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Goessens, W. H. F.

Publications and source records attributed to Goessens, W. H. F..

2 recordsLinked to original sources

Co-occurrence of VIM-positive non-aeruginosa Pseudomonas spp. and Pseudomonas aeruginosa in the hospital: A detailed molecular comparison of blaVIM-2-containing plasmids from environmental and clinical isolates

BackgroundPseudomonas aeruginosa is a bacterial pathogen responsible for severe hospital-acquired infections, and is capable of forming persistent reservoirs in hospital sink drains, creating a transmission risk. In our hospital, we have cultured not only carbapenemase-producing, Verona Integron-encoded Metallo-beta-lactamase (VIM)-positive P. aeruginosa, but also VIM-positive non-aeruginosa Pseudomonas spp., from sink drains. Previously, a blaVIM-2-containing, conjugative plasmid conferring carbapenem resistance was found in a clinical P. aeruginosa isolate in our hospital. ObjectiveTo investigate if other Pseudomonas spp. from our hospital also carried a blaVIM-2-containing plasmid, genetically characterize these plasmids, and compare these plasmids to publicly-available plasmid sequences to identify their source. MethodsWhole-genome sequencing was used to sequence chromosomes and possible plasmids from VIM-positive non-aeruginosa Pseudomonas spp. and VIM-positive P. aeruginosa strains. Hybrid assemblies were generated to reconstruct plasmid sequences. All isolates were obtained from environmental sampling or clinical cultures during a prolonged VIM-positive P. aeruginosa outbreak in our hospital. ResultsAn identical blaVIM-2-containing plasmid was found in six non-aeruginosa Pseudomonas (P. carnis, P. oleovorans, and a novel species) and in three P. aeruginosa isolates. The previously-reported blaVIM-2-containing plasmid was found in three additional P. aeruginosa isolates. All P. aeruginosa belonged to high-risk clones ST111 or ST446. The two plasmids were derived from both clinical isolates and isolates from sink cultures, and were unique to our hospital when compared to other publicly-available plasmid sequences. ConclusionsDuring a VIM-positive P. aeruginosa outbreak in our hospital, two closely-related, blaVIM-2-containing plasmids co-occurred in multiple non-aeruginosa Pseudomonas spp. and in high-risk P. aeruginosa clones ST111 and ST446. Non-aeruginosa Pseudomonas spp. cultured from either patient or sink samples may be important sources of carbapenem resistance genes, and should not be overlooked during outbreak investigations. Moreover, plasmid analysis is essential to fully understand transmission routes in hospitals.

microbiology↗

Stability of four carbapenem antibiotics in discs used for antimicrobial susceptibility testing

In low-to middle-income countries, microbiological laboratories often use disc diffusion for antimicrobial susceptibility testing (AST). Reliable AST of carbapenem antibiotics is crucial for treatment decisions and surveillance purposes. Transport and storage conditions of materials used for AST are critical and may be challenging in some settings, where temperature cannot always be controlled. This study aimed to test the stability of four carbapenems in discs for AST under unfavourable conditions, i.e., at room temperature and 35{degrees}C for up to 72 hours. Imipenem, meropenem, ertapenem, and doripenem discs from three brands, Oxoid, Becton Dickinson, and HiMedia, containing 10 g of antibiotic were included. Discs were exposed to six unfavourable conditions and the recommended storage-condition as control. Subsequently, disc diffusion testing following the EUCAST guidelines was performed with four well-defined strains of Escherichia coli with different susceptibility profiles to carbapenems. The inhibition zone diameters were measured after 16-18 hours of incubation at 35{+/-}2{degrees}C. All experiments were executed in triplicate. In parallel, the carbapenems degradation was observed using a spectrophotometric method. Our study revealed that carbapenem discs were generally stable for AST although the concentration of most carbapenem antibiotics in discs decreased over time. Overall, imipenem (Oxoid and Becton Dickinson) discs were the most stable. Meropenem discs were less stable when exposed to 35{degrees}C than at room temperature. Concentrations of carbapenems in HiMedia discs were higher than those in Oxoid and Becton Dickinson. For carbapenem AST using disc diffusion in a rural area, we recommend using imipenem discs from Oxoid or Becton Dickinson. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=82 SRC="FIGDIR/small/599257v1_ufig1.gif" ALT="Figure 1"> View larger version (29K): org.highwire.dtl.DTLVardef@1ca4d5aorg.highwire.dtl.DTLVardef@9c54bborg.highwire.dtl.DTLVardef@17ee220org.highwire.dtl.DTLVardef@12fcfc0_HPS_FORMAT_FIGEXP M_FIG C_FIG HIGHLIGHTSO_LIIn remote areas, transportation conditions of AST discs may be challenging C_LIO_LIDiscs with four carbapenems from three brands were tested using six conditions C_LIO_LICarbapenem discs were generally stable for AST C_LIO_LIIn unfavourable conditions, concentrations in the discs degraded over time C_LIO_LIImipenem (Oxoid and Becton Dickinson) discs were the most stable C_LI

microbiology↗