Role of endogenous soil microorganisms in controlling antimicrobial resistance after the exposure to treated wastewater.

The reuse of treated wastewater (TWW) for irrigation appears to be a relevant solution to the challenges of growing water demand and scarcity. However, TWW contains not only micro-pollutants including pharmaceutical residues but also antibiotic resistant bacteria. The reuse of TWW could contribute to the dissemination of antimicrobial resistance in the environment. The purpose of this study was to assess if exogenous bacteria from irrigation waters (TWW or tap water-TP) affect endogenous soil microbial communities (from 2 soils with distinct irrigation history) and key antibiotic resistance gene sul1 and mobile genetic elements intl1 and IS613. Experiments were conducted in microcosms, irrigated in one-shot, and monitored for three months. Results showed that TP or TWW exposure induced a dynamic response of soil microbial communities but with no significant increase of resistance and mobile gene abundances. However, no significant differences were observed between the two water types in the current experimental design. Despite this, the 16S rDNA analysis of the two soils irrigated for two years either with tap water or TWW resulted in soil microbial community differentiation and the identification of biomarkers from Xanthomonadaceae and Planctomycetes families for soils irrigated with TWW. Low-diversity soils were more sensitive to the addition of TWW. Indeed, TWW exposure stimulated the growth of bacterial genera known to be pathogenic, correlating with a sharp increase in the copy number of selected resistance genes (up to 3 logs). These low-diversity soils could thus enable the establishment of exogenous bacteria from TWW which was not observed with native soils. In particular, the emergence of Planctomyces, previously suggested as a biomarker of soil irrigated by TWW, was here demonstrated. Finally, this study showed that water input frequency, initial soil microbial diversity and soil history drive changes within soil endogenous communities and the antibiotic resistance gene pool.

Fate of nitrogen and phosphorus from source-separated human urine in a calcareous soil.

Human urine concentrates 88% of the nitrogen and 50% of the phosphorus excreted by humans, making it a potential alternative crop fertilizer. However, knowledge gaps remain on the fate of nitrogen in situations favouring NH3 volatilization and on the availability of P from urine in soils. This study aimed at identifying the fate of nitrogen and phosphorus supplied by human urine from source separation toilets in a calcareous soil. To this end, a spinach crop was fertilized with 2 different doses of human urine (170 kgN ha-1 + 8.5 kgP ha-1 and 510 kgN ha-1 + 25.5 kgP ha-1) and compared with a synthetic fertilizer treatment (170 kgN ha-1 + 8.5 kgP ha-1) and an unfertilized control. The experiment was conducted in 4 soil tanks (50-cm depth) in greenhouse conditions, according to a randomized block scheme. We monitored soil mineral nitrogen over time and simulated nitrogen volatilization using Hydrus-1D and Visual Minteq softwares. We also monitored soil phosphorus pools, carbon, nitrogen and phosphorus (CNP) in microbial biomass, soil pH and electrical conductivity. Only an excessive input of urine affected soil pH (decreasing it by 0.2 units) and soil conductivity (increasing it by 183%). The phosphorus supplied was either taken up by the crop or remained mostly in the available P pool, as demonstrated by a net increase of the resin and bicarbonate extractable P. Ammonium seemed to be nitrified within about 10 days after application. However, both Visual Minteq and Hydrus models estimated that more than 50% of the nitrogen supplied was lost by ammonia volatilization. Overall, our results indicate that direct application of urine to a calcareous soil provides available nutrients for plant growth, but that heavy losses of volatilized nitrogen are to be expected. Our results also question whether long-term application could affect soil pH and salinity.

Clustering and mapping the first COVID-19 outbreak in France.

