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Greywater often contains microorganisms carrying antimicrobial resistance genes (ARGs). Reuse of greywater thus potentially facilitates the enrichment and spread of multidrug resistance, posing a possible hazard for communities that use it. As water reuse becomes increasingly necessary, it is imperative to determine how greywater treatment impacts ARGs. In this study, we characterize ARG patterns in greywater microbial communities before and after treatment by a recirculating vertical flow constructed wetland (RVFCW). This greywater recycling method has been adopted by some small communities and households for greywater treatment; however, its ability to remove ARGs is unknown. We examined the taxonomic and ARG compositions of microbial communities in raw and treated greywater from five households using shotgun metagenomic sequencing. Total ARGs decreased in abundance and diversity in greywater treated by the RVFCW. In parallel, the microbial communities decreased in similarity in treated greywater. Potentially pathogenic bacteria associated with antimicrobial resistance and mobile genetic elements were detected in both raw and treated water, with a decreasing trend after treatment. This study indicates that RVFCW systems have the potential to mitigate antimicrobial resistance-related hazards when reusing treated greywater, but further measures need to be taken regarding persistent mobile ARGs and potential pathogens.

Two oil spills occurred in the Evrona Nature Reserve (southern Israel), in 1975 and 2014. This oil contamination induced highly persistent soil hydrophobicity. The objective of this study was to investigate the decrease in oil-induced soil hydrophobicity under different environmental conditions and to assess the relationship between the hydrophobicity and hydrocarbon content. A laboratory incubation experiment was conducted over 1.5 years to monitor the soil hydrophobicity and total hydrocarbon concentration under different environmental conditions. We hypothesized that the addition of water (20% or 50% saturation), nutrients, and biosurfactants can accelerate the reduction in hydrophobicity and decomposition of hydrocarbons. Water drop penetration time and molarity of ethanol droplet tests were used to assess soil hydrophobicity. In parallel, alkane composition and total petroleum hydrocarbons were evaluated to indicate oil attenuation. The addition of water, nutrients, and biosurfactants resulted in a concomitant reduction in hydrophobicity and hydrocarbon concentration of varying degrees, exhibiting enhanced degradation and hydrophobicity reduction observed in treatments to which nutrients and biosurfactants were added. At the end of the incubation, however, soil hydrophobicity in all treatments remained severe, even though total petroleum hydrocarbon removal was fairly high and reached 40%–80% in the treatments to which water with or without nutrients and/or surfactants was added.

In-situ bioremediation techniques are cost-effective, environmentally friendly, and sustainable. This study examined a large-scale in-situ treatment of an unsaturated zone and a local groundwater system that were heavily contaminated with perchlorate and other co-contaminants, including nitrate, chlorate, and RDX. Principally, the upper section of the unsaturated zone was used as a bioreactor for treating the deep unsaturated zone and groundwater. The treatment was based on a cyclic process that included pumping contaminated groundwater, adding an essential electron donor, and injecting the amended water back into the top-soil, which was used as a bioreactor in the treatment process. In the shallow soil, the local bacteria reduced the perchlorate to chloride and water, and the treated water continued to displace the major pollutants from the deep part of the vadose zone, where the biological potential for contaminant degradation is low, towards the water table. The contaminated leachates were pumped back to the surface with polluted groundwater as part of the cyclic treatment process. Results show that the other co-contaminants, including nitrate, chlorate, and RDX, were removed. Water flow and reactive transport models were calibrated and validated against a time series of the water contents and bromide and perchlorate concentrations that were obtained across the unsaturated zone using the VMS. The calibrated models enabled quantifying the clean-up process and estimating the required time for full perchlorate removal. According to the model's predictions, after 700 days of continuous operation, all the perchlorate, in a total amount of 7754 kg, would be removed from the unsaturated zone. To obtain full removal, the modelling simulations suggest that the in-situ bioremediation should be implemented for an additional 200 days. Ultimately, we present a low-cost, efficient method for treating perchlorate contamination and potentially that of other pollutants in the subsurface.

