MiDAS 5: Global diversity of bacteria and archaea in anaerobic digesters.

Anaerobic digestion of organic waste into methane and carbon dioxide (biogas) is carried out by complex microbial communities. Here, we use full-length 16S rRNA gene sequencing of 285 full-scale anaerobic digesters (ADs) to expand our knowledge about diversity and function of the bacteria and archaea in ADs worldwide. The sequences are processed into full-length 16S rRNA amplicon sequence variants (FL-ASVs) and are used to expand the MiDAS 4 database for bacteria and archaea in wastewater treatment systems, creating MiDAS 5. The expansion of the MiDAS database increases the coverage for bacteria and archaea in ADs worldwide, leading to improved genus- and species-level classification. Using MiDAS 5, we carry out an amplicon-based, global-scale microbial community profiling of the sampled ADs using three common sets of primers targeting different regions of the 16S rRNA gene in bacteria and/or archaea. We reveal how environmental conditions and biogeography shape the AD microbiota. We also identify core and conditionally rare or abundant taxa, encompassing 692 genera and 1013 species. These represent 84-99% and 18-61% of the accumulated read abundance, respectively, across samples depending on the amplicon primers used. Finally, we examine the global diversity of functional groups with known importance for the anaerobic digestion process.

Sewershed surveillance as a tool for smart management of a pandemic in threshold countries. Case study: Tracking SARS-CoV-2 during COVID-19 pandemic in a major urban metropolis in northwestern Argentina.

Wastewater-based epidemiology is an economical and effective tool for monitoring the COVID-19 pandemic. In this study we proposed sampling campaigns that addressed spatial-temporal trends within a metropolitan area. This is a local study of detection and quantification of SARS-CoV-2 in wastewater during the onset, rise, and decline of COVID-19 cases in Salta city (Argentina) over the course of a twenty-one-week period (13 Aug to 30 Dec) in 2020. Wastewater samples were gathered from 13 sewer manholes specific to each sewershed catchment, prior to convergence or mixing with other sewer lines, resulting in samples specific to individual catchments with defined areas. The 13 sewershed catchments selected comprise 118,832 connections to the network throughout the city, representing 84.7 % (534,747 individuals) of the total population. The number of COVID19-related exposure and symptoms cases in each area were registered using an application developed for smartphones by the provincial government. Geographical coordinates provided by the devices were recorded, and consequently, it was possible to geolocalise all app-cases and track them down to which of the 13 sampling catchments belonged. RNA fragments of SARS-CoV-2 were detected in every site since the beginning of the monitoring, anticipating viral circulation in the population. Over the course of the 21-week study, the concentrations of SARS-CoV-2 ranged between 1.77 × 104 and 4.35 × 107 genome copies/L. There was a correspondence with the highest viral load in wastewater and the peak number of cases reported by the app for each catchment. The associations were evaluated with correlation analysis. The viral loads of SARS-CoV-2 in wastewater were a feasible means to describe the trends of COVID-19 infections. Surveillance at sewershed scale, provided reliable and strategic information that could be used by local health stakeholders to manage the COVID-19 pandemic.

Selective Enrichment of Nitrososphaera viennensis-Like Ammonia-Oxidizing Archaea over Ammonia-Oxidizing Bacteria from Drinking Water Biofilms.

Ammonia-oxidizing archaea (AOA) can oxidize ammonia to nitrite for energy gain. They have been detected in chloraminated drinking water distribution systems (DWDS) along with the more common ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). To date, no members of the AOA have been isolated or enriched from drinking water environments. To begin the investigation of the role of AOA in chloraminated DWDS, we developed a selective approach using biofilm samples from a full-scale operational network as inoculum. A Nitrososphaera viennensis-like AOA taxon was enriched from a mixed community that also included Nitrosomonas-like AOB while gradually scaling up the culture volume. Dimethylthiourea (DMTU) and pyruvate at 100 µM were added to promote the growth of AOA while inhibiting AOB. This resulted in the eventual washout of AOB, while NOB were absent after 2 or 3 rounds of amendment with 24 µM sodium azide. The relative abundance of AOA in the enrichment increased from 0.2% to 39.5% after adding DMTU and pyruvate, and further to 51.6% after filtration through a 0.45-µm pore size membrane, within a period of approximately 6 months. IMPORTANCE Chloramination has been known to increase the risk of nitrification episodes in DWDS due to the presence of ammonia-oxidizing microorganisms. Among them, AOB are more frequently detected than AOA. All publicly available cultures of AOA have been isolated from soil, marine or surface water environments, meaning they are allochthonous to DWDS. Hence, monochloramine exposure studies involving these strains may not accurately reflect their role in DWDS. The described method allows for the rapid enrichment of autochthonous AOA from drinking water nitrifying communities. The high relative abundance of AOA in the resulting enrichment culture reduces any confounding effects of co-existing heterotrophic bacteria when investigating the response of AOA to varied levels of monochloramine in drinking water.

