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

Abstract

ABSTRACTIn 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°C for up to 72 hours. Imipenem, meropenem, ertapenem, and doripenem discs from three brands, Oxoid, Becton Dickinson, and HiMedia, containing 10 μg 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 ofEscherichia coliwith different susceptibility profiles to carbapenems. The inhibition zone diameters were measured after 16-18 hours of incubation at 35±2°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°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 ABSTRACTHIGHLIGHTSIn remote areas, transportation conditions of AST discs may be challengingDiscs with four carbapenems from three brands were tested using six conditionsCarbapenem discs were generally stable for ASTIn unfavourable conditions, concentrations in the discs degraded over timeImipenem (Oxoid and Becton Dickinson) discs were the most stable