Safe surface concept in vertical flow constructed wetland design to mitigate infection hazard.

Decentralized wastewater management based on vertical flow constructed wetlands (VFCWs) can be an effective solution for minimizing sanitation problems also in urban landscapes, especially when considering rapidly expanding cities in developing countries. Yet, the mass implementation of VFCWs in urbanized areas first needs improvement of a few design drawbacks - among them, the control of infection hazard is of primary importance. Therefore, in this study, the possibility of mitigation of the VFCW-derived infection hazard was assessed, through analysis of bacteriostatic properties of top filtration layer materials, according to clinical experiences based on "safe" antimicrobial surfaces. The experiment was carried out on a daily operating VFCW. Coliform bacteria survival rates were measured for known VFCW construction materials such as Pinus bark, gravel, slag, charcoal and LECA. The calculated die-off rates expressed as 12-h first-order inactivation coefficients ranged between 6.91 h-1 (slag/summer) and 0.58 h-1 (Pinus bark/autumn). The obtained die-off curves showed charcoal, Pinus bark and LECA to have little bacteriostatic properties - even occasionally providing conditions promoting the growth of the coliform population. Meanwhile, slag and gravel were strictly inhibiting bacteria growth, reducing the population up to 99% within the first 3 h of contact time. The research showed that it is possible to significantly mitigate the infection hazard of VFCW by means of proper top-layer substrate material, similar or equal to slag or gravel.

Microbial community and antimicrobial resistance niche differentiation in a multistage, surface flow constructed wetland.

Free-living (FL) and particulate-associated (PA) communities are distinct bacterioplankton lifestyles with different mobility and dissemination routes. Understanding spatio-temporal dynamics of PA and FL fractions will allow improvement to wastewater treatment processes including pathogen and AMR bacteria removal. In this study, PA, FL and sediment community composition and antimicrobial resistance gene (ARG; tetW, ermB, sul1, intI1) dynamics were investigated in a full-scale municipal wastewater free-water surface polishing constructed wetland. Taxonomic composition of PA and FL microbial communities shifted towards less diverse communities (Shannon, Chao1) at the CW effluent but retained a distinct fraction-specific composition. Wastewater treatment plant derived PA communities introduced the bulk of AMR load (70 %) into the CW. However, the FL fraction was responsible for exporting over 60 % of the effluent AMR load given its high mobility and the effective immobilization (1-3 log removal) of PA communities. Strong correlations (r2>0.8, p < 0.05) were observed between the FL fraction, tetW and emrB dynamics, and amplicon sequence variants (ASVs) of potentially pathogenic taxa, including Bacteroides, Enterobacteriaceae, Aeromonadaceae, and Lachnospiraceae. This study reveals niche differentiation of microbial communities and associated AMR in CWs and shows that free-living bacteria are a primary escape route of pathogenic and ARG load from CWs under low-flow hydraulic conditions.

Microplastic dynamics in a free water surface constructed wetland.

This study investigates microplastic (MPs) dynamics of a recently established surface flow 2100 population equivalent polishing constructed wetland (CW) receiving 1.4 ML per day of secondary treated wastewater. MPs type, size ranges and concentrations were measured along the CW at a 2-months sampling campaign. The CW received an average of 5·106 MPs per day (6 MPs per liter), mostly 100-1000 µm-sized synthetic fibers followed by fragments in the same size range. 95 % of MPs were retained, resulting in 0.30 ± 0.09 MPs per liter in CW effluent. Most MPs (97 %) were trapped within the first 20 % of the CW which consisted of a settling pond and shallow vegetated treatment cells and provided an areal removal rate > 4000 MP m-2 d-1. Data and microscopic analysis indicate MPs erosion and fragmentation in the CW. Turbidity and suspended solids were no indicator for MP removal due to water fowl activity, algal growth, and preferential flow conditions. This is the first study on MP dynamics in an independently operating full scale free water surface CW incorporated into a municipal wastewater treatment scheme. Surface flow CWs can retain MPs effectively but accumulation in CW sediments and substrate needs to be considered when further utilized or recycled.

Microplastic biofilm, associated pathogen and antimicrobial resistance dynamics through a wastewater treatment process incorporating a constructed wetland.

Microplastics in wastewater are colonized by biofilms containing pathogens and antimicrobial resistance (AMR) genes that can be exported into receiving water bodies. This study investigated establishment and changes in microplastic-associated biofilm and AMR during a conventional full-scale 2100 population equivalent wastewater treatment process combined with a free water surface polishing constructed wetland. Sequential microplastic colonization experiments were conducted at different stages of the wastewater treatment process, including in raw sewage, treated effluent and the constructed wetland. Two scenarios were tested in which the constructed wetland served as either (i) a polishing step or (ii) as primary recipient of sewage inoculated microplastics. Bacterial 16S rRNA gene sequencing was carried out for qualitative bacterial community analysis. qPCR was applied for quantitative analysis of AMR genes (sul1, ermB, tetW, intiI1), bacterial biomass (16S rRNA) and a human fecal marker (HF183). Microbial diversity on microplastics increased with incubation time. The initial sewage-derived biofilm composition changed more significantly in the wastewater effluent compared to the constructed wetland. Pathogen and AMR load decreased by up to two orders of magnitude after coupled conventional and constructed wetland treatment, while less impact was observed when sewage-inoculated microplastic material was directly transferred into the constructed wetland. Aeromonas, Klebsiella, and Streptococcus were key pathogenic genera correlated with AMR in microplastic-associated biofilms. Despite decreasing trends on human pathogens and AMR load along the treatment process, microplastic-associated biofilms were a considerable potential hotspot for AMR (intI1 gene) and accommodated Cyanobacteria and fish pathogens.