The Impact of the Antibiotic Fosfomycin on Wastewater Communities Measured by Flow Cytometry

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.

Development of an Automated Online Flow Cytometry Method to Quantify Cell Density and Fingerprint Bacterial Communities

“…Flow cytometry has already been used in the past to monitor bacterial communities to detect dynamic changes in their composition [ 27 , 46 , 47 ]. HNA/LNA ratios were used widely in flow cytometry in the past for the determination of two subsets of communities, taking up either low or high amounts of SYBR Green [ 34 , 48 ].…”

Nanosecond pulsed discharge with plasma-functionalized TiO2 for the high efficiency degradation of tetracycline

“…The toxicity of the NPG-LD/TiO 2 -P treated solution is almost the same as that of deionized water, showing that the toxicity of the treated solution was reduced after plasma degradation, and the NPG-LD/TiO 2 -P system could further reduce the toxicity of antibiotics. In order to quantitatively analyze the state of cell apoptosis, figure 10(b) presents the variation of HCT116 cells in the treated solution detected by the flow cytometry [45]. The apoptosis rate of HCT116 cells in the untreated tetracycline solution reached 43.5%, and those in the NPG-LD, NPG-LD/TiO 2 and NPG-LD/TiO 2 -P systems were 37.45%, 9.02% and 8.48%, respectively.…”

Current and Future Flow Cytometry Applications Contributing to Antimicrobial Resistance Control

“…Data integration from FCM analysis, protein analysis, and DNA sequencing could contribute to a comprehensive understanding of microbial communities' responses to antibiotics. Li et al (2022) [103] reported the usefulness of high throughput FCM and 16S rRNA gene sequencing in assessing the impact of low fosfomycin pressure over long periods of time on microbial communities from wastewaters, such information is of importance to both control spread and resistance evolution as these wastewaters are used on farmland [104]. Successful application of FCM cell counting in the evaluation of wastewater treatment processes has been reported [58,105,106].…”