Metabolomic and phenotypic implications of the application of fertilization products containing microcontaminants in lettuce (Lactuca sativa)

Cultivation practice using organic amendments is plausible to ensure global food security. However, plant abiotic stress due to the presence of metals and organic microcontaminants (OMCs) in fertilization products cannot be overlooked. In this study, we monitored lettuce metabolism and phenotypic response following the application of either sewage sludge (SS), the organic fraction of municipal solid waste, swine manure (SM), chemical fertilizers (CF), or no amendment (C) in a greenhouse facility.Postprint (published version

Occurrence and bioaccumulation of chemical contaminants in lettuce grown in peri-urban horticulture

Peri-urban horticulture performs environmental and socio-economic functions and provides ecological services to nearby urban areas. Nevertheless, industrialization and water pollution have led to an increase in the exposure of peri-urban vegetables to contaminants such as trace elements (TEs) and organic microcontaminants (OMCs). In this study, the occurrence of chemical contaminants (i.e., 16 TEs, 33 OMCs) in soil and lettuce leaves from 4 farm fields in the peri-urban area of the city of Barcelona was assessed. A rural site, outside the peri-urban area of influence, was selected for comparison. The concentration of TEs and OMCs ranged from non-detectable to 803¿mg/kg¿dw and from non-detectable to 397¿µg/kg¿dw respectively in the peri-urban soil, and from 6¿·¿10-5 to 4.91¿mg/kg¿fw and from non-detectable to 193¿µg/kg¿fw respectively in lettuce leaves. Although the concentration of Mo, Ni, Pb, and As in the soil of the peri-urban area exceeded the environmental quality guidelines, their occurrence in lettuce complied with human food standards (except for Pb). The many fungicides (carbendazim, dimetomorph, and methylparaben) and chemicals released by plastic pipelines (tris(1-chloro-2-propyl)phosphate, bisphenol F, and 2-mercaptobenzothiazole) used in agriculture were prevalent in the soil and the edible parts of the lettuce. The occurrence of these chemical pollutants in the peri-urban area did not affect the chlorophyll, lipid, or carbohydrate content of the lettuce leaves. PCA (Principal Component Analysis) showed that soil pollution, fungicide application, and irrigation water quality are the most relevant factors determining the presence of contaminants in crops.Postprint (updated version

Effects of tetracycline, sulfonamide, fluoroquinolone, and lincosamide load in pig slurry on lettuce: Agricultural and human health implications

The application of pig slurry as fertilizer in agriculture provides nutrients, but it can also contain veterinary medicines, including antibiotic residues (ABs), which can have an ecotoxicological impact on agroecosystems. Furthermore, uptake, translocation, and accumulation of ABs in crops can mobilize them throughout the food chain. This greenhouse study aims to assess AB uptake from soil fertilized with pig slurry and its phenotypical effects on Lactuca sativa LThe authors gratefully acknowledge the financial support of the Spanish Ministry of Science and Innovation through project AGL 2017- 89518-R. IDAEA-CSIC is a Severo Ochoa Centre of Excellence (Spanish Ministry of Science and Innovation, Project CEX 2018-000794-S). Monica Escola Casas wishes to thank the Beatriu de Pinos 2018 grant program (MSCA grant agreement number 801370) for the funding. The authors likewise thank Miquel Massip, Daniel Fenero, and Nerea Gran- ados for their technical assistance in the greenhouse facilityPostprint (published version

Implications of the use of organic fertilizers for antibiotic resistance gene distribution in agricultural soils and fresh food products. A plot-scale study

The spread of antibiotic resistance genes (ARG) into agricultural soils, products, and foods severely limits the use of organic fertilizers in agriculture. In order to help designing agricultural practices that minimize the spread of ARG, we fertilized, sown, and harvested lettuces and radish plants in experimental land plots for two consecutive agricultural cycles using four types of fertilizers: mineral fertilization, sewage sludge, pig slurry, or composted organic fraction of municipal solid waste. The analysis of the relative abundances of more than 200,000 ASV (Amplicon Sequence Variants) identified a small, but significant overlap (<10%) between soil's and fertilizer microbiomes. Clinically relevant ARG were found in higher loads (up to 100 fold) in fertilized soils than in the initial soil, particularly in those treated with organic fertilizers, and their loads grossly correlated to the amount of antibiotic residues found in the corresponding fertilizer. Similarly, low, but measurable ARG loads were found in lettuce (tetM, sul1) and radish (sul1), corresponding the lowest values to samples collected from minerally fertilized fields. Comparison of soil samples collected along the total period of the experiment indicated a relatively year-round stability of soil microbiomes in amended soils, whereas ARG loads appeared as unstable and transient. The results indicate that ARG loads in soils and foodstuffs were likely linked to the contribution of bacteria from organic fertilizer to the soil microbiomes, suggesting that an adequate waste management and good pharmacological and veterinarian practicesmay significantly reduce the presence of these ARGs in agricultural soils and plant products.Postprint (published version

