Assessing antibiotic resistance of microorganisms in sanitary sewage

The release of antimicrobial substances into surface waters is of growing concern due to direct toxic effects on all trophic levels and the promotion of antibiotic resistance through sub-inhibitory concentration levels. This study showcases (1) the variation of antibiotics in sanitary sewage depending on different timescales and (2) a method to assess the antibiotic resistance based on an inhibition test. The test is based on the measurement of the oxygen uptake rate (OUR) in wastewater samples with increasing concentrations of the selected antibiotic agents. The following antibiotics were analysed in the present study: clarithromycin (CLA) was selected due to its high toxicity to many microorganisms (low EC50), ciprofloxacin (CIP) which is used to generally fight all bacteria concerning interstitial infections and doxycyclin (DOX) having a broad spectrum efficacy. Results show that CLA inhibited the OUR by approximately 50% at a concentration of about 10 mg L−1, because Gram-negative bacteria such as Escherichia coli are resistant, whereas CIP inhibited about 90% of the OUR at a concentration equal to or greater than 10 mg L−1. In the case of DOX, a moderate inhibition of about 38% at a concentration of 10 mg L−1 was identified, indicating a significant antibiotic resistance. The results are consistent with the corresponding findings from the Clinical and Laboratory Standards Institute. Thus, the presented inhibition test provides a simple but robust alternative method to assess antibiotic resistance in biofilms instead of more complex clinical tests.