Evolution of microbiome composition, antibiotic resistance gene loads, and nitrification during the on-farm composting of the solid fraction of pig slurry using two bulking agents.

Composting is a nature-based method used to stabilize organic matter and to transform nitrogen from animal farm manure or solid fraction of slurry (SFS). The use of composted material as source of nutrients for agriculture is limited by its potential to facilitate the propagation of biological hazards like pathogens and antibiotic-resistant bacteria and their associated antibiotic-resistance genes (ARG). We show here an experimental on-farm composting (one single batch) of pig SFS, performed under realistic conditions (under dry continental Mediterranean climate) for 280 days, and using two different bulking agents (maize straw and tree pruning residues) for the initial mixtures. The observed reduction in potentially pathogenic bacteria (80-90%) and of ARG loads (60-100%) appeared to be linked to variations in the microbiome composition occurring during the first 4 months of composting, and concurrent with the reduction of water-soluble ammonium and organic matter loads. Nitrification during the composting has also been observed for both composting piles. Similar patterns have been demonstrated at small scale and the present study stresses the fact that the removal can also occur at full scale. The results suggest that adequate composition of the starting material may accelerate the composting process and improve its global performance. While the results confirm the sanitization potential of composting, they also issue a warning to limit ARG loads in soils and in animal and human gut microbiomes, as the only way to limit their presence in foodstuffs and, therefore, to reduce consumers' exposure.

Removal and environmental risk assessment of contaminants of emerging concern from irrigation waters in a semi-closed microalgae photobioreactor.

The present study evaluated the efficiency of a semi-closed, tubular, horizontal photobioreactor (PBR) to treat a mixture of irrigation and rural drainage water, focusing in the removal of different contaminants of emerging concern (CECs), and evaluating the environmental impact of the resulting effluent. Target CECs included pharmaceuticals, personal care products and flame retardants. Of the 13 compounds evaluated, 11 were detected in the feed water entering the PBR, and diclofenac (DCF) (1107 ng L-1) and N,N-diethyl-toluamide (DEET) (699 ng L-1) were those present at the greatest concentrations. The best removal efficiencies were achieved for the pharmaceuticals diazepam (94%), lorazepam (LZP) (83%) and oxazepam (OXA) (71%), and also for ibuprofen (IBU) (70%). For the rest of the CECs evaluated, attenuation was similar to that obtained after conventional wastewater treatment, ranging from basically no elimination (carbamazepine (CBZ) and tris-(2-chloroethyl) phosphate (TCEP)) to medium efficiencies (DCF and tributyl phosphate (TBP) (50%)). Environmental risk assessment based on hazard quotients (HQs) resulted in HQ values < 0.1 (no risk associated) for most of the compounds and most of the trophic levels considered. Values between 1 and 10 (moderate risk) were obtained for tonalide (AHTN) (fish) and CBZ (invertebrates). The most sensitive trophic level was green algae, whereas fish and aquatic plants were the most resilient. Our results suggest that microalgae-based treatments could become a green, cost-effective alternative to conventional wastewater treatment regarding the efficient elimination of these contaminants.

Does the application of human waste as a fertilization material in agricultural production pose adverse effects on human health attributable to contaminants of emerging concern?

The QuEChERS (quick, easy, cheap, effective, rugged, and safe) ultrasound-assisted method was successfully used to determine the presence of contaminants of emerging concern (CECs) in both the growing medium (i.e. soil) and vegetable samples (i.e. potato, onion, celery, parsnip and carrot) cultivated in household gardens under field conditions impacted by the application of livestock manure mixed with human waste. CECs with a broad range of physico-chemical properties including pharmaceuticals, plasticizers, herbicides, personal care products, and biocides were investigated. Among all studied CECs, diclofenac (DCL), carbamazepine, bisphenol A (BPA) and estrogenic hormones (estrone (E1), 17α-ethinyl estradiol (EE2)) were detected and quantified in vegetable samples with a concentration range from 0.114 to 13.3 ng/g fresh weight (f.w.), while BPA, E1, and EE2 were detected in soil samples with concentration ranges from 0.526 to 0.830 ng/g f.w., 0.121-0.199 ng/g f.w. and 0.118-0.333 ng/g f.w., respectively. DCL was only quantified above the limit of quantification in one soil sample (0.151 ng/g f.w.). The human health risk was estimated using the threshold of toxicological concern approach and a standard diet for two segments of the population (i.e. adult and child). The consumption of investigated vegetables contaminated with CECs poses no risk for the Serbian population.

