- Volume 31, Issue 4, 1990
Volume 31, Issue 4, 1990
- Articles
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Mice vaccinated with a non-virulent, aromatic-dependent mutant of Salmonella choleraesuis die from challenge with its virulent parent but survive challenge with Salmonella typhimurium
More LessSummary. BALB/c mice given a live vaccine of an aroA mutant of Salmonella choleraesuis by intraperitoneal (i.p.) injection were not protected against i.p. challenge with its virulent parental strain but were protected against i.p. challenge with either of two virulent strains of Salmonella typhimurium (O [1],4,[5],12). Vaccination with a live vaccine of S. typhimurium aroA protected against challenge with S. typhimurium but not with S. choleraesuis. Intraperitoneal administration of either aroA strain evoked high levels of serum antibody against the homologous lipopolysacharide (LPS) as determined by an enzyme immunoassay. Sera from vaccinated mice also reacted with heterologous LPS but at dilutions at least seven-fold lower than homologous endpoint titres. The vaccination schedule employed with either live-vaccine strain conferred an equal degree of resistance to challenge with Listeria monocytogenes. After mixed infection of mice with equal numbers of virulent S. typhimurium and S. choleraesuis by the i.p. route, the former was isolated in numbers at least 50 000 times greater than the latter from the liver and spleen between days 1 and 5. When mice were vaccinated i.p. with S. choleraesuis aroA, L. monocytogenes or P. multocida before mixed infection, neither serotype showed more than a slight predominance in the liver and spleen during the same period. Thus, in relative terms, vaccination with S. choleraesuis aroA or inoculation with unrelated bacteria suppressed the growth of virulent S. typhimurium in mice but allowed virulent S. choleraesuis to multiply. These results clearly show that S. choleraesuis 381 can multiply to kill immunised BALB/c mice.
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Examination of strains belonging to enteropathogenic Escherichia coli serogroups for genes encoding EPEC adherence factor and Vero cytotoxins
More LessSummary. Four hundred and forty-nine strains isolated from patients with diarrhoea and belonging to 13 enteropathogenic Escherichia coli(EPEC) Oserogroups were tested with a DNA probe for the EPEC adherence factor (EAF). Positive results were obtained with only 36 strains; they belonged to 10 O serogroups and flagellar typing showed they were usually of the “classical” EPEC serotypes. Thirty-four of the 36 EAF-positive strains showed localised adhesion to HEp-2 cells. The two remaining strains, of serotypes O114:H2 and O127:H4, showed low level or no adhesion to HEp-2 cells. No colonies hybridising with the EAF probe were identified in cultures from 115 faecal specimens from healthy children. Sixteen of the 449 strains hybridised with one or both probes for the Vero cytotoxin genes VT1 and VT2; 15 of the 16 strains belonged to serogroups O26 and O128. None of the strains hybridised with both the EAF and VT gene probes. These studies show that the great majorityof strains belonging to EPEC O serogroups do not possess the EPEC adherence factor or carry VT genes.
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Plasmid and chromosomal related toxin polymorphism of Escherichia coli serogroup O138; plasmid transfer and co-integration with pRP4
More LessSummary. DNA-DNA hybridisation was used to examine 89 isolates of Escherichia coli serogroup O138 for toxin gene content and location. Sixty-six isolates encoded toxin genes; 29 were STpa STpb VT2, 24 were VT2, four were STpa STpb, three wereSTpb VT2, three were STpb VT2 LT and one was STpa. None were K88-, K99- or F41-positive. The VT toxin gene was chromosomally located in all VT+ isolates tested whereas STpa, STpb and LT were plasmid borne. In co-transfer experiments with pRP4 as the mobilising plasmid, VT2 genes were not transferred to recipient E. coli K12 whereas STpa, STpb and LT genes were. Two VT2-positive isolates inhibited pRP4 transfer to K12 by a factor of 106 and rare transconjugants harboured a cointegrate plasmid comprising pRP4 (54kb) and a cryptic host plasmid (145 kb). In a K12 background, the co-integrate had transfer and incompatibility properties of pRP4 whereas in both progenitor E. coli O138 backgrounds the co-integrate was transfer inhibited and in one case resolved to give original plasmids.