Microbial communities in cultivated soils control the fate of pollutants associated with agricultural practice. The present study was designed to explore the response of bacterial communities to the application of the widely-used herbicide atrazine in three different crop fields that differ significantly in their physicochemical structure and nutritional content: the nutrient-rich (with relatively high carbon and nitrogen content) Newe Yaar (NY) and Ha-Ogen (HO) soils and the nutrient-poor, sandy Sde-Eliyahu (SE) soil. The 16 S rRNA gene amplicon sequencing revealed the nutrient poor HO soil differs in its response to atrazine in comparison to the two nutrient-rich soils both in the shortest persistence of atrazine and its effect on community structure and composition. Potential reported bacterial degraders of atrazine such as Pseudomonas, Clostridium and Bacillus were more abundant in contaminated sandy/poor soils (HO) whereas bacteria known for nitrogen cycling such as Azospirillum, Sinorhizobium, Nitrospira and Azohydromonas were significantly more abundant in the nutrient rich contaminated SE soils. No significant increase of potential indigenous degrader Arthrobacter was detected in SE and NY soils whereas a significant increase was recorded with HO soils. An overall shift in bacterial community composition following atrazine application was observed only in the nutrient poor soil. Understanding atrazine persistence and microbiome response to its application of in dependence with soil types serve the design of precision application strategies.

TNT, or 2,4,6-trinitrotoluene, is a common explosive that can contaminate soil and groundwater in production sites, military training areas, and disposal locations. The compound is highly toxic; therefore, there is an urgent need to rehabilitate the impacted environments. Harnessing the microbial ability to biodegrade TNT into environmentally harmless compound(s) is one approach to remediating contaminated sites. In our study, we report on the genomic and metabolic ability of Stenotrophomonas strain SG1 to degrade TNT under aerobic and anaerobic conditions. The bacterial strain SG1 was first isolated as a contaminant from a culture of Diaphorobacter sp. strain DS2 over minimal media supplemented with TNT. The draft genome assembly of strain SG1 is ∼4.7 Mb and is distributed among 358 contigs. The homology search against the custom database of enzymes responsible for TNT biodegradation revealed the presence of three N-ethylmaleimide reductases (NemA) with a defined KEGG ortholog and KEGG pathway of TNT degradation. The presence of respiratory nitrate reductases has also been mapped, which supports denitrification under anaerobic conditions. Experimentally, the TNT transformation rate accelerated when carbon sources, such as sodium acetate, sodium citrate, sodium succinate, sucrose, and glucose (final concentration of 5 mM), were added. Citrate promoted the highest growth and TNT transformation ratio (88.35%) in 120 h. With the addition of 5 mM ammonium chloride, TNT completely disappeared in the citrate and sucrose-containing treatments in 120 h. However, higher biomass was obtained in the sucrose and glucose-containing treatments in 120 h. During incubation, the formation of amino dinitrotoluene isomers, dinitrotoluene isomers, trinitrobenzene, azoxy isomers, diaryl hydroxylamines, and corresponding secondary amines was confirmed by GC/MS and UPLC/MS. 2-Amino-4-nitrotoluene, 4-amino-2-nitrotoluene, and 2-amino-6-nitrotoluene were also identified in the culture supernatant by GC/MS. Under anaerobic conditions, TNT completely disappeared in the citrate and citrate plus nitrate treatments. Since the strain shows the ability to remove TNT, this research should be useful in basic research and practical applications for removing TNT from wastewater.

Municipal sewage carries degraded and intact viral particles and RNA (ribonucleic acid) of SARS-CoV-2 (severe acute respiratory coronavirus 2), shed by COVID-19 (coronavirus disease 2019) patients, to sewage and eventually to wastewater treatment plants. Proper wastewater treatment can prevent uncontrolled discharges of the virus into the environment. However, the role of different wastewater treatment stages in reducing viral RNA concentrations is, thus far, unknown. Here, we quantified SARS-CoV-2 RNA in raw sewage and during the main stages of the activated sludge process from two wastewater treatment plants in Israel, on three different days during the 2020 COVID-19 outbreak. To reduce the detection limit, samples were concentrated prior to quantification by real-time polymerase chain reaction by a factor of 2–43 using ultrafiltration. On average, ∼1 log RNA removal was attained by each of the primary and secondary treatment steps; however, >100 copies of SARS-CoV-2 RNA/mL remained in the secondary effluents. Following chlorination, SARS-CoV-2 RNA was detected only once, likely due to an insufficient chlorine dose. Our results emphasize the capabilities and limitations of the conventional wastewater treatment process in reducing the SARS-CoV-2 RNA concentration and present preliminary evidence for the importance of tertiary treatment and chlorination in reducing dissemination of the virus to the environment., We present data on SARS-CoV-2 RNA concentration and removal rates during the different stages of the activated sludge wastewater treatment process with implications for controlling environmental viral dissemination.