Occurrence and human health risk assessment of antibiotics and trace elements in Lactuca sativa amended with different organic fertilizers.

Soil amendment with organic fertilizers is an effective approach to improve soil fertility. However, organic fertilizers may contain pollutants such as trace elements (TEs) and antibiotics (ABs), which, once deployed in arable soil, can be taken up by vegetables and have adverse effects on crops and human health. This study assesses the presence of 15 TEs and 16 ABs in lettuce grown in a greenhouse facility and amended with 3 different organic fertilizers (sewage sludge (SS), organic fraction of municipal solid waste (OFMSW), and swine manure (SM)) at 3 different fertilization doses. The results show that lettuces amended with SS resulted in the lowest content of TEs. Although 11 ABs were detected in the SM and SS fertilizers, only 3 ABs were detected in lettuce leaves. The concentrations of detected ABs in lettuce ranged from 0.67 ng/g fw (lincomycin) to 14.2 ng/g fw (ciprofloxacin) in SS. The organic fertilization dose did not affect the lettuce uptake of TEs or ABs. Moreover, the use of SS resulted in the highest lettuce yield of the organic amendments. The total hazard quotients (THQs) obtained for TEs and ABs were less than 1 for all the studied fertilization treatments. The highest THQs for TEs were observed in lettuce amended with SM (0.11-0.16), whereas the highest THQs for ABs were observed in SS treatments (0.06-0.09). The results thus suggest that consumption of lettuces amended with organic fertilizers would not pose a risk to human health due to the presence of studied TEs or ABs, but potentially harmful combined effects cannot ruled out.

Dose effect of Zn and Cu in sludge-amended soils on vegetable uptake of trace elements, antibiotics, and antibiotic resistance genes: Human health implications.

The application of sewage sludge to agricultural fields reduces the need for mineral fertilizers by increasing soil organic matter, but may also increase soil pollution. Previous studies indicate that zinc and copper, as the most abundant elements in sewage sludge, affect plant uptake of other contaminants. This paper aims to investigate and compare the effect of increasing amounts of Zn and Cu in sludge-amended soils on the accumulation of trace elements (TEs), antibiotics (ABs), and antibiotic resistance genes (ARGs) in lettuce and radish. The vegetables were grown under controlled conditions, and the influence on plant physiology and human health were also evaluated. The results show that the addition of Zn and Cu significantly increased the concentration of TEs in the edible tissue of both vegetables. According to the hazard quotient (HQ) of the TEs, the human health risk increased 2 to 3 times and was 3-4 times greater in lettuce than in radish. In contrast to the TEs, the occurrence of ABs and most of the ARGs was higher in radish roots than lettuce leaves. ABs were not detected in lettuce leaves, and the amount of all ARGs except blaTEM was 10 times lower than in radish roots. On the other hand, the addition of Zn and Cu had no significant effect on the occurrence of ABs and ARGs in the edible part of the vegetables, and no damage was found to plant productivity or physiology. The results show that the consumption of lettuce and radish grown in sewage-sludge-amended soils under tested doses of Cu and Zn does not pose an adverse human health effect, as the total HQ value was always less than 1, and the presence of ABs and ARGs was not found to have any potential impact. Nevertheless, further studies are needed to estimate the long-term effect on human health of crops grown under frequent application of biosolids in arable soil.

Effects of industrial pollution on the reproductive biology of Squalius laietanus (Actinopterygii, Cyprinidae) in a Mediterranean stream (NE Iberian Peninsula).