Occurrence of antibiotics in Lettuce (Lactuca sativa L.) and Radish (Raphanus sativus L.) following organic soil fertilisation under plot-scale conditions: Crop and human health implications

Recent studies have demonstrated the crop uptake of antibiotics (ABs) from soils treated with AB-carrying fertilisers. However, there is a lack of plot-scale studies linking their effects at the agronomic and metabolomic/transcriptomic level to their impact on human health. This paper assesses the plant uptake of 23 ABs following two productive cycles of lettuce and radish cropped with sewage sludge, pig slurry, the organic fraction of municipal solid waste, or chemical fertilisation under plot-scale conditions (32 plots spanning 3-10 m2 each). AB uptake by plants depended on both the vegetable and the AB class and was higher in radish than in lettuce edible parts. Levels ranged from undetectable to up to 76 ng/g (fresh weight). Repetitive organic fertilisation resulted in an increase in the concentration of ABs in lettuce leaves, but not in radish roots. Significant metabolomic and transcriptomic changes were observed following soil fertilisation. Nevertheless, a human health risk assessment indicates that the occurrence of ABs in lettuce or radish edible parts does not pose any risk. To our knowledge, this is the first holistic plot-scale study demonstrating that the use of organic fertilisers containing ABs is safe for crop security and human health.The authors gratefully acknowledge the financial support of the Spanish Ministry of Science and Innovation through projects AGL2017- 89518-R and RTI2018-096175-B-I00. IDAEA-CSIC is a Severo Ochoa Centre of Excellence (Spanish Ministry of Science and Innovation, Project CEX2018-000794-S). Mònica Escolà Casas wishes to thank the Beatriu de Pinós 2018 grant programme (MSCA grant agreement number 801370) for the funding.Peer ReviewedPostprint (published version

Co-digestion of microalgae and primary sludge: effect on biogas production and microcontaminants removal

Microalgal-based wastewater treatment plants are conceived as low cost and low energy consuming systems. The operation of these plants involves the management of primary sludge and microalgal biomass. The aim of this study is to analyse the anaerobic co-digestion of both by-products in terms of biogas production and contaminants of emerging concern removal. The co-digestion of microalgae and primary sludge (25/75% on a volatile solids basis) was investigated in continuous reactors and compared to microalgae mono-digestion at a hydraulic retention time of 20 days. Results showed how the co-digestion enhanced the anaerobic digestion of microalgal biomass, since primary sludge is a more readily biodegradable substrate, which increased the methane production by 65% and reduced the risk of ammonia toxicity. Regarding the contaminants, musk fragrances (galaxolide and tonalide) and triclosan showed the highest abundance on primary sludge (0.5–25 µg/g TS), whereas caffeine, methyl dihydrojasmonate and triphenyl phosphate were barely detected in both substrates (<0.1 µg/g TS). The removal of these contaminants was compound-depending and ranged from no removal to up to 90%. On the whole, microalgae mono-digestion resulted in a higher removal of selected contaminants than the co-digestion with primary sludge.Peer Reviewe

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.This research was funded by European Union's Horizon 2020 Research and Innovation Program within the framework of the INCOVER project (GA 689242); by the Spanish Ministry of Science, Innovation and Universities, the Research National Agency (AEI), and the European Regional Development Fund (FEDER) within the projects AL4BIO (RTI2018-099495-B-C21) and SIMAGUA (CTM2017-91355-EXP), and by the Government of Catalonia (Consolidated Research Group 2017 SGR 1029). M.J. García-Galán, E. Uggetti and R. Díez-Montero would like to thank the Spanish Ministry of Economy and Competitiveness for their research grants (IJCI-2017-34601, RYC2018-025514-I and FJCI-2016-30997, respectively).This research was funded by European Union's Horizon 2020 Research and Innovation Program within the framework of the INCOVER project (GA 689242); by the Spanish Ministry of Science, Innovation and Universities, the Research National Agency (AEI), and the European Regional Development Fund (FEDER) within the projects AL4BIO (RTI2018-099495-B-C21) and SIMAGUA (CTM2017-91355-EXP), and by the Government of Catalonia (Consolidated Research Group 2017 SGR 1029). M.J. García-Galán, E. Uggetti and R. Díez-Montero would like to thank the Spanish Ministry of Economy and Competitiveness for their research grants (IJCI-2017-34601, RYC2018-025514-I and FJCI-2016-30997, respectively).Peer ReviewedPostprint (author's final draft

