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B Rowe

Publications and source records attributed to B Rowe.

At least 163 records · Page 9Linked to original sources
PubMed1992

Strains of Escherichia coli O157:H8 from human diarrhoea belong to attaching and effacing class of E coli.

AIMS: To determine whether 17 Escherichia coli O157:H8 strains isolated from patients with diarrhoea in the United Kingdom were putative pathogens. METHODS: The strains had been isolated by the use of O157 antiserum, available for the detection of Vero cytotoxin (VT) producing strains of E coli O157 that are usually of flagellar (H) type 7, but may also be non-motile. The strains were examined for VT production, for their ability to adhere to HEp-2 cells, and for hybridisation with several DNA probes that recognise pathogenic properties of E coli. Their ability to ferment sorbitol and to produce beta-glucuronidase was also investigated, as these tests are used to discriminate VT positive O157 strains. RESULTS: The O157:H8 strains did not produce VT. All gave localised attachment to HEp-2 cells, associated with a positive fluorescence-actin staining test, and all hybridised with the E coli attaching and effacing (eae) probe. In addition to the difference in VT production, O157:H8 strains could be distinguished from VT positive O157 strains by their beta-glucuronidase activity, their failure to produce enterohaemolysin, and their lack of hybridisation with the CVD419 probe derived from a plasmid in an O157:H7 strain. CONCLUSIONS: The 0157:H8 strains had in vitro properties characteristic of the class of E coli that causes attaching and effacing lesions in epithelial intestinal cells. They may therefore be considered a putative cause of diarrhoea but their prevalence remains to be established. Several O157:H8 strains failed to ferment sorbitol in agar plates and therefore could be misidentified as VT positive O157 strains. Confirmatory tests for VT production are needed when O157 strains are isolated from faeces.

Bacterial Toxins↗
PubMed1991

Antibodies to lipopolysaccharide and outer membrane proteins of Salmonella enteritidis PT4 are not involved in protection from experimental infection.

BALB/c and Schofield mice were inoculated with formalin-killed bacteria prepared from strains of Salmonella enteritidis belonging to phage type (PT) 4 and carrying a 38 MDa plasmid and expressing long-chain lipopolysaccharide, or strains without a 38 MDa plasmid or lacking the ability to express lipopolysaccharide. Vaccinated mice were challenged with viable bacteria belonging to a virulent strain of S. enteritidis (PT4). Mice surviving this viable challenge were examined for a humoral antibody response to membrane antigens of S. enteritidis (PT4) that might relate to the possession of a given virulence property. BALB/c mice immunized with any of the test antigens were found to be immune to S. enteritidis (PT4), and this immunity was protective. Serum antibodies, of the IgG class, were detected to OmpA and a minor outer membrane protein (OMP) of 31 kDa. Schofield mice also raised IgG antibodies to these outer membrane proteins; however, non-immunized mice of this strain were resistant to infection. The virulence of S. enteritidis (PT4) was also tested using mice belonging to strains B10D2 (new), Biozzi (high), Biozzi (low), C3HeJ, B10ITYR and C57/L.

Animals↗
PubMed1991

Salmonella enteritidis PT4 infection in specific pathogen free hens: influence of infecting dose.

The period during which specific pathogen free hens, infected by direct introduction into the crop of either 10(3), 10(6) or 10(8) cells of Salmonella enteritidis PT4, excreted the organism in faeces was closely related to the size of the inoculum, with the birds excreting for mean periods of 3.4, 16.4 and 36.8 days, respectively. The production of either IgG or IgM was also dose related with the birds which received 10(3) cells having lower antibody levels than those in the other two groups. In contrast, there was no relationship between the contamination of egg contents and either antibody status, faecal excretion, or the dose administered.

Animals↗
PubMed1991

Cloning of regulator genes controlling fimbrial production by enterotoxigenic Escherichia coli.

