- Volume 34, Issue 6, 1991

Twenty-four Haemophilus influenzae type b strains from 836 children and young adults in an open population were subtyped by outer-membrane-protein (OMP) analysis on sodium dodecyl sulphate-polyacrylamide gels, lipopolysaccharide serotyping and biotyping. The results were compared with those obtained with H. influenzae type b strains from 97 patients with meningitis in the same city (Amsterdam). OMP subtype 1 was significantly more common among the CSF isolates than in carrier strains (82% vs 50%; p≪0.002). The other OMP subtypes found among carriers were rarely isolated from patients. The lipopolysaccharide serotype and biotype distribution did not differ between the two groups. The combination of OMP subtype 1, lipopolysaccharide 1, biotype I was much more common in isolates from patients than in those from carriers (71% vs 42%; p≪0.01). The data suggest that various H. influenzae type b subtypes are less virulent than those commonly isolated from invasive infections.

The ability of various haem-and non-haem-iron-containing compounds to support the growth of iron-limited cultures of Haemophilus ducreyi was assessed in a plate bioassay. Only haemin or the haem-containing proteins, bovine haemoglobin, human haemoglobin and bovine catalase, but not equine cytochrome C1 1 1, were capable of serving as the sole exogenous iron source. Complexes of haptoglobin-haemoglobin and haem-serum albumin retained the ability to function as iron substrates. In contrast, no growth was observed with FeCI3, human lactoferrin and human transferrin. Siderophore production was not detected with a universal chemical assay. Outer-membrane-protein profiles derived from iron-starved cultures revealed four iron-regulated polypeptides of 65, 50, 40.5 and 40.5 Kda. These results indicate that haem can supply the requisite iron for growth of H. ducreyi.

Human lactoferrin (HLf) is an iron-binding protein and a host-defence component at the mucosal surface. Recently, a specific receptor for HLf has been identified on a strain of Staphylococcus aureus associated with toxic shock syndrome. We have looked for the occurrence of 125I-HLf binding among 489 strains of S. aureus isolated from various clinical sources. HLf binding was common among S. aureus strains associated with furunculosis (94.3%), toxic shock syndrome (94.3%) endocarditis (83.3%) and septicaemia (82.8%) and other (nasal, vaginal or ocular) infections (96.1%) with a mean binding (in fmol) of 29.1, 21.9, 16.9, 22.2 and 29.2 respectively; the differences between mean HLf binding values of 29.1–29.2, 21.9–22.2 and 16.9 were significant. Furunculosis-associated (low-invasive or localised) isolates were high-to-moderate binders of HLf; 50% gave positive results at a threshold of≫31 fmol of 125I-HLf bound. In contrast, endocarditis-associated (high-invasive or systemic) isolates demonstrated low binding and did not bind 125I-HLf at the above threshold level. S. aureus recognised human or bovine Lf. However, bound125I-HLf was more effectively inhibited in a dose-dependent manner by unlabelled bovine Lf than by homologous HLf. Binding of 125I-HLf to staphylococci was optimal with organisms grown in agar compared with those from broth cultures. The binding capacity of S. aureus was abolished when strains were grown on carbohydrate- and salt-rich agar media. HLf-binding ability of S. aureus did not correlate with fibronectin, fibrinogen, immunoglobulin G or laminin binding.

Trimethoprim was more potent than zidovudine as an inducer of the SOS response in Escherichia coli. The level of induction by each compound initially increased with rising drug concentration and then fell; this effect was less marked with zidovudine than with trimethoprim. The SOS response did not appear to be involved in the inhibition of bacterial multiplication as the MICs of trimethoprim or zidovudine for recA430 and lexA3 mutants, which are unable to induce the SOS response, were identical to the MICs for the parent strains. However, the bactericidal activity of each compound against strains deficient in the SOS response was reduced. This suggests that induction of the DNA repair system contributes to the bactericidal activity of the drugs.

Ninety-four clinical isolates of Pasteurella multocida of human origin were tested for dermonecrotic toxin (DNT) production by three methods: dermonecrotic test in guineapigs, Vero cell culture cytotoxicity and ELISA. The strains were isolated from patients living in a rural area with widespread intensive pig breeding. Six strains were found to be toxigenic by the three tests. A major protein band of Mr 145 Kda corresponding to DNT on immunoblots was demonstrated in extracts from these strains. All were isolated from respiratory tract (diseases 5, healthy carriage 1). The difference between isolates from the respiratory tract and isolates from wounds inflicted by pets was statistically significant with regard to DNT production (p≪0.02). A possible role of the toxin in pulmonary diseases caused by P. multocida has yet to be established.