The main objective of this study was to quantify the distribution of pathogens and antibiotic resistance genes in the vadose zone of the soil aquifer treatment (SAT) system. Soil samples were collected from a treated wastewater infiltration basin to a depth of 25 m in two sampling events: (i) at the end of flooding and infiltration and (ii) following three days of drying before the subsequent flooding event. Viable count parallelly of bacteria compared with microscopic live/dead count and enzymatic activity FDA hydrolysis. The abundance of the total bacteria, coliform, antibiotic resistance bacteria (ARB), were examined. In addition, total genomic DNA was extracted from soil samples (n-28 for both flooding and drying cycle), and quantitative PCR (qPCR) was used to determine the relative abundance of antimicrobial resistance genes (ARGs), including 1 integron-integrase intI1, blaTEM, blaCTX-M-32, sul1, qnrS.In both sampling events, the results demonstrate that the distribution of antibiotic resistance genes in the vadose zone exhibits a similar pattern to the one obtained for the examined pathogen. We observed a high concentration of pathogens in topsoil layers and a gradual decline with depth. In this presentation, the profile obtained will be described and discussed with pathogens and ARGs transport and retention in the SAT system.

Multi-elemental C-Br-Cl compound-specific isotope analysis was applied for characterizing abiotic and biotic degradation of the environmental pollutant 1-bromo-2-chloroethane (BCE). Isotope effects were determined in the model processes following hydrolytic dehalogenation and dihaloelimination pathways as well as in a microcosm experiment by the microbial culture from the contaminated site. Hydrolytic dehalogenation of BCE under alkaline conditions and by DhaA enzyme resulted in similar dual isotope slopes (ɅC/Br 21.9 ± 4.7 and 19.4 ± 1.8, respectively, and ɅC/Cl ~ ∞). BCE transformation by cyanocobalamin (B12) and by Sulfurospirillum multivorans followed dihaloelimination and was accompanied by identical, within the uncertainty range, dual isotope slopes (ɅC/Br 8.4 ± 1.7 and 7.9 ± 4.2, respectively, and ɅC/Cl 2.4 ± 0.3 and 1.5 ± 0.6, respectively). Changes over time in the isotope composition of BCE from the contaminated groundwater showed only a slight variation in δ13C values and were not sufficient for the elucidation of the BCE degradation pathway in situ. However, an anaerobic microcosm experiment with the enrichment cultures from the contaminated groundwater presented dual isotope slopes similar to the hydrolytic pathway, suggesting that the potential for BCE degradation in situ by the hydrolytic dehalogenation pathway exists in the contaminated site.

The reuse of household greywater is increasing globally. Wastewater and greywater treatment processes are not fully effective in removing all contaminants, such as emerging micropollutants, antimicrobial-resistant bacteria, and antibiotic resistance genes. The dynamics between emerging micropollutants and antibiotic resistance genes in greywater treatment systems are complex. Thus, this review aims to analyze the current knowledge on sources, spread, and the fate of emerging micropollutants, antibiotic-resistance genes, and antimicrobial-resistant bacteria in microbial communities of greywater and downstream recipients. The fate of antimicrobial resistance and emerging micropollutants from greywater in the environment has not been determined. More studies are needed to identify the mechanism/s involved in the degradation of emerging micropollutants and the presence of transformation pathways in the microbial metagenome. In the review, we aim to describe the link between the persistence of emerging micropollutants and the emergence of antimicrobial resistance. We showed that the effect of irrigation with treated wastewater was variable. In addition, we tried to summarize the impact of emerging micropollutants on bacteria and their fate in the soil microbiome, demonstrating that emerging micropollutants induce changes in the diversity of soil bacteria. The fate and transport of emerging micropollutants, antimicrobial-resistant bacteria, and antibiotic resistance genes can vary with soil properties. It is, therefore, necessary to better understand how widely antibiotic-resistance genes are disseminated.

