Biomedical subjects
Publications and source records attributed to H Chart.
Hens were exposed to small-particle aerosols containing different concentrations of Salmonella enteritidis phage type 4. They developed a systemic infection and some birds were still excreting the organism in the faeces when killed 28 days after infection. S enteritidis was present for a similar period in a wide range of alimentary tract issues and in the ovary and oviduct.
A total of 192 samples of serum from 113 Sri Lankan patients with clinical dysentery was examined for antibodies of the IgM class to the lipopolysaccharides (LPSs) of Shigella dysenteriae-1 and Escherichia coli O157:H7. By means of ELISA and immunoblotting, 59 patients were found to have serum antibodies to the LPS of S. dysenteriae-1 only. Four samples from one patient were found to contain serum antibodies to the LPSs of both S. dysenteriae-1 and E. coli O157:H7. Antibodies to the LPS of S. dysenteriae-1 were also detected in 16 samples from 25 children, from Sri Lanka, with no previous history of dysentery; one of these children also had antibodies to the LPS of E. coli O157:H7. Analysis of 16 samples from apparently healthy children in the U.K. showed that only one serum contained antibodies to the LPS of S. dysenteriae-1. This patient had a history of recent travel to Pakistan. The isolation of S. dysenteriae-1 remains the preferred test for the diagnosis of bacillary dysentery. The use of serology as a means of providing evidence of infection with S. dysenteriae-1, however may prove to be a useful adjunct to cultural techniques but needs to be validated in an area where this organism is endemic.
Samples of serum from 125 patients with haemolytic uraemic syndrome, were screened for antibodies of the IgA and IgM classes to the lipopolysaccharide (LPS) of Escherichia coli O157:H7. By means of the techniques of ELISA and immunoblotting, 48 samples were shown to contain antibodies of the IgM class to E. coli O157 LPS, while 42 samples contained IgA antibodies to this O-antigen. Thirteen patients produced IgM but not IgA antibodies to the LPS of E. coli O157:H7. Of 42 patients with IgA antibodies to the E. coli O157 LPS, seven did not produce antibodies of the IgM class. From this study, screening patients' serum for antibodies to E. coli O157 LPS of the IgA class, in addition to those of the IgM class, a total of 55 patients were shown to be seropositive. This constituted an increase is in the sensitivity of obtaining evidence of infection by this organism by 12%.
Sera from ten patients with positive brucella serology were used to investigate antibody cross-reactions between the O-antigens of Escherichia coli O157 and Yersinia enterocolitica O9. SDS-PAGE profiles of lipopolysaccharide (LPS), purified from strains of E. coli O157 and Y. enterocolitica O9, were reacted with sera by immunoblotting. All ten sera contained antibodies which bound to the LPS of E. coli O157, and five of these sera also contained antibodies which bound to the LPS of Y. enterocolitica O9. Absorption studies using these five cross-reacting sera indicated the existence of at least three epitopes exposed on the O-antigens of E. coli O157 and Y. enterocolitica O9. One antigen binding site appeared to be exposed on the LPS of both organisms, while one epitope was exposed on the LPS of E. coli O157 only, and another on the LPS of Y. enterocolitica O9 only.
Commercially reared chickens were challenged with Salmonella enteritidis phage-type (PT) 4 by aerosol, or via the conjunctiva. Inhalation of 2.9 x 10(2) or 4.2 x 10(3) S. enteritidis resulted in the production of IgG antibodies to the lipopolysaccharide (LPS) of S. enteritidis PT4. When the aerosol inoculum was increased to 2.4 x 10(5) bacteria per bird the antibody produced were predominantly of the IgM-class. Chickens challenged with 10(3) S. enteritidis PT4 via the conjunctiva mounted only a poor immune response. Increasing the challenge dose to 10(8) S. enteritidis resulted in the production of high-titre serum antibodies of both the IgG and IgM classes. Results from this study suggest that aerosols containing small numbers of S. enteritidis PT4 might be responsible for intraflock infection of poultry.
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.
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.
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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.
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.
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.
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.
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Fifty-eight sera, from 29 chickens originating from two layer flocks known to be naturally infected with Salmonella enteritidis phage type (PT) 4, were examined for antibodies to S. enteritidis. Using the techniques of immunoblotting and ELISA, antibodies to the lipopolysaccharide (LPS) of S. enteritidis were detected in 43 of 58 sera. Antibodies were of the IgG class and bound to the S. enteritidis LPS antigen O = 12. Bacterial agglutination reactions using whole-cell preparations of S. enteritidis and S. pullorum, correlated with anti-LPS antibody reactions as detected by immunoblotting and ELISA. A rapid means of screening chicken sera for antibodies to the LPS of S. enteritidis as an indicator of infection is discussed.
Enterotoxigenic Escherichia coli (ETEC) of serotype O114:H21, which produced only heat-labile enterotoxin (LT), gave mannose-resistant hemagglutination (MRHA) with bovine erythrocytes. One strain, E20738A, was shown to possess fimbriae of approximately 7.5 nm diameter. On SDS-PAGE two possible fimbrial polypeptides of molecular masses 17.5 and 15.5 kDa were seen; the 17.5-kDa band was the most prominent. Loss of LT and MRHA together from strain E20738A was associated with loss of a 100-MDa plasmid. An absorbed anti-strain E20738A serum reacted specifically with the 17.5- and 15.5-kDa polypeptides and bound to the intact fimbriae. This antiserum reacted positively in an ELISA with LT-positive E. coli strains of serogroups O8, O15, O48, O114, and O146. The antiserum did not react with ETEC carrying known colonization factors. The term coli-surface-associated antigen (CS) 17 has been used to describe the fimbriae.