Identification of Antibiotic Resistance Gene Hosts in Treatment Wetlands Using a Single-Cell Based High-Throughput Approach

Determining the prevalence of antimicrobial resistance (AMR) in non-clinical settings is vital for better management of the global AMR crisis. Untreated and even treated wastewaters are important sources that release AMR into the environment. Methodologically, it is difficult to generate a comprehensive in situ profile of antibiotic resistance gene hosts. Here, we used epicPCR (emulsion, paired isolation, and concatenation PCR) as a cultivation-independent method to reveal the host profiles of the AMR indicator genes intI1, sul1, sul2, and dfrA1 in two constructed wetlands treating municipal wastewater. Overall, the epicPCR analysis revealed a profile of AMR indicator gene hosts that is consistent with literature data from cultivation-based approaches. Most carriers of antibiotic resistance (AR) genes and likely of class 1 integrons belonged to the Gammaproteobateria, particularly the Burkholderiaceae and Rhodocyclaceae families, followed by members of the Campylobacterota, Desulfobacterota, and Firmicutes. The analysis also identified several novel hosts for the indicator genes widely distributed in the wetlands, including the genera Legionella and Ralstonia. Therefore, the application of epicPCR has produced an expanded insight into the in situ indicator gene host profile, while highlighting the role of the environment as a reservoir for AMR

Treatment wetlands

Treatment Wetlands is the seventh volume in the Biological Wastewater Treatment series, which gives a state-of-the-art presentation of the science and technology of sewage treatment. The major variants of wetland systems are covered in this volume, namely: (i) horizontal flow wetlands; (ii) vertical flow wetlands; (iii) French vertical flow wetlands; (iv) intensified wetlands; (v) free water surface wetlands; (vi) other applications of treatment wetlands. The book presents in a clear and didactic way the main concepts, working principles, expected performance, design criteria, design examples, construction aspects and operational guidelines. The book has been written by an international team of top experts in the field of treatment wetlands.Postprint (published version

Treatment Wetlands

Overview of Treatment Wetlands; Fundamentals of Treatment Wetlands; Horizontal Flow Wetlands; Vertical Flow Wetlands; French Vertical Flow Wetlands; Intensified and Modified Wetlands; Free Water Surface Wetlands; Other Applications; Additional Aspects

Resilience of Micropollutant and Biological Effect Removal in an Aerated Horizontal Flow Treatment Wetland

The performance of an aerated horizontal subsurface flow treatment wetland was investigated before, during and after a simulated aeration failure. Conventional wastewater parameters (e.g., carbonaceous biological oxygen demand, total nitrogen, and Escherichia coli) as well as selected micropollutants (caffeine, ibuprofen, naproxen, benzotriazole, diclofenac, acesulfame, and carbamazepine) were investigated. Furthermore, the removal of biological effects was investigated using in vitro bioassays. The six bioassays selected covered environmentally relevant endpoints (indicative of activation of aryl hydrocarbon receptor, AhR; binding to the peroxisome proliferator-activated receptor gamma, PPARγ; activation of estrogen receptor alpha, ERα; activation of glucocorticoid receptor, GR; oxidative stress response, AREc32; combined algae test, CAT). During the aeration interruption phase, the water quality deteriorated to a degree comparable to that of a conventional (non-aerated) horizontal subsurface flow wetland. After the end of the aeration interruption, the analytical and biological parameters investigated recovered at different time periods until their initial treatment performance. Treatment efficacy for conventional parameters was recovered within a few days, but no complete recovery of treatment efficacy could be observed for bioassays AhR, AREc32 and CAT in the 21 days following re-start of the aeration system. Furthermore, the removal efficacy along the flow path for most of the chemicals and bioassays recovered as it was observed in the baseline phase. Only for the activation of AhR and AREc32 there was a shift of the internal treatment profile from 12.5% to 25% (AhR) and 50% (AREc32) of the fractional length

Recent Advances in the Application, Design, and Operations & Maintenance of Aerated Treatment Wetlands

This paper outlines recent advances in the design, application, and operations and maintenance (O&M) of aerated treatment wetland systems as well as current research trends. We provide the first-ever comprehensive estimate of the number and geographical distribution of aerated treatment wetlands worldwide and review new developments in aerated wetland design and application. This paper also presents and discusses first-hand experiences and challenges with the O&M of full-scale aerated treatment wetland systems, which is an important aspect that is currently not well reported in the literature. Knowledge gaps and suggestions for future research on aerated treatment wetlands are provided

Reversing clogging in subsurface-flow constructed wetlands by hydrogen peroxide treatment: two case studies