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Enzyme immunoassay (ELISA) for detection of Clostridium difficile toxin B in specimens of faeces
More LessSummary. Antisera against Clostridium difficile toxin B were prepared in sheep and rabbit and were used in indirect and sandwich enzyme-linked immunosorbent assays (ELISA) for the detection of toxin B. Polyvinyl chloride and polystyrene microtitration plates were tested as solid phases for the assay. Both assays had a lower limit of detection for toxin B of 1 ng/ml. They were used to detect the presence of toxin B in 210 human faecal specimens and also in the culture supernatant fluids of C. difficile strains isolated from the faecal samples. There was a close correlation between the results of sandwich ELISA and those of cytotoxicity tests and isolation of C. difficile. Our sandwich ELISA method seems to be useful as a presumptive test for detection of C. difficile toxin B.
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Surface-associated properties of Streptococcus milleri group strains and their potential relation to pathogenesis
More LessSummary. Thirty strains from the Streptococcus milleri (anginosus) group (SMG) obtained from various sources were tested for a range of characters that could be associated with pathogenicity and the results were compared with those for type strains of S. sanguis, S. mutans and S. pyogenes. The SMG strains were heterogeneous in all tests. Most (18) belonged to one of the Lancefield groups with group F predominating. Adherence of strains isolated from abscesses to buccal epithelial cells was greater than that of other strains (p = 0.033). Compared with strains of S. sanguis, SMG strains were generally not aggregated by human saliva. They differed from the type strain of S. pyogenes in their relative ability to bind fibrinogen and fibronectin; they were less effective in binding fibrinogen (0.33-4.28% cf. 22% for S. pyogenes) and generally more effective in binding fibronectin (0.49-12.37% cf. 0.95%). Strains isolated from infections were statistically better at binding fibronectin than other strains (p <0.001). The ability of strains to adhere to saliva-coated hydroxyapatite (SHA) varied 10-fold, from 0.16-16.35%. The amount of fibronectin bound by SMG strains correlated with their ability to adhere to SHA (p <0.001). The hydrophobicity of the strains, as measured in the hexadecane partition assay, ranged from 0.0% to 99.0%. Some strains carried both positive and negative cell-surface charges and some strains with a highly hydrophobic cell surface also possessed a relatively high cell-surface charge. A minority of strains possessed a net positive cell-surface charge. Neither hydrophobicity nor cell-surface charge was related to the capacity of strains to adhere to SHA. Strains of SMG co-aggregated weakly with strains of Veillonella parvula, V. dispar, Actinomyces viscosus and A. naeslundii.
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The incidence of anaerobes in the sputum of patients with cystic fibrosis
More LessSummary. The number of anaerobes in selected sputum samples from patients with cystic fibrosis (CF) was investigated. When cultured by a semi-quantitative method, 26 (23.85%) of 109 sputum specimens from 21 CF patients contained > 105 cfu of anaerobic organisms/ml. Nine (42.7%) of the 21 patients produced sputum containing such concentrations of anaerobes on at least one occasion. Anaerobes were isolated from repeated sputum specimens from five patients. The anaerobes most often isolated were Bacteroides disiens, pigmented Bacteroides spp. and anaerobic grampositive cocci. Anaerobes were isolated more often from sputum liquefied by sonication than from unliquefied sputum, suggesting that they were unlikely to be oropharyngeal contaminants.
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A longitudinal study of the cultivable subgingival anaerobic bacteria isolated from sheep during the development of broken mouth periodontitis
More LessSummary. In a longitudinal bacteriological study of the cultivable subgingival anaerobic flora isolated from developing broken mouth periodontitis in sheep, samples were taken from five sheep on each of three farms on seven occasions over a period of 2.5 years. Ten different bacterial genera were isolated regularly but with fluctuating frequencies. Bacteroides and Fusobacterium organisms accounted for nearly 70% of the isolates. The Bacteroides and Fusobacterium isolates studied in detail from one farm were identified to species level. The fusobacteria comprised F. nucleatum-like organisms (68.6%). F. necrophorum (29.6%) and F. naviforme (1.8%). The Bacteroides spp. were divided into 11 main groups and included black-pigmented species similar to B. asaccharolyticus and B. gingivalis. On the farm studied in detail, the sheep could be allocated to two groups according to progression of periodontal disease. Most of the B. gingivalis-like isolates were from sheep with actively progressing disease, indicating that this organism may play a role in periodontal destruction in sheep.
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