2,4,6-trinitrotoluene (TNT) is a highly toxic explosive that contaminates soil and water and may interfere with the degradation of co-occurring compounds, such as hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX). We proposed that TNT may influence RDX-degrading bacteria via either general toxicity or a specific effect on the |RDX degradation mechanisms. Thus, we examined the impact of TNT on RDX degradation by Rhodococcus strains YH1, T7, and YY1, which were isolated from an explosives-polluted environment. Although partly degraded, TNT did not support the growth of any of the strains when used as either sole carbon or sole nitrogen sources, or as carbon and nitrogen sources. The incubation of a mixture of TNT (25 mg/l) and RDX (20 mg/l) completely inhibited RDX degradation. The effect of TNT on the cytochrome P450, catalyzing RDX degradation, was tested in a resting cell experiment, proving that TNT inhibits XplA protein activity. A dose-response experiment showed that the IC50/trans values for YH1, T7, and YY1 were 7.272, 5.098, and 9.140 (mg/l of TNT), respectively, illustrating variable sensitivity to TNT among the strains. The expression of xplA was also strongly suppressed by TNT. Cells that were pre-grown with RDX (allowing xplA expression) and incubated with ammonium chloride, glucose, and TNT, completely transformed into their amino dinitrotoluene isomers and formed azoxy toluene isomers. The presence of oxygen-insensitive nitroreductase that enable reduction of the nitro group in the presence of O2 in the genomes of these strains suggests that they are responsible for TNT transformation in the cultures. The experimental results concluded that TNT has an adverse effect on RDX degradation by the examined strains. It inhibits RDX degradation due to the direct impact on cytochrome P450, xplA, or its expression. The tested strains can transform TNT independently of RDX. Thus, degradation of both compounds is possible if TNT concentrations are below their IC50 values.

Biodegradation of pollutants in the environment is directly affected by microbial communities and pollutant mixture at the site. Lab experiments using bacterial consortia and substrate mixtures are required to increase our understanding of these processes in the environment. One of the deficiencies of working with environmental cultures is the inability to culture and identify the active strains while knowing they are representative of the original environment. In the present study, we tested the aerobic microbial degradation of two brominated flame retardants, tribromo-neopentyl alcohol (TBNPA) and dibromo neopentyl glycol (DBNPG), by an assembled bacterial consortium of four strains. The four strains were isolated and plate-cultured from a consortium enriched from the impacted groundwater underlying the Neot Hovav industrial area (Negev, Israel), in which TBNPA and DBNPG are abundant pollutants. Total degradation (3–7 days) occurred only when the four-strain consortium was incubated together (25 °C, pH −7.2) with an additional carbon source, as both compounds were not utilized as such. Bacterial growth was found to be the limiting factor. A dual carbon–bromine isotope analysis was used to corroborate the claim that the isolated strains were responsible for the degradation in the original enriched consortium, thus ensuring that the isolated four-strain microbial consortium is representative of the actual environmental enrichment.

Perchlorate is considered a rapidly spreading environmental pollutant. In Israel, it has been found at high concentrations in the vadose zone (up to 30,000 mg/L) and groundwater (up to 800 mg/L) underlying former industrial waste ponds. A perchlorate-reduction method that utilizes the high degradation potential of shallow soil and the high mobility of perchlorate across the deep unsaturated zone has been proposed. The combined treatment method includes recurrent pumping and application of polluted groundwater amended with an electron donor to the shallow soil layers. As a result, perchlorate is biodegraded in the upper soil, and the treated water drains through the unsaturated zone, displacing the pollutant toward the water table, where it is immediately pumped back to the surface for further treatment through a cyclic process. In the current study, the combined treatment approach was tested in a full-scale unsaturated zone (40 m), long-term (1 year) field experiment. Results showed a daily reduction in perchlorate concentration from 800 mg/L to practically zero. A total of ˜330 kg of perchlorate was reduced during the experiment. Nevertheless, competitive reduction (iron and sulfate) and soil acidification were found to be limiting factors. The study demonstrates a potentially efficient way to overcome these limitations by optimizing electron donor concentration.

