A longitudinal study of antimicrobial resistant faecal bacteria in sediments collected from a hospital wastewater system

Abstract

Objective: The objective with this study was to determine and follow antimicrobial resistance in faecal bacteria over time in hospital wastewater pipe sediment. A further aim was to determine bacterial growth rates of sensitive, intermediate and resistant intestinal enterococci in different ciprofloxacin concentrations as a measure of bacterial fitness.Methods: A system enabling the collection of settled particles over time was installed at Kalmar County Hospital. Samples were collected bi-monthly for a 14-month period. Coliform bacteria and enterococci were isolated from the sediment with standard methods and investigated for resistance to ciprofloxacin (CIP), imipenem (IMI), trimetroprim-sulfamethoxazole (TS), ampicillin (AMP) and vancomycin (VAN) by the disc diffusion method. Resistant isolates were further typed with the PhenePlateTM system. Growth assessments were performed with an automated spectrophotometer.Results: The rate of intestinal enterococci resistance was <0.6, 1.3, 1.9 and 13% to VAN, IMI, AMP and CIP respectively. Coliform resistance frequencies were 1.1, 2.2 and 2.2% to CIP, IMI and TS respectively. At two sampling occasions, significantly higher rates of ciprofloxacin resistant enterococci were found and the establishment of a resistant clone in the sewer was indicated by the PhP-analysis. Ciprofloxacin resistant intestinal enterococci had a significantly longer lag-phase time than sensitive isolates, but from 500 µg ml−1 (half MIC) resistant isolates had a competitive advantage in terms of significantly faster generation time.Discussion: Despite high concentration of antimicrobials in the sediment, resistance frequencies were generally low. This can depend on limited growth possibilities for faecal bacteria. However, the establishment of a resistant clone shows that hospital sewers can serve as a reservoir for antibiotic resistant bacteria.