Sequences regulating production of fimbriae were cloned from two enterotoxigenic Escherichia coli strains. One cloned region, from E. coli 0.25.H42, controlled expression of coli surface-associated (CS) antigen 4, whereas the function of the other, from E. coli 0167.H5, was unclear. Both regulators were related to the cfaD gene that controls expression of colonization factor antigen I (CFA/I) although low stringency conditions were required to show significant hybridization between cfaD and the regulatory fragment from E. coli 0167. The cloned regulatory genes promoted expression of CFA/I, CS1, CS2 and CS4 antigens but the levels of production in the presence of the 0167 regulator were lower than those promoted by the CS4 regulator or cfaD.

Antigens, Bacterial↗
PubMed1991

Serological identification of Escherichia coli O157:H7 infection in haemolytic uraemic syndrome.

To test the value of serological tests as an adjunct to bacteriological methods and toxin testing in haemolytic uraemic syndrome (HUS), 60 patients with the disorder were examined for evidence of faecal Escherichia coli producing verocytotoxin (VTEC), particularly of serogroup O157. They were also tested for serum antibodies reacting with the lipopolysaccharide of E coli O157 by means of an enzyme-linked immunosorbent assay (ELISA) and immunoblotting; for faecal VTEC by means of DNA probes hybridising with the genes encoding verocytotoxins VT1 and VT2; and for "free" faecal VT. Strains of E coli serotype O157:H7 were isolated from 9 patients, and faecal VT2 was detected in 3 of them. Strains of E coli of serotypes 05:H-, O55:H10, O105ac:H18, and O163:H19 were isolated from 4 patients, but faecal VT was not detected. Faecal VT2 was present in 1 patient from whom VTEC were not isolated. Antibodies to the lipopolysaccharide of E coli O157 were detected in serum samples from 44 patients. The 9 patients with faecal O157:H7 all had high titres of these antibodies, but serum samples from 16 healthy control children were negative. Serological testing of patients with HUS for antibodies to the lipopolysaccharide of E coli O157 provides evidence of infection with E coli O157 when faecal bacteria or VT cannot be detected.

Antibodies, Bacterial↗
PubMed1991

Purification of lipopolysaccharide from strains of Yersinia enterocolitica belonging to serogroups 03 and 09.

Lipopolysaccharide (LPS) was purified from strains of Yersinia enterocolitica belonging to serogroups 03 and 09, by three methods, and analysed by SDS-PAGE and silver staining for carbohydrate. SDS-PAGE of LPS prepared from whole-cells by digestion with proteinase-K, produced profiles containing high molecular mass LPS and a lower molecular mass region migrating as discrete bands. LPS prepared from strains belonging to serogroup 03, using a hot-phenol procedure alone was found to contain cellular proteins, and LPS prepared from strains of serogroup 09, by this method, did not contain high molecular mass carbohydrate. A novel method of preparing LPS by digesting bacterial outer membranes with proteinase-K prior to hot-phenol extraction produced protein-free LPS from strain of Y. enterocolitica 03 and high molecular mass LPS from strains belonging to serogroup 09.

Cell Membrane↗
PubMed1991

Applications of DNA probes for Vero cytotoxin-producing Escherichia coli.

Vero cytotoxin-producing Escherichia coli (VTEC) are now recognized as important aetiological agents in human disease. The symptoms of VTEC infection range from mild non-bloody diarrhoea to severe conditions such as haemorrhagic colitis (HC) and haemolytic uraemic syndrome (HUS). Two types of Vero cytotoxin (VT), VT1 and VT2, have been identified. The genes controlling production of VT1 and VT2 are phageencoded in several E. coli strains and DNA probes have been developed from cloned genes derived from these VT phages. Recently, synthetic oligonucleotide probes for VTEC have also been prepared and evaluated. The VT probes have been labelled radioactively and also non-radioactively with digoxigenin and biotin. Present applications of VT probes include detection of VTEC in faecal samples from patients with diarrhoea, HC or HUS and also the examination of different foods for the presence of VTEC. The proportion of VTEC in the faecal flora or foods may be low, often less than 1%. The use of DNA probes allows several hundred colonies from a sample to be examined and by this technique VT genes were detected in 30-40% of faecal specimens from patients with HC or HUS. Use of methods such as the polymerase chain reaction for amplification of the target genes combined with DNA probes should result in an increased sensitivity for the detection of VTEC.

