- Volume 42, Issue 1, 1995

The association of Aeromonas spp. with travellers’ diarrhoea was studied among 978 Finnish tourists travelling to Morocco in winter (n = 398) and autumn (n = 580) in 1989. Fifty-five isolates from diarrhoeal patients with (n = 16) or without (n = 39) a recent travelling history in a developing country were also included. In Morocco, Aeromonas spp. were isolated from 8.7 % of patients with diarrhoea and from 1.4 % of non-diarrhoeal tourists (p < 0.001). Aeromonas spp. were found as the sole pathogen in 5.5 % of patients (p < 0.001). Diarrhoea with multiple pathogens, including Aeromonas spp., was found in 3.1% of patients. Species identification by phenotypic and genotypic methods indicated that A. veronii biotype sobria (hybridisation group HG 8/10) and A. caviae (HG 4) were the most common Aeromonas spp. associated with travellers’ diarrhoea. A. hydrophila (HG 1) and A. caviae (HG 4) were common in patients acquiring diarrhoea in Finland. Ribotyping of strains within a species showed that all strains had different ribotypes although the tourists were infected during the same trip. This study suggested that only certain Aeromonas spp. were commonly found in travellers’ diarrhoea. However, the causative role of those species is unclear.

Clinical isolates of Aeromonas were grown at eight different temperatures from 10°C to 40°C. Whole cell lysates were examined by SDS-PAGE and major temperature-dependent changes to both protein and lipopolysaccharide (LPS) profiles were identified. Cells grown at the higher temperatures (37°C and 40°C) produced abundantly a protein of c. 60 kDa which was not detected at the lower temperatures. Temperature-dependent expressions of other proteins were also noted but these were more variable among the isolates. An effect of temperature on expression of lipopolysaccharides was also noted in that some strains produced significantly less O-polysaccharides at the higher temperatures. After fractionation of cells, major differences in the expression of cell envelope and outer-membrane proteins between cells grown at low and high temperatures were noted although no unifying patterns could be discerned. Such growth temperature-induced changes in the cell envelope constituents have not been described previously for Aeromonas isolates from man.

Urinary isolates of cysteine-requiring Escherichia coli were found to be generally lacking in virulence factors commonly associated with uropathogenic strains. The proportion of auxotrophic strains showing type-1 fimbriation, haemolysin production, motility and sensitivity to normal human serum was significantly less than that of a comparable number of urinary isolates of prototrophic E. coli, although the proportion in both groups possessing K1 antigen was similar. Furthermore, the biotyping and serogrouping of these and other strains from systemic infections demonstrated a high degree of phenotypic diversity. This is further evidence that infection with these auxotrophs results from a combination of decreased host resistance and a physiological condition conducive to the random selection of these auxotrophs in vivo.

The production of enterotoxin A, B, C and D by 196 Staphylococcus aureus strains isolated from blood cultures and 95 strains from nasal carriers was investigated. Half of the bacteraemia strains were from patients who died with or because of their infection, the other half from patients who survived. The nasal strains were selected to match the bacteraemia strains regarding phage types. Overall, 30.6% of the bacteraemia strains and 40.0% of the nasal strains produced enterotoxins; enterotoxins B and C were the toxins produced most frequently in both groups. A similar incidence and pattern of enterotoxin production was found among the bacteraemia strains of S. aureus regardless of acquisition of the infection, the portal of entry, presence or absence of endocarditis and outcome of the infection. Thus, the concept that the enterotoxins play an important role in staphylococcal infections, apart from the diseases caused by the toxins per se such as food poisoning and toxic shock syndrome, cannot be substantiated by the results of the present study.

Several inbred strains of mice in closed breeding colonies were found to have spiral-shaped bacteria associated with active, chronic hepatitis. A new species of Helicobacter, H. hepaticus, was isolated from the infected livers of some strains of mice. Other strains of mice were colonised with H. hepaticus in the caecum and colon, but not the liver. Filter-sterilised supernatant fluid from five strains of H. hepaticus was tested in a mouse liver cell line (ATCC no. CCL 9.1) for cytotoxic activity. All strains produced a toxic factor causing morphological changes in the cells at dilutions up to 1 in 1000. Toxicity was observed after exposure to the supernatant fluid for 48–72 h. Other Helicobacter spp. that also produced the cytopathic effect (CPE) in the liver cell line were H. felis, H. acinonyx, H. pylori and one strain of H. mustelae. “Helicobacter rappini” and H. muridarum did not cause CPE in the liver cells. The soluble factor was stable at 4°C for up to 3 months. It was also stable at 56°C for 30 min, but was inactivated by boiling for 15 min. It was inactivated by incubation with trypsin. A partially purified preparation of the cytotoxin had a mol. wt of c. 100000 and did not have urease activity. The cytotoxin produced by H. hepaticus did not cause vacuole formation in HeLa cells.

Clostridium difficile is isolated from the intestinal tracts of > 50% of healthy infants. The mechanism by which intestinal colonisation of infants by toxigenic C. difficile is generally asymptomatic is unknown but may reflect the presence in human milk of neutralising activity against C. difficile toxin A. On this basis, the ability of human milk to inhibit the binding of toxin A to a purified hamster brush border membrane receptor was determined. Ten milk samples from healthy volunteers in various stages of lactation inhibited the binding of toxin A to the receptor by an average of 90 %. Heating and dialysis did not significantly alter the inhibitory activity of any of the milk samples. Human milk protected adult hamsters against a lethal challenge with toxin A but had no effect on the cytotoxic activity of the toxin. SDS-PAGE and ligand blot analyses showed that there were at least four distinct factors in human milk that specifically bound toxin A. Thiophilic adsorption chromatography was used to separate immunoglobulin from non-immunoglobulin components of human milk. IgA was the only immunoglobulin detected in human milk and > 90 % of this immunoglobulin was recovered after purification by thiophilic adsorption. Both the unbound non-immunoglobulin and bound immunoglobulin fractions of human milk inhibited the binding of toxin A to the purified receptor. These results suggest that human milk may be important in protecting infants against C. difficile-associated intestinal disease.