Increased Levels of Multiresistant Bacteria and Resistance Genes after Wastewater Treatment and Their Dissemination into Lake Geneva, Switzerland

Abstract

At present, very little is known about the fate and persistence of multiresistant bacteria (MRB) and their resistance genes in natural aquatic environments. Treated, but partly also untreated sewage of the city of Lausanne, Switzerland is discharged into Vidy Bay (Lake Geneva) resulting in high levels of contamination in this part of the lake. In the present work we have studied the prevalence of MRB and resistance genes in the wastewater stream of Lausanne. Samples from hospital and municipal raw sewage, treated effluent from Lausanne’s wastewater treatment plant (WTP) as well as lake water and sediment samples obtained close to the WTP outlet pipe and a remote site close to a drinking water pump were evaluated for the prevalence of MRB. Selected isolates were identified (16S rRNA gene fragment sequencing) and characterized with regards to further resistances, resistance genes, and plasmids. Mostly, studies investigating this issue have relied on cultivation-based approaches. However, the limitations of these tools are well known, in particular for environmental microbial communities, and cultivation-independent molecular tools should be applied in parallel in order to take non-culturable organisms into account. Here we directly quantified the sulfonamide resistance genes sul1 and sul2 from environmental DNA extracts using TaqMan real-time quantitative PCR. Hospital sewage contained the highest load of MRB and antibiotic resistance genes (ARGs). Wastewater treatment reduced the total bacterial load up to 78% but evidence for selection of extremely multiresistant strains and accumulation of resistance genes was observed. Our data clearly indicated pollution of sediments with ARGs in the vicinity of the WTP outlet. The potential of lakes as reservoirs of MRB and potential risks are discussed.

Wastewater as a point source of antibiotic-resistance genes in the sediment of a freshwater lake

“…Study site description and sampling campaigns For a detailed description of the Vidy Bay study site, please refer to Czekalski et al (2012). Lausanne (population 214 000) is the biggest city in the Lake Geneva area.…”

“…Sediment cores were retrieved as described previously (Czekalski et al, 2012). Sediment cores were transported to the laboratory within 8 h and stored at 4 1C for a maximum of 48 h until processing.…”

“…Micropollutants, including antibiotics (Bonvin et al, 2011), as well as multiresistant bacteria and ARGs pass through the WWTP and are subsequently discharged in considerable amounts into Vidy Bay (Czekalski et al, 2012). Low levels of multiresistant bacteria and ARGs were found even 3.2 km from the WWTP discharge pipe, where lake water is pumped for drinking water production (Czekalski et al, 2012). However, it remained unclear whether this represented contamination or the (unknown) natural resistance background level of Lake Geneva.…”

“…The bay's sediments have been reported to be heavily polluted by fecal-indicator bacteria, heavy metals, nitrogen and phosphorus (Poté et al, 2008). Micropollutants, including antibiotics (Bonvin et al, 2011), as well as multiresistant bacteria and ARGs pass through the WWTP and are subsequently discharged in considerable amounts into Vidy Bay (Czekalski et al, 2012). Low levels of multiresistant bacteria and ARGs were found even 3.2 km from the WWTP discharge pipe, where lake water is pumped for drinking water production (Czekalski et al, 2012).…”

Development and Validation of a Fast Procedure To Analyze Amoxicillin in River Waters by Direct-Injection LC–MS/MS

“…Besides that, they can produce resistance in microbiological lineages, causing serious problems of public health, namely difficulties in treating diseases and imbalance of microbial ecosystems. 3,6,10,11 Recently, the toxic effects of amoxicillin toward algae and aquatic microorganisms were reported. 12,13 Amoxicillin has been detected in several environmental matrices since the mid1990s, when new analytical methods such as liquid chromatography tandem mass spectrometry were developed.…”