Does bacteremia pose a direct threat to synthetic vascular grafts?

This study was undertaken to determine the significance of graft lumen exposure to blood-borne organisms in the development of graft infection. Three groups of dogs were studied. In group I (n = 20), the infrarenal aorta was dissected from surrounding tissue, divided, and reconstructed with a Dacron tube interposition graft. In group II (n = 9) the aorta was similarly isolated, but Dacron graft material was wrapped around the intact aorta. In group III (n = 13) the infrarenal aorta was isolated, but no graft material was placed. All dogs were given intravenous 1 X 10(7) Staphylococcus aureus at the completion of surgery. Group I grafts were harvested 8 hours, 1 day, or 21 days after bacterial challenge. Group II and III grafts were harvested 1 day or 21 days after infusion. At the time of harvest, selective cultures of the periaortic tissue (PAT), periaortic graft (PAG), and interposition graft lumen (GL) were taken. The overall infection rates were similar, with 17 of 20 (85%) dogs in group I, 6 of 9 (67%) in group II, and 11 of 13 (85%) in group III found to be culture positive. In group I, 3 of 3 dogs at 8 hours, 2 of 2 on day 1, and 12 of 15 on day 21 had positive PAT cultures. Only 4 of 15 on day 21 had positive GL cultures. In group II, 4 of 5 dogs on day 1 and 2 of 4 on day 21 had positive PAT and PAG cultures. In group III, 9 of 9 animals on day 1 and 2 of 4 on day 21 had positive PAT cultures. All aortic lumen cultures were negative in groups II and III. The difference between GL and PAT cultures was statistically significant in all groups (I, p = 0.01; II, p = 0.05; III, p = 0.01). Serial quantitative blood cultures revealed a mean bacterial load of 10.5 +/- 4.5 CFU/ml at 15 minutes postinfusion, which fell steadily until no bacteria were detected at 3.5 hours. Lymphangiography demonstrated periaortic pooling of lymph in the immediate postoperative period. These data suggest that the bacteremia in this model is transient and rapidly clears. Periaortic tissues quickly sequester bacteria, possibly because of lymphatic leakage. The GL appears to be secondarily infected.

The penicillin binding proteins of the genus Haemophilus.

We questioned whether the penicillin binding protein (PBP) profiles of representative strains from the 19 species varied within the genus Haemophilus and whether these profiles would be of taxonomic value. Seventeen of the 19 representative strains studied had distinct PBP profiles; only those of H. avium and H. paragallinarum were identical. The data support the inclusion of H. aegyptius in the genus as a species related to but separate from H. influenzae and could not exclude H. somnus, H. agni, and H. equigenitalis from the genus. Comparative PBP analysis within the genus Haemophilus may therefore be useful taxonomically.

Ampicillin resistance and penicillin-binding proteins of Haemophilus influenzae.

Penicillin-binding protein (PBP) alterations have been associated with non-beta-lactamase-mediated ampicillin resistance in Haemophilus influenzae. We evaluated the PBP profiles of several ampicillin-susceptible and -resistant clinical isolates of H. influenzae to determine how consistently the described alterations occurred, and to document the reproducibility of the PBP profiles for this species. The MIC of ampicillin ranged from 0.06 to 0.13 microgram ml-1 for the susceptible isolates at an inoculum of 100,000 c.f.u. when tested by broth dilution, and was 0.5 microgram ml-1 for all four isolates when tested by agar dilution. The MIC for the resistant isolates ranged from 4 to 8 micrograms ml-1 when tested by broth dilution, and from 1.5 to 16 micrograms ml-1 when tested by agar dilution. At least eight distinct PBPs with molecular masses ranging from 27 to 90 kDa were detected both in cell membrane preparations and whole cell (in vivo) binding assays done on cells in the exponential growth phase. PBP variability was evident both in the ampicillin-susceptible and -resistant isolates; however, much greater variability existed within the four resistant strains. The differences in PBP patterns included (1) electrophoretic mobility, (2) binding capacity for the antibiotic and (3) the presence of additional PBPs in two of the resistant isolates. However, decreased binding capacity was consistently demonstrated in PBP 5 (56 kDa) of all of the resistant isolates. Saturation curves with both penicillin and ampicillin indicated that PBP 5 had decreased affinity for the antibiotics.(ABSTRACT TRUNCATED AT 250 WORDS)

Susceptibility to cephalosporins of penicillin-susceptible and penicillin-resistant strains of Neisseria gonorrhoeae from Philadelphia.