Tracking SARS-CoV-2 in rivers as a tool for epidemiological surveillance.

The aim of this work was to evaluate if rivers could be used for SARS-CoV-2 surveillance. Five sampling points from three rivers (AR-1 and AR-2 in Arenales River, MR-1 and MR-2 in Mojotoro River, and CR in La Caldera River) from Salta (Argentina), two of them receiving discharges from wastewater plants (WWTP), were monitored from July to December 2020. Fifteen water samples from each point (75 in total) were collected and characterized physico-chemically and microbiologically and SARS-CoV-2 was quantified by RT-qPCR. Also, two targets linked to human contributions, human polyomavirus (HPyV) and RNase P, were quantified and used to normalize SARS-CoV-2 concentration, which was compared to reported COVID-19 cases. Statistical analyses allowed us to verify the correlation between SARS-CoV-2 and the concentration of fecal indicator bacteria (FIB), as well as to find similarities and differences between sampling points. La Caldera River showed the best water quality; FIBs were within acceptable limits for recreational activities. Mojotoro River's water quality was not affected by the northern WWTP of the city. Instead, Arenales River presented the poorest water quality; at AR-2 was negatively affected by the discharges of the southern WWTP, which contributed to significant increase of fecal contamination. SARS-CoV-2 was found in about half of samples in low concentrations in La Caldera and Mojotoro Rivers, while it was high and persistent in Arenales River. No human tracers were detected in CR, only HPyV was found in MR-1, MR-2 and AR-1, and both were quantified in AR-2. The experimental and normalized viral concentrations strongly correlated with reported COVID-19 cases; thus, Arenales River at AR-2 reflected the epidemiological situation of the city. This is the first study showing the dynamic of SARS-CoV-2 concentration in an urban river highly impacted by wastewater and proved that can be used for SARS-CoV-2 surveillance to support health authorities.

Nitrifying niche differentiation in biofilms from full-scale chloraminated drinking water distribution system.

Tropical conditions favour the auto-decomposition of monochloramine (MCA) leading to disinfectant decay and free ammonia in drinking water distribution systems (DWDS); thus, they promote the growth of nitrifiers and the development of biofilms on the inner-pipe surface. Biofilms can adversely impact the provision of safe and biologically stable water. Moreover, there is a general lack of understanding of the role of microbial communities in DWDS in regions with warm temperatures and no distinct seasons. Here, we report a survey on biofilms from full-scale monochloraminated DWDS in a highly urbanised metropolis using next generation sequencing tools. The monitoring campaign consisted of sampling biofilms and bulk waters from 21 in-service pipes. We characterized the microbial community with emphasis on nitrifying bacteria and archaea using 16S rRNA gene amplicon sequencing and potential nitrification activity. Samples grouped into two clusters, characterized by their low (Cluster LD) and high (Cluster HD) α-diversity. Both clusters harbour microorganisms related to nitrification: i) Nitrosomonas (24.9-68.8%), an ammonia oxidising bacterium (AOB) that dominated Cluster LD, and ii) a co-aggregation of genus Nitrospira (9.8-32.5%), a nitrite oxidising bacterium (NOB), and Thaumarchaeota (1.4-10.9%), chemolithotrophic ammonia oxidising (AOA) archaea that were among the most abundant OTUs in Cluster HD. Activity tests performed with fresh biofilm samples confirmed that these two clusters represent distinctive biofilm niches performing different stages of the nitrification process. Cluster LD correlated with a high concentration of MCA, which caused dysbiosis and resulted in high unevenness of the cluster. In cluster HD, with more biomass, chemical reactions involving nitrite increased the MCA demand, releasing ammonia and allowing more nitrifiers to grow, like AOA and NOB. From this study, we conclude that an MCA residual gradient along the DWDS drives and shapes the microbial community assembly and should be considered when designing effective disinfection strategies.

Agricultural contamination impacts antibiotic resistance gene abundances in river bed sediment temporally.

Kewaunee County, Wisconsin is an agricultural area dominated by concentrated animal feeding operations and manure fertilized cropland. The objective of this study was to characterize chemical and antibiotic resistance gene (ARG) profiles of 20 surface water locations in Kewaunee County to better understand relationships between agricultural contamination and ARG abundance over one year. Surface water (n = 101) and bed sediment (n = 93) were collected from 20 sites during five timepoints between July 2016 and May 2017. Samples were analyzed for six genes (erm(B), tet(W), sul1, qnrA, intI1 and 16S rRNA) and water chemistry and pollution indicators. qnrA, intI1 and sul1 genes in surface water were significantly higher than erm(B) and tet(W); however, no difference was present in sediment samples. Redundancy analysis identified positive correlations of nitrate, Escherichia coli, and coliforms with tet(W) and intI1 genes in sediment and intI1, sul1 and tet(W) genes in water. Temporal patterns of ARG abundance were identified with significantly higher gene abundances found in sediment during Kewaunee County's manure fertilization period; however, surface water patterns were not distinct. Together, these results suggest Kewaunee County sediments serve as a site of accumulation for non-point source agricultural pollution and ARGs on a temporal scale associated with manure fertilization.