Bacterial Toxins↗
PubMed1991

Non-radioactively labelled polynucleotide and oligonucleotide DNA probes, for selectively detecting Escherichia coli strains producing Vero cytotoxins VT1, VT2 and VT2 variant.

Vero cytotoxin producing Escherichia coli (VTEC) were detected in faecal specimens and bacterial isolates, using non-radioactively labelled polynucleotide and oligonucleotide DNA probes specific for Vero cytotoxin (VT) genes. VT1 and VT2 structural gene sequences, previously cloned and used for radioactive probes, were labelled with digoxigenin or biotin. Oligonucleotide gene sequences coding for the A subunit of VT1, VT2 and VT2 variant were labelled with digoxigenin. The VT1 and VT2 probes were specific for detecting VT1 and VT2 gene sequences and gave very similar results to those obtained using the radioactive label 35S as a standard. The VT2 variant probe hybridized only with the strains of porcine origin. For the range of isolates tested, there was little significant difference in specificity and sensitivity between the digoxigenin-labelled polynucleotide and oligonucleotide probes. The biotin system gave rise to more non-specific effects, particularly with some non-E. coli strains, and was therefore less reliable. All of the digoxigenin-labelled probes gave satisfactory results after several times re-use, which is of importance when considering cost.

Bacterial Toxins↗
PubMed1991

Instability of multiple drug resistance plasmids in Salmonella typhimurium isolated from poultry.

Plasmids in five strains of Salmonella typhimurium resistant to ampicillin, chloramphenicol, gentamicin, neomycin/kanamycin, streptomycin, sulphonamides, tetracyclines and trimethoprim (ACGKSSuTTm), CGKSSuTTm, ACSSuT or CSSuT which had been isolated from poultry in the first 3 months of 1989 have been characterized and compared with plasmids in two strains of R-types ACGKSSuTTm and ASSuTTm isolated from two patients later in the year. With the exception of the human isolate of R-type ASSuTTm, all strains carried two non-conjugative plasmids, one coding for SSu and belonging to incompatibility group Q, and a second coding for multiple resistance and belonging to the FIme incompatibility group. The human isolate of R-type ASSuTTm did not carry the IncQ SSu plasmid but like the poultry isolates, carried a non-conjugative FIme plasmid. Restriction endonuclease digestion with the enzymes EcoR I, Pst I and Hind III demonstrated that the FIme plasmids from strains of different R-types showed a high degree of homology but exhibited numerous fragment size polymorphisms. The restriction digest fingerprint of plasmids in the human isolate of R-type ACGKSSuTTm was indistinguishable from a poultry isolate of the same R-type. Analysis of segregants of one of the poultry isolates of R-type ACGKSSuTTm demonstrated that resistance determinants could be rapidly lost from the FIme plasmid to give rise to a number of R-types and fingerprint patterns. Loss of tetracycline resistance from this plasmid appeared to be correlated with the integration of other plasmid-mediated resistances into the bacterial chromosome. Evidence is presented for the rapid loss of antimicrobial resistance determinants from a multiple resistance plasmid of the FIme incompatibility group in response to withdrawal of antibiotic selective pressure.

Animals↗
PubMed1991

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.

Antibodies, Bacterial↗
PubMed1991

The influence of age on the response of SPF hens to infection with Salmonella enteritidis PT4.

When Specific Pathogen-Free hens were infected with Salmonella enteritidis PT4 by direct administration into the crop, the age of the bird at infection was found to have an effect on both pathogenesis and antibody response. Birds at 20 weeks of age showed no adverse signs and developed high titres of antibodies of the IgM class, while those which were 1 year old at infection developed relatively little antibody and had acute septicaemia, with 6 of 10 birds either dying or having to be humanely destroyed. The implication of these results for the control of salmonella infections in poultry is discussed.

Age Factors↗
PubMed1991

Surveys of human enterotoxigenic Escherichia coli from three different geographical areas for possible colonization factors.