Codigestión de microalgas y fango primario: producción de energía y reutilización del digestato

Los sistemas de tratamiento de aguas residuales con microalgas combinan el tratamiento de aguas residuales y la producción de bioenergía. Para este fin, una de las tecnologías más apropiadas es la digestión anaerobia, obteniendo como productos el biogás y un digestato estabilizado y rico en nutrientes que puede ser reutilizado en la agricultura. La operación de estas plantas requiere tanto la gestión del lodo primario como de la biomasa microalgal (lodo secundario), por lo que la codigestión de ambos sustratos en un mismo reactor podría mejorar el rendimiento de la digestión y a la vez facilitar la gestión de estos subproductos. Por otro lado, la idoneidad de este digestato para su reutilización agrícola aún no se ha evaluado, pero su valorización puede ser de gran interés y permite cerrar ciclo de todo el proceso. El objetivo del presente estudio es investigar la codigestión anaeróbica de la biomasa microalgal y el fango primario en términos de producción de biogás y aprovechamiento del digestato.Postprint (author's final draft

Occurrence of chemical contaminants in peri-urban agricultural irrigation waters and assessment of their phytotoxicity and crop productivity

Water scarcity and water pollution have increased the pressure on water resources worldwide. This pressure is particularly important in highly populated areaswherewater demand exceeds the available natural resources. In this regard, water reuse has emerged as an excellent water source alternative for peri-urban agriculture. Nevertheless, itmust copewith the occurrence of chemical contaminants, ranging fromtrace elements (TEs) to organic microcontaminants. In this study, chemical contaminants (i.e., 15 TEs, 34 contaminants of emerging concern (CECs)), bulk parameters, and nutrients from irrigation waters and crop productivity (Lycopersicon esculentum Mill. cv. Bodar and Lactuca sativa L. cv. Batavia) were seasonally surveyed in 4 farm plots in the peri-urban area of the city of Barcelona. A pristine site, where rain-groundwater is used for irrigation, was selected for background concentrations. The average concentration levels of TEs and CECs in the irrigation water impacted by treated wastewater (TWW) were 3 (35±75 µg L-1) and 13 (553±1050 ng L-1) times higher than at the pristine site respectively. Principal component analysiswas used to classify the irrigationwaters by chemical composition. To assess the impact of the occurrence of these contaminants on agriculture, a seed germination assay (Lactuca sativa L) and real field-scale study of crop productivity (i.e., lettuce and tomato)were used. Although irrigation waters from the peri-urban area exhibited a higher frequency of detection and concentration of the assessed chemical contaminants than those of the pristine site (P1), no significant differences were found in seed phytotoxicity or crop productivity. In fact, the crops impacted by TWW showed higher productivity than the other farm plots studied, which was associated with the higher nutrient availability for plants.Peer ReviewedPostprint (updated version

Occurrence and bioaccumulation of chemical contaminants in lettuce grown in peri-urban horticulture

Peri-urban horticulture performs environmental and socio-economic functions and provides ecological services to nearby urban areas. Nevertheless, industrialization and water pollution have led to an increase in the exposure of peri-urban vegetables to contaminants such as trace elements (TEs) and organic microcontaminants (OMCs). In this study, the occurrence of chemical contaminants (i.e., 16 TEs, 33 OMCs) in soil and lettuce leaves from 4 farm fields in the peri-urban area of the city of Barcelona was assessed. A rural site, outside the peri-urban area of influence, was selected for comparison. The concentration of TEs and OMCs ranged from non-detectable to 803¿mg/kg¿dw and from non-detectable to 397¿µg/kg¿dw respectively in the peri-urban soil, and from 6¿·¿10-5 to 4.91¿mg/kg¿fw and from non-detectable to 193¿µg/kg¿fw respectively in lettuce leaves. Although the concentration of Mo, Ni, Pb, and As in the soil of the peri-urban area exceeded the environmental quality guidelines, their occurrence in lettuce complied with human food standards (except for Pb). The many fungicides (carbendazim, dimetomorph, and methylparaben) and chemicals released by plastic pipelines (tris(1-chloro-2-propyl)phosphate, bisphenol F, and 2-mercaptobenzothiazole) used in agriculture were prevalent in the soil and the edible parts of the lettuce. The occurrence of these chemical pollutants in the peri-urban area did not affect the chlorophyll, lipid, or carbohydrate content of the lettuce leaves. PCA (Principal Component Analysis) showed that soil pollution, fungicide application, and irrigation water quality are the most relevant factors determining the presence of contaminants in crops