BACKGROUND: With more than 160 000 confirmed COVID-19 cases and about 30 000 deceased people at the end of June 2020, France was one of the countries most affected by the coronavirus crisis worldwide. We aim to assess the efficiency of global lockdown policy in limiting spatial contamination through an in-depth reanalysis of spatial statistics in France during the first lockdown and immediate post-lockdown phases. METHODS: To reach that goal, we use an integrated approach at the crossroads of geography, spatial epidemiology, and public health science. To eliminate any ambiguity relevant to the scope of the study, attention focused at first on data quality assessment. The data used originate from official databases (Santé Publique France) and the analysis is performed at a departmental level. We then developed spatial autocorrelation analysis, thematic mapping, hot spot analysis, and multivariate clustering. RESULTS: We observe the extreme heterogeneity of local situations and demonstrate that clustering and intensity are decorrelated indicators. Thematic mapping allows us to identify five "ghost" clusters, whereas hot spot analysis detects two positive and two negative clusters. Our re-evaluation also highlights that spatial dissemination follows a twofold logic, zonal contiguity and linear development, thus determining a "metastatic" propagation pattern. CONCLUSIONS: One of the most problematic issues about COVID-19 management by the authorities is the limited capacity to identify hot spots. Clustering of epidemic events is often biased because of inappropriate data quality assessment and algorithms eliminating statistical-spatial outliers. Enhanced detection techniques allow for a better identification of hot and cold spots, which may lead to more effective political decisions during epidemic outbreaks.

Life cycle assessment as decision support tool for water reuse in agriculture irrigation.

This study presents a decision support tool that evaluates the environmental efficiency of water reclamation for agricultural irrigation, among other options. The developed tool is published as open source at https://doi.org/10.18167/DVN1/YLP1BA. The objective of this decision support tool is to facilitate the interpretation of the Life Cycle Assessment (LCA) results. This framework was applied to a representative case of reuse of reclaimed water for vine irrigation at the Murviel-Les-Montpellier experimental site (Hérault, France). It was then generalized through modeling assumptions to consider different reuse scenarios. To highlight situations in which the supply of recycled water for irrigation may or may not provide significant environmental benefits, three main parameters were varied: (i) tertiary treatment technologies, (ii) availability of conventional water sources, (iii) energy mix composition. The results show that the environmental impact of reclaimed water depends directly on the type of tertiary treatment technology and the location of the treatment plant in relation to the field and other water sources. The decision support tool has identified where wastewater reuse is clearly an environmentally beneficial source of irrigation among surface and groundwater sources (e.g., WWTP closer to field than river, groundwater too deep, tertiary treatment environmentally beneficial). However, there are many situations where the decision support process cannot distinguish between water reuse for agricultural irrigation and conventional water sources, especially when the nutrient content of treated municipal wastewater is insufficient to offset the negative effects of high energy requirements and chemicals of tertiary treatment.

Treated wastewater reuse in micro-irrigation: effect of shear stress on biofilm development kinetics and chemical precipitation.

Treated wastewater in micro-irrigation is a promising approach that could be used to decrease the pressure on good quality water resources. However, the clogging of such systems due to biofilm development and chemical precipitation constitute a constraint with the use of treated wastewater (TWW) and lead to lower irrigation system performance. The objective of this work is to study the development of biofilm and composition of fouling due to TWW under shear stresses of 0.7, 2.2 and 4.4 Pa detected along micro-irrigation systems. For this purpose, a Taylor-Couette reactor (TCR) was specifically calibrated for the cultivation of biofilm. The analysis of fouling composition samples (organic and inorganic) shows that biofilm tends to develop under the highest shear stress value (4.4 Pa). Precipitation of calcium carbonate in the form of calcite was observed in conjunction with biofilm growth using X-ray diffractometry (XRD) and thermogravimetric analysis (TGA). These results can be used to ascertain the origins of chemical and biological clogging of drippers and fouling of pipes related to reclaimed water- irrigation.

Impact of long-term irrigation with municipal reclaimed wastewater on the uptake and degradation of organic contaminants in lettuce and leek.