Enterotoxigenic Escherichia coli (ETEC) from Burma, central Africa (Rwanda and Zaire) and Peru, were screened by enzyme-linked immunoassays for the colonization factor antigens (CFAs) and putative colonization factors (PCFs): CFA/I, CFA/II, which consists of three coli surface-associated (CS) antigens, CS1, CS2 and CS3, CFA/III, CFA/IV (CS4, CS5, CS6), CS7, PCFO9, PCFO159. H4, PCFO166, and CS17. The highest proportion of ETEC with identifiable colonization factors (71%) were found in the strains from Burma, which were mainly positive for CFA/I (38%), but strains producing CFA/II (4%), CFA/IV (11%), CS7 (10%), CS17 (4%), PCFO159, H4 (2%) and PCFO166 (2%) were also found. Sixty-nine percent of the ETEC from central Africa were positive for known colonization factors. While CFA/I positive strains were important (12%), a higher number of ETEC producing CFA/IV (33%) and CS17 (24%) were found. Fifty-two percent of the Peruvian strains produced identifiable colonization factors. The largest group of strains produced antigens of the CFA/IV complex (17%), while ETEC producing CFA/II (6%), CFA/III and CS6 (2%), CS7 (6%), PCFO9 (6%), PCFO166 (8%) and CS17 (7%) were also found. These surveys show that there is a considerable variation in the proportions and types of colonization factor found in different geographical areas. From 29 to 48% of the ETEC did not possess an identifiable colonization factor. These were particularly of the LT only producing type. These results have important implications for vaccine formulation.

Antigens, Bacterial↗
PubMed1991

Numbers of Salmonella enteritidis in the contents of naturally contaminated hens' eggs.

Over 5700 hens eggs from 15 flocks naturally infected with Salmonella enteritidis were examined individually for the presence of the organism in either egg contents or on shells. Thirty-two eggs (0.6%) were positive in the contents. In the majority, levels of contamination were low. Three eggs, however, were found to contain many thousands of cells. In eggs where it was possible to identify the site of contamination, the albumen was more frequently positive than the yolk. Storage at room temperature had no significant effect on the prevalence of salmonella-positive eggs but those held for more than 21 days were more likely (P less than 0.01) to be heavily contaminated. In batches of eggs where both shells and contents were examined, 1.1% were positive on the former site and 0.9% in the latter.

Animals↗
PubMed1991

Positive regulation of colonization factor antigen I (CFA/I) production by enterotoxigenic Escherichia coli producing the colonization factors CS5, CS6, CS7, CS17, PCFO9, PCFO159:H4 and PCFO166.

Entertoxigenic Escherichia coli (ETEC) strains of nineteen serogroups which produced colonization factors (coli-surface-associated antigens CS5, CS6, CS7 and CS17, colonization factor antigen CFA/III and putative colonization factors PCFO159:H4, PCFO166 and PCFO9) were tested for hybridization with a DNA probe containing the cfaD sequence that regulates expression of CFA/I. Strong colony hybridization, similar to that with the CFA/I-positive control strain H10407, occurred with ETEC strains of serogroups O27, O159 and O169 which produced CS6 antigen, and with all the strains which produced PCFO166 fimbriae. Weak colony hybridization, compared to the control strain, was found with ETEC producing CS5 fimbriae with CS6 antigen, CFA/III fimbriae with CS6 antigen, CS7 fimbriae or PCFO159:H4 fimbriae. CS6-antigen-positive strains of serogroups O79, O89 and O148 and all the CS17-antigen-positive and PCFO9-fimbriae-positive strains were negative in colony hybridization tests with the cfaD probe. Plasmid DNA of nine ETEC strains and their colonization-factor-negative derivatives was tested for hybridization with the cfaD probe and with ST and LT oligonucleotide probes. The sequences that hybridized with the cfaD probe were on the plasmids which coded for enterotoxin production. Fifteen strains were transformed with NTP513, a recombinant plasmid which contains the CFA/I region 1 fimbrial subunit operon but lacks a functional cfaD sequence, in order to determine whether DNA in any of these strains could substitute for the cfaD sequence in the regulation of production of CFA/I fimbriae.(ABSTRACT TRUNCATED AT 250 WORDS)

Agglutination Tests↗