases of haemolytic uraemic syndrome. Compared with these results sorbitol-MacConkey agar in conjunction with a specific O 157 antiserum gave a sensitivity of 62% in haemorrhagic colitis, 56% in non-bloody diarrhoea and 57% in haemolytic uraemic syndrome. The specificity of this method was 100% in all three groups. This demonstrates that sorbitol-MacConkey agar is a useful screening method for the detection of VTEC of serogroup O 157 when used in conjunction with a specific homologous antiserum. However, this method does not detect VTEC belonging to other serogroups and such strains were found, particularly in cases of haemolytic uraemic syndrome.

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.

Epidemic multiresistant Escherichia coli infection in West Lambeth Health District.

A year-long outbreak of multiresistant Escherichia coli K52 H1, predominantly serogroup O15, is reported from south east London. Most patients had urinary tract infections, some with septicaemia; but some cases of septicaemia were associated with pneumonia, meningitis, and endocarditis--unusual infections for E coli. 3 of these patients died. The organism was acquired in the community, and its source is still being investigated.

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.

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.

Outbreak of neonatal Citrobacter diversus meningitis in a suburban hospital.

Between February and June, 1983, four cases of Citrobacter diversus neonatal meningitis were identified at a suburban Baltimore hospital. One of the 4 infants died at age 13 months, 2 (both of whom had brain abscesses) have evidence of developmental delay and 1 appears to be normal after 33 months of follow-up. A review of microbiology records revealed that C. diversus had been present in the hospital nursery prior to identification of the first infant with meningitis, with isolation from infants born 7 months, 4 months and 4 days, respectively, before the first meningitis case. C. diversus was isolated from 21 infants born during the outbreak period and from hand or rectal cultures of 5 nursing personnel. All isolates were biotype E, with two distinct clusters of cases identified on the basis of plasmid profile and serotype. In a case-control study isolation of C. diversus was significantly associated with male sex, low birth weight and care by house pediatricians. The outbreak was controlled by stringent infection control measures and exclusion of personnel carriers. During the 24 months following the outbreak 3431 babies discharged from the nursery were cultured for C. diversus; 3 were colonized with the organism.

Molecular epidemiology of neonatal meningitis due to Citrobacter diversus: a study of isolates from hospitals in Maryland.

Six cases of neonatal meningitis due to Citrobacter diversus were diagnosed in three Baltimore (Maryland) hospitals between 1983 and 1985. Using plasmid profiles, biotypes, serotypes, and chromosomal restriction endonuclease digests as epidemiological markers, we studied 63 isolates of C. diversus (including four isolates from cerebrospinal fluid) from these and seven other hospitals in Maryland. Within two of the three hospitals with meningitis cases, the same strain of C. diversus was isolated from case infant(s), healthy neonates, and nursery personnel. In all three hospitals, C. diversus strains different from those implicated as a cause of meningitis were also isolated. Other than the meningitis-associated strains, 15 different strains of C. diversus were isolated from infants in the hospitals studied, with several distinct clusters of asymptomatic, colonized infants identified.