Phenyl urea herbicides are being extensively used for weed control in both agricultural and non-agricultural applications. Linuron is one of the key herbicides in this family and is in wide use. Like other phenyl urea herbicides, it is known to have toxic effects as a result of its persistence in the environment. The natural removal of linuron from the environment is mainly carried through microbial biodegradation. Some microorganisms have been reported to mineralize linuron completely and utilize it as a carbon and nitrogen source. Variovorax sp. strain SRS 16 is one of the known efficient degraders with a recently sequenced genome. The genomic data provide an opportunity to use a genome-scale model for improving biodegradation. The aim of our study is the construction of a genome-scale metabolic model following automatic and manual protocols and its application for improving its metabolic potential through iterative simulations. Applying flux balance analysis (FBA), growth and degradation performances of SRS 16 in different media considering the influence of selected supplements (potential carbon and nitrogen sources) were simulated. Outcomes are predictions for the suitable media modification, allowing faster degradation of linuron by SRS 16. Seven metabolites were selected for in vitro validation of the predictions through laboratory experiments confirming the degradation-promoting effect of specific amino acids (glutamine and asparagine) on linuron degradation and SRS 16 growth. Overall, simulations are shown to be efficient in predicting the degradation potential of SRS 16 in the presence of specific supplements. The generated information contributes to the understanding of the biochemistry of linuron degradation and can be further utilized for the development of new cleanup solutions without any genetic manipulation.

Azospirillum brasilense Sp7 and Azospirillum brasilense Cd are two plant growth-promoting bacteria (PGPB). Traditional inoculation methods with PGPB are seed inoculation, seedling root inoculation and soil inoculation. Although these methods are simple to use, they are limited to apply at a specific plant growth stage. Therefore, PGPB inoculation by drip irrigation has been suggested as a means to deliver PGPB directly to the root zone during the plant growth stages. To quantify the intrinsic transport characteristics of two A. brasilense strains following point source inoculation, the properties of A. brasilense Sp7 and A. brasilense Cd (e.g., cell size, hydrophobicity, and zeta potential) and the adsorption characteristics on fine sand were measured. The transport and fate of the two strains were examined under transient water flow conditions with three soil inoculation regimes: (i) surface irrigation (ii) subsurface irrigation and (iii) soil premixing. The water content, bromide, and bacteria distribution in the soil profile were measured after 2 and 48 hours. The measured data were described using the attachment/detachment model using the Hydrus 2/3D code. The result showed that even though A. brasilense Sp7 and Cd exhibit similar hydrophilicity and zeta potential their adsorption and/or straining in the soil profile were differed. A. brasilense Cd has a smaller cell size, less adsorption and less straining than A. brasilense Sp7, thus its vertical movement is deeper. However, both strains accumulated at the vicinity of the water source. The results of this study will be presented and the pros and cons of three inoculation regimes will be discussed.

Complex microbial communities facilitate iron and methane transformations in anoxic methanic sediments of freshwater lakes, such as Lake Kinneret (the Sea of Galilee, Israel). The phylogenetic and functional diversity of these consortia are not fully understood, and it is not clear which lineages perform iron reduction and anaerobic oxidation of methane (AOM). Here, we investigated microbial communities from both natural Lake Kinneret iron-rich methanic sediments (>20 cm depth) and iron-amended slurry incubations from this zone using metagenomics, focusing on functions associated with iron reduction and methane cycling. Analyses of the phylogenetic and functional diversity indicate that consortia of archaea (mainly Bathyarchaeia, Methanomicrobia, Thermoplasmata, and Thermococci) and bacteria (mainly Chloroflexi (Chloroflexota), Nitrospirae (Nitrospirota), and Proteobacteria) perform key metabolic reactions such as amino acid uptake and dissimilation, organic matter fermentation, and methanogenesis. The Deltaproteobacteria, especially Desulfuromondales (Desulfuromonadota), have the potential to transfer electrons extracellularly either to iron mineral particles or to microbial syntrophs, including methanogens. This is likely via transmembrane cytochromes, outer-membrane hexaheme c-type cytochrome (OmcS) in particular, or pilin monomers (PilA), all of which were attributed to this lineage. Bona fide anaerobic oxidizers of methane (ANME) and denitrifying methanotrophs Methylomirabilia (NC10) may mediate AOM in these methanogenic sediments; however we also consider the role of methanogens in active AOM or back flux of methanogenesis. Putative aerobes, such as methane-oxidizing bacteria Methylomonas and their methylotrophic syntrophs Methylotenera, are found among the anaerobic lineages in Lake Kinneret iron-amended slurries and are also involved in the oxidation of methane or its intermediates, as suggested previously. We propose a reaction model for the metabolic interactions in these sediments, linking the potential players that interact via intricate metabolic tradeoffs and direct electron transfer between species. Our results highlight the metabolic complexity of microbial communities in an energy-limited environment, where aerobe and anaerobe communities may co-exist and facilitate AOM as one strategy for survival.