One of the most frequently encountered operational problems in subsurface-flow constructed wetlands is clogging. Traditionally, the restoration procedure is to remove the clogged gravel or sand and replace it with clean material. This method, while effective, is costly and may require sections of the facility to be taken offline for extended periods of time. Another common remediation strategy is to have a resting period for each wetland cell, although this is not an option for very small systems which often consist of only one treatment cell. Recently, a more radical approach has been tested on a number of lab-scale and pilot-scale setups which consists of an aggressive oxidation of organic matter by means of hydrogen peroxide (H(2)O(2)). Results indicate that after treatment, clogging was substantially reduced and that H(2)O(2) did not appear to have a long-term negative effect on plants and biofilms. The outcomes of two full-scale tests are discussed in this paper

Recent Advances in the Application, Design, and Operations & Maintenance of Aerated Treatment Wetlands

International audienceThis paper outlines recent advances in the design, application, and operations and maintenance (O&M) of aerated treatment wetland systems as well as current research trends. We provide the first-ever comprehensive estimate of the number and geographical distribution of aerated treatment wetlands worldwide and review new developments in aerated wetland design and application. This paper also presents and discusses first-hand experiences and challenges with the O&M of full-scale aerated treatment wetland systems, which is an important aspect that is currently not well reported in the literature. Knowledge gaps and suggestions for future research on aerated treatment wetlands are provided

What is the best procedure for determining removal rate coefficients in horizontal flow treatment wetlands: influent and effluent concentrations or longitudinal concentration profiles?

First-order removal rate coefficients (k) are used in predictive equations for estimating effluent concentrations from horizontal flow (HF) wetlands. Due to limited resources, influent and effluent concentration data from existing systems are frequently used in the estimation of k values from operating systems, but another choice is to use concentration data along the longitudinal profile of the HF wetland. Based on a dataset with 41 HF wetlands/studies obtained from a literature survey, with chemical oxygen demand (COD) measurements at different sampling points, volumetric (kV) and areal (kA) removal rate coefficients for the Tanks-In-Series (TIS) model have been obtained using the two estimation methods. In general, removal rate coefficients derived from longitudinal profiles of concentrations were higher than those obtained by using data from influent and effluent concentrations, reflecting the fact that constituent removal is mostly accomplished before the wastewater reaches the outlet zone. Deriving coefficients from longitudinal profiles is more comprehensive, providing a better explanation of the internal removal taking place in the treatment wetland. However, the more widely used approach of calculating kV and kA from influent/effluent concentrations may lead to a safer design of horizontal flow wetlands, because of underestimation of the actual removal rate coefficients. HIGHLIGHTS Lack of information on best way to derive first-order removal rate coefficients.; Influent/effluent concentrations or longitudinal profile of concentrations are used.; Database with 41 HF wetlands was constructed and used for calculation of k values.; Volumetric and areal k values from longitudinal profiles method were usually higher.; k values from influent/effluent data will result in a more conservative design.

Do wastewater pollutants impact oxygen transfer in aerated horizontal flow wetlands?

International audienceAerated treatment wetlands are an increasingly recognized nature-based technology for wastewater treatmentthat relies heavily on mechanical aeration. Although aeration-mediated oxygen transfer into the wastewater canbe impeded by wastewater pollutants, little is known about the link between the volumetric oxygen masstransfer coefficient and the organic carbon concentration of the wastewater in aerated wetlands. In this study,oxygen transfer experiments were carried out in a lab-scale gravel column using clean water and wastewaterfrom a pilot-scale horizontal flow (HF) aerated wetland treating domestic sewage. The-factor, which describesthe ratio of the volumetric oxygen mass transfer coefficient in wastewater to clean water, was reduced byincreasing soluble chemical oxygen demand (CODs). The derived regression equation21.0661.372E-3 mg COD Ls1was incorporated into a numerical process model to simulate the impact of thereduced oxygen transfer on a hypothetical HF aerated wetland. The simulations revealed thatand treatmentefficacy for nitrogen were substantially reduced by CODsat low aeration ( of 1 h−1) and high influent waste-water strength (CODsof 300 mg L−1). At the same and influent CODsconcentration, longitudinal gradients ofand concentrations for dissolved oxygen (DO), NH4-N and NOx-N in the simulated wetland were shifted up to21% of wetland length downstream. These effects decreased with increasing and were found to be negligible at> 3 h−1, which corresponds to an air flow rate of approximately 400 L m−2h−1. Following this, higher organiccarbon concentrations can reduce oxygen transfer in HF aerated wetland systems, thus resulting in decreasedtreatment efficacy