A two years drip irrigation of lettuce and leek crops with treated municipal wastewater without and with spiking with fourteen wastewater relevant contaminants at 10 µg/L concentration level was conducted under greenhouse cultivation conditions to investigate their potential accumulation in soil and leaves and to assess human health related risks. Lettuce and leek crops were selected as a worse-case scenario since leafy green vegetable has a high potential for organic contaminants uptake. The results revealed limited accumulation of contaminants in soil and plant leaves, their concentration levels being in the range of 1-30 ng/g and 1-660 ng/g range in soil and leaves, respectively. This was likely related to abiotic and biotic transformation or simply binding processes in soil, which limited contaminants plant uptake. This assumption was underpinned by studies of the enantiomeric fractionation of chiral compounds (e.g. climbazole and metoprolol) in soil as pieces of evidence of biodegradation and by the identification of transformation products or metabolites in leaves by means of liquid chromatography - high resolution - mass spectrometry using a suspect screening workflow. The high bioconcentration factors were not limited to compounds with intermediate Dow (100 to 1000) such as carbamazepine but also observed for hydrophilic compounds such as clarithromycin, hydrochlorothiazide and the food additives acesulfame and sucralose. This result assumed that accumulation was not only driven by passive processes (e.g. lipoidal diffusion through lipid bilayer cell membranes or Casparian strip) but might be supported by carrier-mediated transporters. As a whole, this study confirmed earlier reports on the a de minimis human health risk related to the consumption of raw leafy green vegetable irrigated with domestic TWW containing organic contaminants residues.

Effects of the chlorination and pressure flushing of drippers fed by reclaimed wastewater on biofouling.

Milli-channel baffle labyrinths are widely used in drip irrigation systems. They induce a pressure drop enabling drip irrigation. However, with a section thickness that is measured in mm2, they are sensitive to clogging, which reduces the performance and service life of a drip irrigation system. The impact of chlorination (1.5 ppm of free chlorine during 1 h application) and pressure flushing (0.18 MPa) on the biofouling of non-pressure-compensating drippers, fed by real reclaimed wastewater, was studied at lab scale using optical coherence tomography. The effect of these treatments on microbial composition (bacteria and eukaryotes) was also investigated by High-throughput DNA sequencing. Biofouling was mainly observed in the inlet, outlet and return areas of the milli-labyrinth channel from drippers. Chlorination reduced biofilm development, particularly in the mainstream of the milli-labyrinth channel, and it was more efficient when combined with pressure flushing. Moreover, chlorination was more efficient in maintaining water distribution uniformity (CU < 95% compared to less than 85% for unchlorinated lines). It reduced more efficiently the bacterial concentration (≈1 log) and the diversity of the bacterial community in the dripper biofilms compared to the pressure flushing method. Chlorination significantly modified the microbial communities, promoting chlorine-resistant bacteria such as Comamonadaceae or Azospira. Inversely, several bacterial groups were identified as sensitive to chlorination such as Chloroflexi and Planctomycetes. Nevertheless, one month after stopping the treatments bacterial diversity recovered and the chlorine-sensitive bacteria such as Chloroflexi phylum and the Saprospiraceae, Spirochaetaceae, Christensenellaceae and Hydrogenophilaceae families re-emerged in conjunction with the growth of biofouling, highlighting the resilience of the bacteria originating from drippers. Based on PCoA analyses, the structure of the bacterial communities still clustered separately from non-chlorinated drippers, showing that the effect of chlorination was still detectable one month after stopping the treatment.

Post-lockdown detection of SARS-CoV-2 RNA in the wastewater of Montpellier, France.

The evolution of the COVID-19 pandemic can be monitored through the detection of SARS-CoV-2 RNA in sewage. Here, we measured the amount of SARS-CoV-2 RNA at the inflow point of the main waste water treatment plant (WWTP) of Montpellier, France. We collected samples 4 days before the end of lockdown and up to 70 days post-lockdown. We detected increased amounts of SARS-CoV-2 RNA at the WWTP from mid-June on, whereas the number of new COVID-19 cases in the area started increasing a couple of weeks later. Future epidemiologic investigations shall explain such asynchronous finding.