Plasma deposition of silver nanoparticles on ultrafiltration membranes: antibacterial and anti-biofouling properties.

A novel and versatile plasma reactor was used to modify Polyethersulphone commercial membranes. The equipment was applied to: i) functionalize the membranes with low-temperature plasmas, ii) deposit a film of poly(methyl methacrylate) (PMMA) by Plasma Enhanced Chemical Vapor Deposition (PECVD) and, iii) deposit silver nanoparticles (SNP) by Gas Flow Sputtering. Each modification process was performed in the same reactor consecutively, without exposure of the membranes to atmospheric air. Scanning electron microscopy and transmission electron microscopy were used to characterize the particles and modified membranes. SNP are evenly distributed on the membrane surface. Particle fixation and transport inside membranes were assessed before- and after-washing assays by X-ray photoelectron spectroscopy depth profiling analysis. PMMA addition improved SNP fixation. Plasma-treated membranes showed higher hydrophilicity. Anti-biofouling activity was successfully achieved against Gram-positive (Enterococcus faecalis) and -negative (Salmonella Typhimurium) bacteria. Therefore, disinfection by ultrafiltration showed substantial resistance to biofouling. The post-synthesis functionalization process developed provides a more efficient fabrication route for anti-biofouling and anti-bacterial membranes used in the water treatment field. To the best of our knowledge, this is the first report of a gas phase condensation process combined with a PECVD procedure in order to deposit SNP on commercial membranes to inhibit biofouling formation.

Strategies to optimize monitoring schemes of recreational waters from Salta, Argentina: a multivariate approach.

Several recreational surface waters in Salta, Argentina, were selected to assess their quality. Seventy percent of the measurements exceeded at least one of the limits established by international legislation becoming unsuitable for their use. To interpret results of complex data, multivariate techniques were applied. Arenales River, due to the variability observed in the data, was divided in two: upstream and downstream representing low and high pollution sites, respectively, and cluster analysis supported that differentiation. Arenales River downstream and Campo Alegre Reservoir were the most different environments, and Vaqueros and La Caldera rivers were the most similar. Canonical correlation analysis allowed exploration of correlations between physicochemical and microbiological variables except in both parts of Arenales River, and principal component analysis allowed finding relationships among the nine measured variables in all aquatic environments. Variable's loadings showed that Arenales River downstream was impacted by industrial and domestic activities, Arenales River upstream was affected by agricultural activities, Campo Alegre Reservoir was disturbed by anthropogenic and ecological effects, and La Caldera and Vaqueros rivers were influenced by recreational activities. Discriminant analysis allowed identification of subgroup of variables responsible for seasonal and spatial variations. Enterococcus, dissolved oxygen, conductivity, E. coli, pH, and fecal coliforms are sufficient to spatially describe the quality of the aquatic environments. Regarding seasonal variations, dissolved oxygen, conductivity, fecal coliforms, and pH can be used to describe water quality during dry season, while dissolved oxygen, conductivity, total coliforms, E. coli, and Enterococcus during wet season. Thus, the use of multivariate techniques allowed optimizing monitoring tasks and minimizing costs involved.

The impact of point source pollution on shallow groundwater used for human consumption in a threshold country.

Many developing and threshold countries rely on shallow groundwater wells for their water supply whilst pit latrines are used for sanitation. We employed a unified strategy involving satellite images and environmental monitoring of 16 physico-chemical and microbiological water quality parameters to identify significant land uses that can lead to unacceptable deterioration of source water, in a region with a subtropical climate and seasonally restricted torrential rainfall in Northern Argentina. Agricultural and non-agricultural sources of nitrate were illustrated in satellite images and used to assess the organic load discharged. The estimated human organic load per year was 28.5 BOD(5) tons and the N load was 7.5 tons, while for poultry farms it was 9940-BOD(5) tons and 1037-N tons, respectively. Concentrations of nitrates and organics were significantly different between seasons in well water (p values of 0.026 and 0.039, respectively). The onset of the wet season had an extraordinarily negative impact on well water due in part to the high permeability of soils made up of fine gravels and coarse sand. Discriminant analysis showed that land uses had a pronounced seasonal influence on nitrates and introduced additional microbial contamination, causing nitrification and denitrification in shallow groundwater. P-well was highly impacted by a poultry farm while S-well was affected by anthropogenic pollution and background load, as revealed by Principal Component Analysis. The application of microbial source tracking techniques is recommended to corroborate local sources of human versus animal origin.