Assessment of sulfamethoxazole and oxytetracycline uptake and transformation in Eisenia fetida earthworms

The scientific community is becoming increasingly concerned about the recent detection of transformation products (TPs) of antimicrobials (AMs) and their presence in the food chain. There are growing concerns about the potential consequences on food safety and the proliferation of antimicrobial resistance. In this work, the transformation process of sulfamethoxazole (SMX) and oxytetracycline (OTC) in soil was thoroughly evaluated. For that purpose, soils were homogeneously contaminated at three concentration levels of SMX and OTC, independently, and samples were analysed after 7 and 14 days by Ultra High-Performance Liquid Chromatography coupled to a triple quadrupole mass spectrometer (UHPLC-MS/MS). The results have demonstrated a remarkable transformation, particularly noteworthy for SMX, as it exhibited an 89 % - 94 % decrease in concentration within the initial 7 days of the experiment. In addition, to assess whether terrestrial organisms would be able to accumulate the AMs, Eisenia fetida (E. fetida) earthworms were exposed to the above-mentioned concentration levels of AMs in soil. Both AMs were accumulated in the organisms after 14 days, but higher bioaccumulation factor values (BCF) were determined for SMX (0.52-17.84) compared to OTC (0.02-0.21) at all tested concentrations. The analyses were extended to search for TPs in earthworms and soils using a suspect screening approach. Concretely, by means of UHPLC-high resolution mass spectrometry (UHPLC-HRMS) three TPs were identified at 2a and 2b of confidence level. To the best of our knowledge, one SMX-TP and one OTC-TP were identified in earthworms and soil, respectively, for the first time in the present work. Earthworms did not experience weight loss or mortality in the presence of these AMs at levels found in the environment, but there was a decrease in riboflavin levels, which is linked to changes in the immune system. This study represents a significant advancement in understanding the impact of AMs in soil and their subsequent entry into the food chain. It also provides valuable insights into the potential effects of AMs and their TPs on organisms.