Mediterranean rivers are severely affected by pollutants from industry, agriculture and urban activities. In this study, we examined how industrial pollutants, many of them known to act as endocrine disruptors (EDCs), could disturb the reproduction of the Catalan chub (Squalius laietanus). The survey was conducted throughout the reproductive period of S. laietanus (from March to July 2014) downstream an industrial WWTP located in the River Ripoll (NE Iberian Peninsula). Eighty fish (28 females and 52 males) were caught by electrofishing upstream and 77 fish (33 females and 44 males) downstream a WWTP. For both sexes, the gonadosomatic index (GSI) and gonadal histology were examined and related to water chemical analysis and fish biomarkers. Female fecundity was assessed using the gravimetric method. Fish from the polluted site showed enhanced biomarker responses involved in detoxification. Also, in the polluted site, lower GSI values were attained in both sexes and females displayed lower numbers of vitellogenic oocytes. Gonadal histology showed that all maturation stages of testicles and ovaries were present at the two study sites but fish males from the polluted site had smaller diameter seminiferous tubules. Water chemical analysis confirmed greater presence of EDCs in the river downstream the industrial WWTP. The chemicals benzotriazole and benzothiazole could be partially responsible for the observed alterations in the reproductive biology of S. laietanus.

Simultaneous determination of multiclass antibiotics and their metabolites in four types of field-grown vegetables.

The developed method was evaluated for the determination of 10 antibiotics belonging to four chemical classes (fluoroquinolones, sulfonamides, lincosamides, and metoxybenzylpyrimidines) and six of their metabolites in four vegetable matrices (lettuce, tomato, cauliflower, and broad beans). The reported method detection limits were sufficiently low (0.1-5.8 ng/g dry weight) to detect target compounds in vegetables under real agricultural practices. Absolute and relative recovery values ranged from 40 to 118% and from 70 to 118%, respectively, for all targeted compounds at the spike level of 100 ng/g dry weight. Regarding method precision, the highest relative standard deviation (RSD) was obtained for enrofloxacin in lettuce (20%), while for the rest of the compounds in all matrices, the RSD values were below 20% for the same spike level. Matrix effects, due to electrospray ionization, ranged from - 26 to 29% for 85% of all estimated values. In a field study, four of the 10 targeted antibiotics were detected in tested vegetables. For the first time, antibiotic metabolites were quantified in vegetables grown under real field conditions. More specifically, decarboxyl ofloxacin and TMP304 were detected in tomato fruits (1.5 ng/g dry weight) and lettuce leaves (21.0-23.1 ng/g dry weight), respectively. It is important to remark that the concentration of TMP304 was five times higher than that from the parental compound, emphasizing the importance of metabolite analysis in monitoring studies. Therefore, the method provided a robust, reliable, and simple-to-use tool that could prove useful for routine multiclass analysis of antibiotics and their metabolites in vegetable samples. Graphical abstract.

Distribution of antibiotic resistance genes in soils and crops. A field study in legume plants (Vicia faba L.) grown under different watering regimes.

Social concern has raised during the last years due to the development of antibiotic resistance hotspots in different environmental compartments, including the edible parts of crops. To assess the influence of the water quality used for watering, we collected samples from soil, roots, leaves and beans from the legume plant Vicia faba (broad beans) in three agricultural peri-urban plots (Barcelona, NE Spain), irrigated with either groundwater, river water, or reclaimed water. Antibiotic resistance genes (ARGs) sul1, tetM, qnrS1, blaCTX-M-32,blaOXA-58, mecA, and blaTEM were quantified by real-time PCR, along with 16S rDNA and intl1 sequences, as proxies for bacterial abundance and integron prevalence, respectively. Microbiome composition of all samples were analyzed by high-throughput DNA sequencing. Results show a gradient of bacterial species diversity and of ARG prevalence from highly diverse soil samples to microbially-poor beans and leaves, in which Rhizobiales essentially displaced all other groups, and that presented very small loads of ARGs and integron sequences. The data suggest that the microbiome and the associated resistome were likely influenced by agricultural practices and water quality, and that future irrigation water legal standards should consider the specific Physiology of the different crop plants.

Antibiotic resistance gene distribution in agricultural fields and crops. A soil-to-food analysis.