Complex microbial communities facilitate iron and methane transformations in anoxic ‎methanic sediments of freshwater lakes, such as Lake Kinneret (The Sea of Galilee, Israel). The ‎phylogenetic and functional diversity of these consortia are not fully understood, and it is not ‎clear which lineages perform iron reduction, anaerobic oxidation of methane (AOM) or both ‎‎(Fe-AOM). Here, we investigated microbial communities from both natural Lake Kinneret ‎sediments and iron-amended slurry incubations using metagenomics, focusing on functions ‎associated with iron reduction and methane cycling. Analyses of the phylogenetic and ‎functional diversity indicate that consortia of archaea (mainly Bathyarchaeia, ‎Methanomicrobia, Thermoplasmata, and Thermococci) and bacteria (mainly Chloroflexi ‎‎(Chloroflexota), Nitrospirae (Nitrospirota) and Proteobacteria) perform key metabolic ‎reactions such as amino acid uptake and dissimilation, organic matter fermentation, and ‎methanogenesis. The intrinsic Deltaproteobacteria, especially Desulfuromondales ‎‎(Desulfuromonadota), have the potential to transfer electrons extracellularly either to iron ‎mineral particles or to microbial syntrophs, including methanogens. This is likely via ‎transmembrane cytochromes, outer membrane hexaheme c-type cytochrome (OmcS) in ‎particular, or pilin monomer, PilA, which were attributed to this lineage. The bonafide ‎anaerobic oxidizers of methane (ANME) and denitrifying methanotrophs Methylomirabilia ‎‎(NC10) were scarce, and we consider the role of the lineage Methanothrix (Methanothrichales) ‎in Fe-AOM. We show that putative aerobes, such as methane-oxidizing bacteria Methylomonas ‎and their methylotrophic synthrophs Methylotenera are found among the anaerobic lineages in ‎Lake Kinneret iron amended slurries and can be involved in the oxidation of methane or its ‎intermediates, as suggested previously. We propose a reaction model for metabolic interactions ‎in the lake sediments, linking the potential players that interact via intricate metabolic ‎tradeoffs and direct electron transfer between species. Our results highlight the metabolic ‎complexity of microbial communities in an energy-limited environment, where aerobe and ‎anaerobe communities may co-exist and facilitate Fe-AOM as one strategy for survival.‎

We report the draft genome sequences for Rhodococcus sp. strains YH1 and T7. These strains are both capable of degrading hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and were isolated from explosive-contaminated soil and groundwater, respectively. Further genomic analysis might facilitate an understanding of the degradation of RDX and will contribute to the development of bioremediation methods for polluted soil and groundwater.

We report the draft genome sequences for Rhodococcus sp. strains YH1 and T7. These strains are both capable of degrading hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and were isolated from explosive-contaminated soil and groundwater, respectively. Further genomic analysis might facilitate an understanding of the degradation of RDX and will contribute to the development of bioremediation methods for polluted soil and groundwater.