Hydrodynamic effect on biofouling of milli-labyrinth channel and bacterial communities in drip irrigation systems fed with reclaimed wastewater.

The clogging of drippers due to the development of biofilms reduces the benefits and is an obstacle to the implementation of drip irrigation technology in a reclaimed water context. The narrow section and labyrinth geometry of the dripper channel results the development of a heterogeneous flow behaviours with the vortex zones which it enhance the fouling mechanisms. The objective of this study was to analyse the influence of the three dripper types, defined by their geometric and hydraulic parameters, fed with reclaimed wastewater, on the biofouling kinetics and the bacterial communities. Using optical coherence tomography, we demonstrated that the inlet of the drippers (mainly the first baffle) and vortex zones are the most sensitive area for biofouling. Drippers with the lowest Reynolds number and average cross-section velocity v (1 l·h-1) were the most sensible to biofouling, even if detachment events seemed more frequent in this dripper type. Therefore, dripper flow path with larger v should be consider to improve the anti-clogging performance. In addition, the dripper type and the geometry of the flow path influenced the structure of the bacterial communities from dripper biofilms. Relative abundancy of filamentous bacteria belonging to Chloroflexi phylum was higher in 1 l·h-1 drippers, which presented a higher level of biofouling. However, further research on the role of this phylum in dripper biofouling is required.

Drip irrigation biofouling with treated wastewater: bacterial selection revealed by high-throughput sequencing.

Clogging of drippers due to the development of biofilms weakens the advantages and impedes the implementation of drip irrigation technology. The objective of this study was to characterise the bacterial community of biofilms that develop in a drip irrigation system supplied with treated wastewater. High-throughput sequencing of 16S rRNA gene amplicons indicated that the bacterial community composition differed between drippers and pipes, mainly due to changes in the abundance of the genus Aquabacterium. Cyanobacteria were found to be involved in the biological fouling of drippers. Moreover, bacterial genera including opportunistic pathogenic bacteria such as Legionella and Pseudomonas were more abundant in dripper and pipe biofilms than in the incoming water. Some genera such as Pseudomonas were mostly recovered from drippers, while others (ie Bacillus, Brevundimonas) mainly occurred in pipes. Variations in the hydraulic conditions and properties of the materials likely explain the shift in bacterial communities observed between pipes and drippers.

Methodology to Evaluate Dripper Sensitivity to Clogging due to Solid Particles: An Assessment.

Emitter clogging is a major problem in microirrigation systems, which may result from the isolated or combined effects of physical, chemical, and biological agents. Clogging caused by suspended solid particles is the most common plugging form of emitters. Water quality and emitter geometry are key aspects in clogging processes. Any suitable test procedure to assess the sensitivity of drippers to clogging should take into account the predominant factors that influence clogging and must reproduce the field conditions. This research set out to assess the performance and suitability of a laboratory clogging test procedure in order to validate a methodology and to provide scientific results that may support the standardization of a clogging test method. The evaluated methodology has been used by the IRSTEA laboratory since 1974 (Platform of Research and experiment on Science and Technology for Irrigation - PReSTI, formerly LERMI) and its contents are currently being discussed by the ISO TC23/SC18 committee. The aim is to define a standardized testing protocol to evaluate the sensitivity of emitters to clogging due to solid particles. Replications analyzing the clogging resistance of four models of emitting pipes were carried out in a laboratory. The clogging test procedure enabled an accurate assessment of the combinations of concentration and size of particles that caused clogging in each model of dripper. However, a significant variability in degree of clogging was identified when the results of replications for each model of dripper were compared. Several requirements, concerns, and improvements related to the clogging test protocol were discussed.