Despite the social concern about the generalization of antibiotic resistance hotspots worldwide, very little is known about the contribution of different potential sources to the global risk. Here we present a quantitative analysis of the distribution of Antibiotic Resistance Genes (ARGs) in soil, rhizospheric soil, roots, leaves and beans in tomato, lettuce and broad beans crops (165 samples in total), grown in nine commercial plots distributed in four geographical zones in the vicinity of Barcelona (North East Spain). We also analyzed five soil samples from a nearby forest, with no record of agricultural activities. DNA samples were analyzed for their content in the ARGs sul1, tetM, qnrS1, blaCTX-M-32, blaOXA-58, mecA, and blaTEM, plus the integron intI1, using qPCR methods. In addition, soil microbiomes from the different plots were analyzed by amplicon-targeted 16S rRNA gene sequencing. Our data show a decreasing gradient of ARG loads from soil to fruits and beans, the latter showing only from 0.1 to 0.01% of the abundance values in soil. The type of crop was the main determinant for both ARG distribution and microbiome composition among the different plots, with minor contributions of geographic location and irrigation water source. We propose that soil amendment and/or fertilization, more than irrigation water, are the main drivers of ARG loads on the edible parts of the crop, and that they should therefore be specifically controlled.

Linking the use of reclaimed water to indicators of crop stress by metabolomic and transcriptomic analyses. A tool to compare water irrigation quality.

The occurrence of contaminants of emerging concern (CECs) or heavy metals in reclaimed water used for agricultural irrigation may affect crop morphology and physiology. Here, we analyzed lettuce (Lactuca sativa) grown in outdoor lysimeters and irrigated with either tap water, used as a control, or reclaimed water: CAS-reclaimed water, an effluent from a conventional activated sludge system (CAS) followed by chlorination and sand filtration, or MBR-reclaimed water, an effluent from a membrane biological reactor (MBR). Chemical analyses identified seven CECs in the reclaimed waters, but only two of them were detected in lettuce (carbamazepine and azithromycin). Metabolomic and transcriptomic analyses revealed that irrigation with reclaimed water increased the concentrations of several crop metabolites (5-oxoproline, leucine, isoleucine, and fumarate) and of transcripts codifying for the plant stress-related genes Heat-Shock Protein 70 (HSP70) and Manganese Superoxide Dismutase (MnSOD). In both cases, MBR-water elicited the strongest response in lettuce, perhaps related to its comparatively high sodium adsorption ratio (4.5), rather than to its content in CECs or heavy metals. Our study indicates that crop metabolomic and transcriptomic profiles depend on the composition of irrigating water and that they could be used for testing the impact of water quality in agriculture.

Potential of nature-based solutions to reduce antibiotics, antimicrobial resistance, and pathogens in aquatic ecosystems. a critical review.

This comprehensive scientific review evaluates the effectiveness of nature-based solutions (NBS) in reducing antibiotics (ABs), combating antimicrobial resistance (AMR), and controlling pathogens in various aquatic environments at different river catchment levels. It covers conventional and innovative treatment wetland configurations for wastewater treatment to reduce pollutant discharge into the aquatic ecosystems as well as exploring how river restoration and saltmarshes can enhance pollutant removal. Through the analysis of experimental studies and case examples, the review shows NBS's potential for providing sustainable and cost-effective solutions to improve the health of aquatic ecosystems. It also evaluates the use of diagnostic indicators to predict NBS effectiveness in removing specific pollutants such as ABs and AMR. The review concludes that NBS are feasible for addressing the new challenges stemming from human activities such as the presence of ABs, AMR and pathogens, contributing to a better understanding of NBS, highlighting success stories, addressing knowledge gaps, and providing recommendations for future research and implementation.

Evaluation of a coagulation/flocculation-lamellar clarifier and filtration-UV-chlorination reactor for removing emerging contaminants at full-scale wastewater treatment plants in Spain.