Conventional biochemical and antibiotic sensitivity tests were used to allocate 87 clinical isolates of anaerobic gram-positive cocci to currently recognised species, in comparison with type and other authentic reference strains. Whole-cell protein electrophoresis was then performed with extracts of each strain. Allowing for difficulties of standardisation, it was possible to allocate most of the organisms to species-related groups on the basis of protein patterns. Organisms identified conventionally as Peptostreptococcus anaerobius and P. micros formed homogeneous groups by protein electrophoresis. There was evidence for heterogeneity amongst strains identified as P. asaccharolyticus (two groups, including P. indolicus), P. prevotii and P. magnus. However, aberrant P. prevotii strains were allocated to the P. asaccharolyticus groups, leaving a homogeneous P. prevotii group, and if P. variabilis were re-instated as a species, the remaining P. magnus strains could be divided into two groups. Of the anaerobic gram-positive cocci in the National Collection of Type Cultures deposited by Hare, Group IV is P. magnus, Group IX is P. micros and Groups I, III and VIII appear to be related to the butyrate-producing species P. asaccharolyticus and P. prevotii, but are strongly saccharolytic.

The kinetics of adherence of single isolates of Staphylococcus aureus, S. epidermidis, Pseudomonas aeruginosa and Escherichia coli to catheters made of polyvinyl chloride (PVC), Teflon®, siliconised latex, polyurethane and Vialon® was evaluated by a radiometric assay. Radiolabelled bacteria (108 cfu/ml) were incubated in vials containing 1-cm lengths of catheter for up to 3 days. The peak of maximal adherence to each biomaterial was reached after 24 h for P. aeruginosa and after 72 h for the other strains. Bacterial adherence to PVC and siliconised latex was significantly higher (2–6 times; p≪0.05) than to the other biomaterials for all the strains. The lowest values of adherence were observed with polyurethane and Vialon® for the staphylococci but with Teflon® for E. coli and P. aeruginosa. Bacterial viability and growth was evaluated in eluates obtained from incubation of segments of each catheter in buffer for 24 h. None of the eluates affected the viability of the staphylococci. However, all of them, significantly increased the growth of E. coli and P. aeruginosa with the exception of the eluate from siliconised latex, in which the inoculum count was reduced to an undetectable level for E. coli. We conclude that bacterial adherence to catheters may depend in part on the nature of the biomaterial and that certain substances eluted from the catheters may affect the viability and growth of different micro-organisms.

The virulence of transparent (Tr) and opaque (Op) colony types of Neisseria gonorrhoeae in the genital tract of female mice was evaluated at two stages of oestrous. Isogenic pairs of Tr and Op variants were isolated from N. gonorrhoeae strain 57–120. Both variants exhibited a T2 morphology, but only the Op variant possessed protein II (P. II) in outer-membrane fractions. When administered by intravaginal inoculation Op gonococci were highly infective only for mice in late pro-oestrous, whereas Tr gonococci were virulent for mice at both late pro-oestrous and dioestrous. Gonococci recovered from the uterus were of both Tr and Op phenotypes in equal proportions when mice were infected at dioestrous with Tr cells. In contrast, ≫90% of recovered colonies were of Op phenotype when mice were infected at late pro-oestrous with either Op or Tr cells. These results indicate that the virulence of gonococci for the genital tract of female mice differs from that for the chicken embryo. Furthermore, gonococcal survival in the female genital tract might be attributable to phase variation from Tr to Op phenotypes.

Eighty-six clinical isolates of Aeromonas hydrophila were studied for their ability to produce four exotoxins: a haemolysin active against rabbit erythrocytes, cytotoxin and enterotoxin detectable with Vero cell cultures, and the cholera toxin-like factor detected by an enzyme-linked immunosorbent assay. At least one exotoxin was produced by 80% of enteric and 96% of non-enteric isolates. The exotoxin profiles of non-enteric isolates were more restricted than those of enteric isolates, with haemolysin and cytotoxin producers preponderant. Although haemolysin and cytotoxin were produced by isolates from all sources, the enterotoxin and cholera toxin-like factor were more common amongst enteric isolates. The production of haemolysin and cytotoxin were closely related but the association between the enterotoxin and the cholera toxin-like factor was not significant.