Effect of Bacteroides fragilis grown in the presence of clindamycin, metronidazole and fusidic acid on opsonization and killing of Escherichia coli.

Bactericidal action of human polymorphonuclear leucocytes on Escherichia coli in the presence of Bacteroides fragilis grown in subinhibitory concentrations of clindamycin, metronidazole and fusidic acid was studied. Bacteroides fragilis grown in the absence of drugs significantly inhibited the killing of Escherichia coli. Bacteroides fragilis grown in the presence of the drugs had a reduced inhibitory effect on the killing of Escherichia coli but this reduction was only significant for Bacteroides fragilis grown in 1/2 MIC of clindamycin. The phagocytosis of Bacteroides fragilis grown with and without clindamycin, as measured by killing, was the same. Complement consumption of Bacteroides fragilis grown with and without clindamycin did not differ. Clindamycin-treated Bacteroides fragilis fixed C3 to a significantly lower degree than did untreated bacteria. The chemiluminescence of Escherichia coli opsonized with serum preincubated with clindamycin-treated Bacteroides fragilis was significantly higher than with serum preincubated with untreated bacteria. These results suggested that in killing experiments of mixed Escherichia coli and Bacteroides fragilis, the mechanism underlying the reduced inhibitory capacity of clindamycin-exposed Bacteroides fragilis is related to greater availability of C3 in serum for opsonization of Escherichia coli.

Interactions between polymorphonuclear leucocytes, Bacteroides sp, and Escherichia coli: their role in the pathogenesis of mixed infection.

Five Bacteroides fragilis strains and five Bacteroides vulgatus strains were compared with regard to their ability to consume complement and to fix C3, their killing by polymorphonuclear leucocytes, and their ability to inhibit the bactericidal effect of serum and polymorphs on Escherichia coli strains. Complement consumption was positively related to C3 fixation, but no relation was observed between these variables and the killing of the anaerobes. Greatest inhibition of the killing of E coli by serum and polymorphs was achieved with the bacteroides strains that fixed most complement. The greater virulence of B fragilis in mixed infections with E coli was not reflected either by a greater ability to inhibit the killing of E coli or a greater resistance of the anaerobes themselves to the bactericidal effect of serum and polymorphs.

Haemagglutination by the Bacteroides fragilis group.

Adhesive properties of five species of Bacteroides were compared by direct haemagglutination with erythrocytes of different origin. Only strains of Bacteroides fragilis agglutinated erythrocytes and different patterns of haemagglutination were observed. None of eight carbohydrates tested inhibited haemagglutination. The activity was destroyed by heat and by periodate treatment, but not by three proteases tested.

Pathogenic synergy between Escherichia coli and Bacteroides fragilis or B. vulgatus in experimental infections: a non-specific phenomenon.

The virulence of Bacteroides fragilis and B. vulgatus for mice was compared in a skin-infection model. These strains were also tested for pathogenic synergy in mixed infections with Escherichia coli. Strains of B. fragilis were generally more virulent than strains of B. vulgatus and, with one exception, the effect of Bacteroides strains in mixed infections merely reflected their inherent virulence.

Pathogenic synergy between Escherichia coli and Bacteroides fragilis: studies in an experimental mouse model.

An animal model is described for quantitative evaluation of pathogenic synergy between Escherichia coli and Bacteroides fragilis in which adjuvants were not required for abscess formation. Two sets of strains of E. coli and B. fragilis isolated from human wound infections were tested. Pathogenic synergy was observed in only one of the two combinations and was dependent on properties of E. coli.

Killing of Escherichia coli by human polymorphonuclear leucocytes in the presence of Bacteroides fragilis.

The inhibitory effect of Bacteroides fragilis on the in vitro killing of Escherichia coli by polymorphonuclear leucocytes was studied with two pairs of E coli and B fragilis isolated from human wound infections. Both B fragilis strains behaved similarly: they inhibited the killing of one E coli strain, while the killing of the other E coli strain was not affected. The different behaviour of the two E coli strains depended on their need for fresh serum in the killing by the polymorphonuclear leucocytes. The inhibitory effect of the B fragilis strains could be completely accounted for by their effect on complement.

Polymorphonuclear leukocyte chemotaxis by mixed anaerobic and aerobic bacteria.

The induction of chemotactic activity of polymorphonuclear leukocytes (PMNL) by anaerobic and aerobic bacteria alone or in combination was evaluated. Washed cells as well as the supernate of Proteus mirabilis were chemotactic for leukocytes. The supernate of cultures of two strains of Bacteroides fragilis contained small amounts of chemotactic factors. No chemotactic factors were released from the non-fragilis Bacteroides strains. The supernates of cultures of anaerobic bacteria were capable of inhibiting chemotaxis of leukocytes to the chemotactic factors of P. mirabilis. P. mirabilis and two strains of B. fragilis generated chemotactic factors in serum but none of the other Bacteroides spp. tested were able to induce serum chemotactic factors.

Killing capacity of human polymorphonuclear leukocytes in aerobic and anaerobic conditions.

The killing by human polymorphonuclear leukocytes of several species of bacteria, some of which were catalase positive, was examined in vitro in aerobic and anaerobic conditions. When all conditions other than the oxygen tension were identical, killing after 30 min was slightly greater in aerobic than in anaerobic conditions. However, after 60 and 120 min the difference between aerobic and anaerobic killing was smaller, and killing was nearly complete for all strains tested. These results conflict with the common opinion that oxygen is essential for efficient killing. Minor differences in experimental conditions can greatly influence results, and may be responsible for the discrepancy between this study and some previous studies on this subject.

Pathogenic synergy: mixed intra-abdominal infections.

In this article we review our researches into the pathogenesis of mixed infections. These may conveniently be divided into in vitro and in vivo studies. In vitro we confirmed that interference with the killing of aerobes by polymorphonuclear leucocytes (PMN's) is a property of the Bacteroides strains tested and appears to depend on competition for opsonins i.e. complement factors. Further studies are in progress to define which complement factors and which bacterial structures are involved. The influence of B. fragilis on chemotaxis has also been studied. Our preliminary data suggest that B. fragilis is itself poorly chemotactic and reduces the chemoattractivity of Proteus mirabilis. This observation is surprising when we consider that abscess formation is the hall-mark of B. fragilis infections and needs clarification. In vivo we have developed a skin infection model in mice which is economical and gives reproducible and quantitative results. In this model we have demonstrated pathogenic synergy between Escherichia coli and B. fragilis. Further studies are planned to assess the role of complement and bacterial factors in this in vivo synergy.