Interpreting Susceptibility Testing of Carbapenem Against Pseudomonas aeruginosa

Objectives: Carbapenems are among the most powerful anti pseudomonal agents. Since meropenem and doripenem were marketed, there are limited data regarding drug susceptibility testing by routine methods (disc diffusion and Etest) for them. The aim of our study was to compare in vitro activity of the imipenem, meropenem and doripenem against Pseudomonas aeruginosa. Methods: Three hundred and eleven P. aeruginosa strains isolated from respiratory specimens in 170 patients who developed ventilator-associated pneumonia in two intensive care units were collected over a period of 31 months. The susceptibility of all of these isolates to imipenem, meropenem and doripenem were determined by Etest and disc diffusion method. Results: Considering either all of the isolates or only the first isolates recovered per patient (311 and 170 respectively) the susceptibility rate for doripenem was higher than that for meropenem and imipenem. When MICs determined by Etest were converted into interpretative categories (S, I, R) using French (CA-SFM) guidelines, agreement was poor, especially for meropenem and doripenem. The percent of agreement with the disc diffusion method were 90.6% and 89.7% for imipenem, 80.5% and 82.6% for meropenem and 80.5% and 73.3% for doripenem, for the first isolates and all of the isolates, respectively. Errors were mostly minor errors, and the rate of errors was as high as 17.7% and 16.1% for meropenem and 17.7% and 25.7% for doripenem for the first isolates and all of the isolates, respectively Conclusion: The accuracy of disc diffusion using CA-SFM guidelines appears unsatisfactory for all the three carbapenems justifying the adaptation of new guidelines for P. aeruginosa and carbapenems

Dual Protective Functions of Helicobacter pylori Peptidoglycan Modifications Impact Early and Late Survival in the Host

Structural motifs inherent in bacterial peptidoglycan (PG) are important recognition domains for some efficient host responses to pathogenic bacterial infection, so PG modifications by the infecting bacterium can neutralize host responses. Helicobacter pylori contain two PG modification enzymes, an N-deacetylase (PgdA) and an O-acetyltransferase (PatAB), but some naturally occurring strains lack the latter. Here the lysozyme resistance and the survival in both macrophages and in lysozyme deficient mice were studied for various H. pylori strains. Resistance to lysozyme killing of H. pylori was conferred by PgdA in naturally-occurring strains that lacked PatA (e.g. B128); the lysozyme sensitivity for B128 pgdA mutant was at the same level as that of a double mutant (pgdA patA) version within parent strain X47. Both PgdA- and PatAmediated PG modifications are important to H. pylori survival within macrophages. Mouse studies connected the lysozyme and macrophage results to the in vivo condition, in which lysozyme effects are expected early and cytokine production later, pertaining to specific host recognition events. An increased host lysozyme killing effect associated with the double mutant strain (pgdA patA) was indicated from a reduced colonization phenotype (compared to wild type) after a week post-inoculation of lysozyme-positive mice; at this time point some important anti- H. pylori mouse cytokines were shown to be minimal. At 3 weeks post-inoculation, the levels of 3 cytokines (IL-10, TNF-a, and MIP-2) were significantly higher in sera of mice inoculated with the pgdApatA strain, and the pgdApatA strain showed a greatly attenuated ability of mouse colonization, in contrast to the wild type strain, indicating a significant role of PG modifications in persistent infection through mitigating host immune response. However, the double mutant survived better in lysozyme deficient (LysM-/-) mice, supporting the notion that PgdA- and PatA- mediated PG modifications in H. pylori protect bacteria from killing by host lysozyme.