The presence and elimination of 25 emerging contaminants in two full-scale Spanish wastewater treatment plants was studied. The tertiary treatment systems consisted of coagulation, flocculation lamellar settlement and filtration (pulsed-bed sand filters) units, and disinfection was carried out by medium pressure UV light lamps and chlorination. Diclofenac and carbamazepine were found to be the emerging contaminants with the highest concentrations in secondary effluents. Photodegradable emerging contaminants (e.g. ketoprofen, triclosan and diclofenac) were removed by filtration-UV light radiation-chlorination whereas most hydrophobic compounds (e.g. galaxolide and tonalide) were eliminated by coagulation-flocculation followed by lamellar clarification, a unit in which a seasonal trend was observed. Overall mass removal efficiency was about 60%. 1-(8-Chlorocarbazolyl) acetic acid, an intermediate product of the photodegradation of diclofenac, was detected after filtration-UV-chlorination, but not after coagulation-flocculation and lamellar clarification. This study demonstrated potential for general applicability of two established tertiary treatment systems to eliminate emerging contaminants.

Atmospheric influence on the distribution of organic pollutants in the Guadalquivir River estuary, SW Spain.

In the lower Guadalquivir river basin, a system stressed by a wide variety of anthropogenic activities, eight pesticides (four triazines, two chloroacetanilide herbicides, one organochlorine, and one organophosphorus insecticide); and four emerging pollutants (two personal care products, one organophosphorous flame retardant, and one xanthine alkaloid) were analyzed in river water during a 2-year monitoring program, and after rain episodes. Samples were extracted using the solid phase extraction (SPE) technique prior to determination of compounds using gas chromatograph coupled to a mass spectrometer detector. Except for caffeine, recoveries were mostly above 80 %, while limits of detection and quantification were in the low nanograms per liter level (except for dimethoate). Terbuthylazine, simazine (triazine herbicides), and dimethoate (organophosphorus insecticide), present in agrochemicals, were predominant in the river water, although concentrations were below the quality standards established by the EU Water-Framework-Directive. A general trend to increase concentration was observed after rain events, in particular for pesticides, possibly as a consequence of surface runoff.

Attenuation and soil biodegradation of fungicides by using vegetated buffer strips in vineyards during a simulated rainfall-runoff event.

Rainfall-runoff events occurring in vineyard fields can result in pesticide ground losses and the subsequent pollution of surface water bodies, derivate from the crop protection spray applications. In this study, the capacity of vegetated buffer strips (BS) to prevent surface water pollution due to the application of five fungicide products typically used in vineyards (copper, dimethomorph, oxathiapiprolin, zoxamide, acibenzolar-s-methyl, and laminarin) following a simulated run-off event has been assessed, and compared to that from a bare ground soil (BG). Two strips (5 m in length, each), one with vegetation and the other without were built up, and two different experiments were performed, a runoff event and a soil fungicide degradation kinetic evaluation. The runoff results show that fungicide mass retention in the strips ranged from 73 to 98% and that the presence of vegetation in BS increased the fungicide mass retention in the strips by almost 10% (on average) in comparison to the unvegetated strip. Moreover, soil degradation studies highlighted that the presence of vegetation reduces significantly the half-time life of almost all the studied fungicides by 55%, on average. Eight fungicide transformation products (TPs) were identified following a runoff event in the soil strips, but the abundance of these TPs was up to 78% lower in vegetated strips. These results highlight the effectiveness of using vegetated buffer zones in vineyards to protect aquatic ecosystem pollution.

Phycobiliprotein recovery coupled to the tertiary treatment of wastewater in semi-continuous photobioreactors. Tracking contaminants of emerging concern.

This study evaluated a tertiary wastewater treatment technology using cyanobacteria to recover value-added phycobiliproteins. The presence of contaminants of emerging concern (CECs) in wastewater, cyanobacterial biomass and pigments recovered were also analyzed. For this, a wastewater-borne cyanobacterium (Synechocystis sp. R2020) was used to treat secondary effluent from a municipal wastewater treatment plant, with and without nutrients supplementation. Then, the stability of phycobiliprotein production was assessed by operating the photobioreactor in semi-continuous mode. Results showed similar biomass productivity with and without nutrients supplementation (153.5 and 146.7 mg L-1 d-1, respectively). Upon semi-continuous operation, the phycobiliprotein content was stable and reached up to 74.7 mg gDW-1. The phycocyanin purity ratio ranged from 0.5 to 0.8, corresponding to food grade (>0.7). Out of 22 CECs detected in secondary effluent, only 3 were present in the phycobiliprotein extracts. In order to identify applications, prospective research should focus on CECs removal during pigment purification.