SARS-CoV-2 is an RNA virus, a member of the coronavirus family of respiratory viruses that includes SARS-CoV-1 and MERS. COVID-19, the clinical syndrome caused by SARSCoV-2, has evolved into a global pandemic with more than 2,900,000 people infected. It has had an acute and dramatic impact on health care systems, economies, and societies of affected countries within these few months. Widespread testing and tracing efforts are employed in many countries in order to contain and mitigate this pandemic. Recent data has indicated that fecal shedding of SARS-CoV-2 is common, and that the virus can be detected in wastewater. This indicates that wastewater monitoring is a potentially efficient tool for epidemiological surveillance of SARS-CoV-2 infection in large populations at relevant scales. Collecting raw sewage data, representing specific districts, and crosslinking this data with the number of infected people from each location, will enable us to derive and provide quantitative surveillance tools. In particular, this will provide important means to (i) estimate the extent of outbreaks and their spatial distributions, based primarily on in-sewer measurements (ii) manage the early-warning system quantitatively and efficiently (and similarly, verify disease elimination). Here we report the development of a virus concentration method using PEG or alum, providing an important a tool for detection of SARS-CoV-2 RNA in sewage and relating it to the local populations and geographic information. This will provide a proof of concept for the use of sewage associated virus data as a reliable epidemiological tool.

In disenfranchised communities, untreated greywater (wastewater without sewage) is often environmentally discharged, resulting in potential human exposure to antimicrobial-resistant bacteria (ARB), including extended-spectrum beta-lactamase (ESBL) producers. We sought to examine the abundance of ARB, specifically ESBLs, and antimicrobial resistance genes (ARGs) in greywater from off-grid, pastoral Bedouin villages in Southern Israel. Greywater samples (n = 21) collected from five villages were analyzed to enumerate fecal coliforms and Escherichia coli. ESBL producers were recovered on CHROMagar ESBL and confirmed by VITEK&reg, 2 (bioMerieux, Marcy l'Etoile, France) for identification and antimicrobial susceptibility testing. Total genomic DNA was extracted from greywater samples and quantitative PCR (qPCR) was used to determine relative abundance (gene copies/16S rRNA gene) of class 1 integron-integrase intI1, blaTEM, blaCTX-M-32, sul1, and qnrS. The mean count of presumptive ESBL-producing isolates was 4.5 &times, 106 CFU/100 mL. Of 81 presumptive isolates, 15 ESBL producers were recovered. Phenotypically, 86.7% of ESBL producers were multi-drug resistant. Results from qPCR revealed a high abundance of intI1 (1.4 &times, 10&minus, 1 gene copies/16S rRNA), sul1 (5.2 &times, 2 gene copies/16S rRNA), and qnrS (1.7 &times, 2 gene copies/16S rRNA) followed by blaTEM (3.5 &times, 3 gene copies/16S rRNA) and blaCTX-M-32 (2.2 &times, 5 gene copies/16S rRNA. Results from our study indicate that greywater can be a source of ARB, including ESBL producers, in settings characterized by low sanitary conditions and inadequate wastewater management.

Membrane biofouling constitutes a great challenge in anaerobic membrane bioreactor (AnMBR). Here, we studied the initial deposition of anaerobes, the first step in biofilm formation, with a consortium isolated from an AnMBR on membranes with different surface properties and under two shear rate conditions without filtration. We found that the cell transfer coefficient, calculated from the initial deposition experiments, was similar under the two shear rates for the hydrophobic membranes, but much higher under low shear rate and much lower under high shear rate, for the hydrophilic membrane. The cell transfer coefficient measured under filtration mode and at a higher shear rate showed a similar trend. The pioneer bacteria and archaea (without filtration) were identified by next-generation sequencing. The results showed that the selective force for the dissimilarity of the pioneer bacterial and archaeal diversity was the shear rate and the membrane surface properties, respectively. However, statistical analyses revealed minor changes in the pioneer bacteria (class level) and archaea (order level) populations under the various conditions. These results shed light on the first step of biofilm formation on the membranes in AnMBRs and emphasize the importance of hydrodynamic shear and membrane surface properties on the initially deposited anaerobes.

We report the whole-genome sequence of Gordonia sp. strain YY1, which was isolated from the surface soil in an explosive-contaminated site in Israel and cultivated with hexahydro-1,3,5-trinitro-1,3,5-triazine, i.e., royal demolition explosive (RDX), as a nitrogen source. This genome sequence will improve our understanding of the genes for RDX degradation. In addition, this research will reveal metabolic pathways in order to develop new bioremediation methods for polluted soil and groundwater.