Using agar dilution, we determined MICs of penicillin, cefoxitin, ceftriaxone, cefmetazole, tetracycline, and spectinomycin for 129 strains of Neisseria gonorrhoeae. All strains were susceptible to ceftriaxone (MIC range, less than or equal to 0.008 to 0.06 micrograms/ml) and spectinomycin (16 to 32 micrograms/ml). The MICs for 50, 90, and 100% of strains tested were 1.0, 2.0, and greater than 8.0 micrograms/ml; 0.12, 1.0, and greater than 8.0 micrograms/ml; 0.5, 1.0, and 2.0 micrograms/ml; and 1.0, 2.0, and greater than 8.0 micrograms/ml for cefmetazole, penicillin, cefoxitin, and tetracycline, respectively. Seven strains were beta-lactamase producers; eight were chromosomally resistant to penicillin. There was a log-linear relation for non-beta-lactamase-producing strains between the MICs of cefmetazole, cefoxitin, and tetracycline and the MIC of penicillin (Pearson r = 0.787, 0.544, and 0.358, respectively).

Genetic and phenotypic diversity among ampicillin-resistant, non-beta-lactamase-producing, nontypeable Haemophilus influenzae isolates.

Levels of genotypic and phenotypic diversity among 23 ampicillin-resistant, non-beta-lactamase-producing (Ampr NBLP) isolates of serologically nontypeable Haemophilus influenzae recovered from the respiratory tract were determined by multilocus enzyme electrophoresis, auxotroph testing in chemically defined media, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of penicillin-binding proteins (PBPs). Twenty distinctive multilocus enzyme genotypes were identified, among which the average level of genetic diversity per locus was equivalent to that in the species as a whole. Hence, a single, recent origin for Ampr NBLP strains is excluded. Of the growth factors tested, a requirement for methionine was significantly associated with the Ampr NBLP phenotype. In contrast to the relative homogeneity of the PBP profiles of the ampicillin-susceptible strains tested (8 PBPs detected), the PBP profiles of the Ampr NBLP strains exhibited marked heterogeneity (5 to 10 PBPs detected). Care should be taken in interpreting changes in PBP profiles and in associating these profiles with resistance for species such as H. influenzae that demonstrate variability.

Relative penicillin G resistance in Neisseria meningitidis and reduced affinity of penicillin-binding protein 3.

We examined clinical isolates of Neisseria meningitidis relatively resistant to penicillin G (mean MIC, 0.3 micrograms/ml; range, 0.1 to 0.7 micrograms/ml), which were isolated from blood and cerebrospinal fluid for resistance mechanisms, by using susceptible isolates (mean MIC, less than or equal to 0.06 micrograms/ml) for comparison. The resistant strains did not produce detectable beta-lactamase activity, otherwise modify penicillin G, or bind less total penicillin. Penicillin-binding protein (PBP) 3 of the six resistant isolates tested uniformly bound less penicillin G in comparison to the same PBP of four susceptible isolates. Reflecting the reduced binding affinity of PBP 3 of the two resistant strains tested, the amount of 3H-labeled penicillin G required for half-maximal binding was increased in comparison with that of PBP 3 of the two susceptible isolates. We conclude that the mechanism of resistance in these meningococci relatively resistant to penicillin G was decreased affinity of PBP 3.

Cefuroxime treatment failure of nontypable Haemophilus influenzae meningitis associated with alteration of penicillin-binding proteins.

A 10-year-old boy presented with nuchal rigidity and cerebrospinal fluid (CSF) leukocytosis initially and again on day 6 of intravenous cefuroxime therapy (200 mg/kg/day). Both CSF specimens yielded nontypable beta-lactamase-negative Haemophilus influenzae that were susceptible by disk tests but relatively resistant to cefuroxime (MIC, 8- to 16-fold greater than that of control isolates). To define the mechanism of resistance, the cefuroxime resistance marker was transformed to a susceptible H. influenzae recipient; inactivation and permeability of beta-lactam substrate were tested and the penicillin-binding protein (PBP) profiles were examined. Inactivation of beta-lactam substrate was not detected and reduced permeability was not found. However, reduced beta-lactam binding to PBPs 4 and 5 was observed; 18- to 27-fold more penicillin and 2.5-to 4-fold more cefuroxime was required to saturate or block 50% of the binding sites of these PBPs, respectively. Thus, reduced affinity of PBPs 4 and 5 for beta-lactam substrate appears to be the mechanism of cefuroxime resistance in this strain. The reduced affinity of these targets appears to have contributed to the bacteriologic and clinical failure in this patient.