Abstract

Fosfomycin is a re-emergent antibiotic known to be effective against severe bacterial infections even when other antibiotics fail. To avoid overuse and thus the risk of new antibiotic resistance, the European Commission has recommended the intravenous use of fosfomycin only when other antibiotic treatments fail. A release of fosfomycin into the environment via wastewater from not only municipalities but also already from the producing pharmaceutical industry can seriously undermine a sustaining therapeutic value. We showed in long-term continuous-mode bioreactor cultivation and by using microbial community flow cytometry, microbial community ecology tools, and cell sorting that the micro-pollutant altered the bacterial wastewater community (WWC) composition within only a few generations. Under these conditions, fosfomycin was not readily degraded both at lower and higher concentrations. At the same time, operational reactor parameters and typical diversity parameters such as α- and intracommunity β-diversity did not point to system changes. Nevertheless, an intrinsic compositional change occurred, caused by a turnover process in which higher concentrations of fosfomycin selected for organisms known to frequently harbor antibiotic resistance genes. A gfp-labeled Pseudomonas putida strain, used as the model organism and a possible future chassis for fosfomycin degradation pathways, was augmented and outcompeted in all tested situations. The results suggest that WWCs, as complex communities, may tolerate fosfomycin for a time, but selection for cell types that may develop resistance is very likely. The approach presented allows very rapid assessment and visualization of the impact of antibiotics on natural or managed microbial communities in general and on individual members of these communities in particular.

Highlights

  • Worldwide, depending on the antibiotic class, 20–80% of antibiotics enter the environment via urine and feces as well as via the deliberate or accidental release (European Medicines Agency, 2020), e.g., by production facilities of the pharmaceutical industry (Andersson and Hughes, 2014)

  • The variations caused by fosfomycin on wastewater community (WWC) structures were monitored using a bioreactor setup operated in a continuous mode

  • Sorted subsets of cells were further investigated by 16S rRNA amplicon sequencing to unravel species that were persistent under the pressure of fosfomycin

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Summary

Introduction

Worldwide, depending on the antibiotic class, 20–80% of antibiotics enter the environment via urine and feces as well as via the deliberate or accidental release (European Medicines Agency, 2020), e.g., by production facilities of the pharmaceutical industry (Andersson and Hughes, 2014). The number of potential antibiotic-resistant pathogens (ARPs) has been found to decrease from the inflow to the effluent in a WWTP, acquired resistance is spreading between wastewater environments and ecosystems (Andersson and Hughes, 2014; Hernando-Amado et al, 2019; Hou et al, 2019). Fosfomycin acts during the early stage of cell wall synthesis both in Gram-positive and Gram-negative bacteria (Aghamali et al, 2019). It is currently considered a valuable re-emergent antibiotic with broad-spectrum antibacterial activity and effective against multidrug-resistant isolates (Cao et al, 2019). In industrial production, similar to the production of other antibiotics, fosfomycin is disposed of in wastewater and inefficiently removed by conventional wastewater treatment systems before being released into the environment. Most of the larger fosfomycin manufacturers rely on biological processes to treat pharmaceutical wastewater

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