Use of wood and cork in biofilters for the simultaneous removal of nitrates and pesticides from groundwater.

About 13% and 7% of monitored groundwater stations in Europe exceed the permitted levels of nitrates (50 mg NO3- L-1) or pesticides (0.1 µg L-1), respectively. Although slow sand filtration can remove nitrates via denitrification when oxygen is limited, it requires an organic carbon source. The present study evaluates the performance of the use of wood pellets and granulated cork as carbon sources in bench-scale biofilters operated under water-saturated and water-unsaturated conditions for more than 400 days. The biofilters were monitored for nitrate (200 mg L-1) and pesticide (mecoprop, diuron, atrazine, and bromacil, each at a concentration of 5 µg L-1) attenuation, as well as for the formation of nitrite and pesticide transformation products. Microbiological characterization of each biofilter was also performed. The water-saturated wood biofilter achieved the best nitrate removal (>99%), while the cork biofilters lost all denitrification power over time (from 38% to no removal). The unsaturated biofilter columns were not effective for removing nitrates (20-30% removal). As for pesticides, all the biofilters achieved high removal rates of mecoprop and diuron (>99% and >75%, respectively). Atrazine removal was better in the wood-pellet biofilters than the cork ones (68-96% vs. 31-38%). Bromacil was only removed in the water-unsaturated cork biofilter (67%). However, a bromacil transformation product was formed there. The water-saturated wood biofilter contained the highest number of denitrifying microorganisms, with Methyloversatilis as the characteristic genus. Microbial composition could explain the high removal of pesticides and nitrates achieved in the wood-pellet biofilter. Overall, the results indicate that wood-pellet biofilters operated under water-saturated conditions are a good solution for treating groundwater contaminated with nitrates and pesticides.

Impact of the antibiotic doxycycline on the D. magna reproduction, associated microbiome and antibiotic resistance genes in treated wastewater conditions.

Wastewater Treatment Plant (WWTP) effluents are important sources of antibiotics, antibiotic resistance genes (ARGs) and resistant bacteria that threaten aquatic biota and human heath. Antibiotic effects on host-associated microbiomes, spread of ARGs and the consequences for host health are still poorly described. This study investigated changes of the Daphnia magna associated microbiome exposed to the recalcitrant antibiotic doxycycline under artificial reconstituted lab water media (lab water) and treated wastewater media. D. magna individual juveniles were exposed for 10 days to treated wastewater with and without doxycycline, and similarly in lab water. We analysed 16 S rRNA gene sequences to assess changes in community structure, monitored Daphnia offspring production and quantified ARGs abundances by qPCR from both Daphnia and water (before and after the exposure). Results showed that doxycycline and media (lab water or wastewater) had a significant effect modulating Daphnia-associated microbiome composition and one of the most discriminant taxa was Enterococcus spp. Moreover, in lab water, doxycycline reduced the presence of Limnohabitans sp., which are dominant bacteria of the D. magna-associated microbiome and impaired Daphnia reproduction. Contrarily, treated wastewater increased diversity and richness of Daphnia-associated microbiome and promoted fecundity. In addition, the detected ARG genes in both lab water and treated wastewater medium included the qnrS1, sul1, and blaTEM, and the integron-related intI1 gene. The treated wastewater contained about 10 times more ARGs than lab water alone. Furthermore, there was an increase of sul1 in Daphnia cultured in treated wastewater compared to lab water. In addition, there were signs of a higher biodegradation of doxycycline by microbiomes of treated wastewater in comparison to lab water. Thus, results suggest that Daphnia-associated microbiomes are influenced by their environment, and that bacterial communities present in treated wastewater are better suited to cope with the effects of antibiotics.

The potential of wastewater grown microalgae for agricultural purposes: Contaminants of emerging concern, heavy metals and pathogens assessment.

In the coming years, the use of microalgal biomass as agricultural biofertilizers has shown promising results. The use of wastewater as culture medium has resulted in the reduction of production costs, making microalgae-based fertilizers highly attractive for farmers. However, the occurrence of specific pollutants in wastewater, like pathogens, heavy metals and contaminants of emerging concern (CECs), such as pharmaceuticals and personal care products may pose a risk on human health. This study presents an holistic assessment of the production and use of microalgal biomass grown in municipal wastewater as biofertilizer in agriculture. Results showed that pathogens and heavy metals concentrations in the microalgal biomass were below the threshold established by the European regulation for fertilizing products, except for cadmium. Regarding CECs, 25 out of 29 compounds were found in wastewater. However, only three of them (hydrocinnamic acid, caffeine, and bisphenol A) were found in the microalgae biomass used as biofertilizer. Agronomic tests were performed for lettuce growth in greenhouse. Four treatments were studied, comparing the use of microalgae biofertilizer with a conventional mineral fertilizer, and also a combination of both of them. Results suggested that microalgae can help reducing the mineral nitrogen dose, since similar fresh shoot weights were obtained in the plants grown with the different assessed fertilizers. Lettuce samples revealed the presence of cadmium and CECs in all the treatments including both negative and positive controls, which suggests that their presence was not linked to the microalgae biomass. On the whole, this study revealed that wastewater grown microalgae can be used for agricultural purposes reducing mineral N need and guaranteeing health safety of the crops.

Linking plant-root exudate changes to micropollutant exposure in aquatic plants (Lemna minor and Salvinia natans). A prospective metabolomic study.

Recent findings indicate that plant-root exudates can stimulate plant-associated microorganisms to enhance the biodegradation of contaminants in constructed wetlands. To understand this process, we studied the root-exudation changes of two aquatic plants (Lemna minor and Salvinia natans) upon micropollutants exposure (10, 100 and 1000 µg/L mixes containing naproxen, diclofenac, carbamazepine, and benzotriazole). After a 2-day exposure, plant exudates were collected, extracted and non-target analysis was performed with a gas chromatography-high resolution Orbitrap mass-spectrometer. Plants didn't show morphological or growth differences between the control and spiked reactors, but exudation changes were observed in both plants at all concentration levels. Partial least squares discriminant analysis showed that, for Lemna minor, the increase of micropollutants exposure was linked to the reduction of sugar and fatty acid exudation. This may trigger changes in the microbial community living on complex carbon forms. Instead, in Salvinia natans, micropollutants exposure was linked to the release of long-chain compounds such as cuticular waxes and sesquiterpenoids, which might be related to stress signaling. These results demonstrate that plant micropollutant-exposure at environmentally relevant concentration levels triggers changes in root exudates. This may help to design new strategies to enhance micropollutants degradation in nature based solutions such as in constructed wetlands.

Use of cover crops in vineyards to prevent groundwater pollution by copper and organic fungicides. Soil column studies.

Several fungicides, such as copper and organic products (synthetic or natural), are currently being used in vineyards to control downy mildew (Plasmopara viticola) resulting in soil, surface water, and groundwater pollution. This study aims to assess the effectiveness of using cover crops as an agricultural practice in vineyards to protect soil and groundwater pollution. For that purpose, we performed different soil column studies to quantify soil leaching of selected fungicides (copper, dimethomorph, oxathiapiprolin, zoxamide, acibenzolar-s-methyl, and laminarin) following a rainfall event after a conventional fungicide vineyard application. Two types of vineyard soils (loam and sandy-loam soil textures) and three ground covers (bare ground, monoculture cover, and polyculture cover) were assessed. These studies were completed with hydroponic assays to check the effectiveness of cover roots in the fungicide degradation. Mass balance results show that whereas 3 fungicides (Cu, zoxamide, and dimethomorph) were leached through sandy soil columns, only copper was leached from loam soil columns. The effect of cover crops was only significant for Cu and zoxamide when fungicides were applied 24 h before the rain event, reducing the fungicide leaching by 30%. Hydroponic studies showed that cover roots enhanced the kinetic rates of almost all tested fungicides by 5-467%, suggesting that they are relevant to improving the degradation of fungicides in the soil column. These results are relevant to drawing up recommendations on the use of cover crops to protect soil and groundwater pollution by fungicides.