Impact of organic soil amendments in antibiotic levels, antibiotic resistance gene loads, and microbiome composition in corn fields and crops.

The potential spreading of antibiotic resistance genes (ARG) into agricultural fields and crops represent a fundamental limitation on the use of organic fertilization in food production systems. We present here a study of the effect of spreading four types of organic soil amendments (raw pig slurry, liquid and solid fractions, and a digested derivative) on demonstrative plots in two consecutive productive cycles of corn harvest (Zea mays), using a mineral fertilizer as a control, following the application of organic amendments at 32-62 T per ha (150 kg total N/ha) and allowing 5-8 months between fertilization and harvest. A combination of qPCR and high-throughput 16S rDNA sequencing methods showed a small, but significant impact of the fertilizers in both ARG loads and microbiomes in soil samples, particularly after the second harvesting cycle. The slurry solid fraction showed the largest impact on both ARG loads and microbiome variation, whereas its digestion derivatives showed a much smaller impact. Soil samples with the highest ARG loads also presented increased levels of tetracyclines, indicating a potential dual hazard by ARG and antibiotic residues linked to some organic amendments. Unlike soils, no accumulation of ARG or antibiotics was observed in corn leaves (used as fodder) or grains, and no grain sample reached detection limits for neither parameter. These results support the use of organic soil amendments in corn crops, while proposing the reduction of the loads of ARGs and antibiotics from the fertilizers to greatly reduce their potential risk.

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.

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.

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.

Determination of the ß-glycosylate fraction of contaminants of emerging concern in lettuce (Lactuca sativa L.) grown under controlled conditions.

The uptake of a large variety of contaminants of emerging concern (CECs) by crops has already been reported, and the occurrence of phase II metabolites or conjugates has only been detected in plant cell cultures. However, the extent of their formation under cropping conditions is largely unknown. In this study, an analytical strategy to assess the conjugation of 11 CECs in lettuce (Lactuca sativa L.) irrigated with different concentrations (0, 0.05, 0.5, 5, and 50 µg L-1) of CECs was developed. The methodology involved enzymatic digestion with ß-glucosidase to obtain the total fraction (free form + conjugates) of CECs. The conjugation fraction was then obtained based on the difference. The highest extent of conjugation (i.e., 27 to 83%) was found with the most hydrophobic compounds, such as bisphenol A, carbamazepine, methyl paraben, and triclosan. So, the CEC conjugate fraction cannot be neglected in the estimate of human daily intake.

Distribution and Sources of Petroleum Hydrocarbons in Recent Sediments of the Imo River, SE Nigeria.

The distribution of aliphatic and aromatic hydrocarbons in surface sediments of the lower course of the Imo River (Nigeria) was investigated to determine the sources and fate of these compounds. The aliphatic fraction is characterized by a widespread contribution of highly weathered/biodegraded hydrocarbon residues (reflected in the absence of prominent n-alkane peaks coupled with the presence of 17α(H),21ß(H)-25-norhopane, an indicator of heavy hydrocarbon biodegradation) of Nigerian crude oils (confirmed by the occurrence of 18α(H)-oleanane, a compound characteristic of oils of deltaic origin). The concentrations of polycyclic aromatic hydrocarbons (PAHs) ranging from 48 to 117 ng/g dry weight (dw; ∑13PAHs) indicate a moderate pollution, possibly lowered by the sandy lithology and low organic carbon (OC) content of the sediments. Concentrations slightly decrease towards the estuary of the river, probably due to the fact that these stations are affected by tidal flushing of pollutants adsorbed on sediment particles and carried away by occasional storm to the Atlantic Ocean. A number of PAH ratios, including parent/alkylated and isomeric compounds, indicates a predominance of petrogenic sources, with a low contribution of pyrolytic inputs, particularly of fossil fuel combustion. On the basis of OC/ON (>10) and Per/ΣPAHpenta- (>10) values, a diagenetic terrigenous OC was proposed as a source of perylene to the river.

Chemometrics-assisted effect-directed analysis of crude and refined oil using comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry.

An effect-directed analysis (EDA) of fresh and artificially weathered (evaporated, photooxidized) samples of North Sea crude oil and residual heavy fuel oil is presented. Aliphatic, aromatic, and polar oil fractions were tested for the presence of aryl hydrocarbon receptor (AhR) agonist and androgen receptor (AR) antagonist, demonstrating for the first time the AR antagonist effects in the aromatic and, to a lesser extent, polar fractions. An extension of the typical EDA strategy to include an N-way partial least-squares (N-PLS) model capable of relating the comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC × GC-TOFMS) data set to the bioassay data obtained from normal-phase LC fractions is proposed. The predicted AhR binding effects in the fresh and artificially weathered aromatic oil fractions facilitated the identification of alkyl-substituted three- and four-ring aromatic systems in the active fractions through the weighting of their contributions to the observed effects. A N-PLS chemometric model is demonstrated as a potentially useful strategy for future EDA studies that can streamline the compound identification process and provide additional reduction of samples' complexity. The AhR binding effects of the suspected compounds predicted by N-PLS and identified by GC × GC-TOFMS were confirmed using quantitative structure-activity relationship (QSAR) estimates.

Input and leaching potential of copper, zinc, and selenium in agricultural soil from swine slurry.

Trace elements, such as copper, zinc, and selenium, used as feed additives were determined in samples of both fresh (N = 14) and anaerobically digested (N = 6) swine slurry collected on medium- to large-size farms in northeast Spain. Considering both fresh and anaerobically digested samples, mean concentrations of zinc (1,500 mg kg(-1) dry mass [dm]) were greater than those of copper (mean 239 mg kg(-1 )dm), and the selenium concentrations detected were even lower (mean 139 µg kg(-1) dm). Zinc concentrations were significantly greater in anaerobically digested samples, whereas no significant differences were found for copper or selenium. In addition, the leaching potential of zinc, copper, and selenium in cropped (lettuce heart) and uncropped experimental units subject to drip irrigation was assessed in a greenhouse experiment. Generally, the addition of swine slurry to soil (1.7 g kg(-1) dm) significantly increased zinc, copper, and selenium concentrations in leachates, which decreased in accordance with the volume of leachate eluted. Under the experimental conditions, the leaching potential of zinc and selenium was more strongly correlated with bulk parameters directly associated with the composition of the pig slurry (dissolved organic carbon, electrical conductivity, and ammonium), whereas copper mobility was more strongly associated with the crop root exudates. Although selenium has been shown to be mobile in soil, the selenium content found in the leachates did not pose any appreciable risk according to current drinking water standards.

Assessment of a dielectric barrier discharge plasma reactor at atmospheric pressure for the removal of bisphenol A and tributyltin.

The ability of a laboratory-scale dielectric barrier discharge (DBD) nonthermal plasma reactor at atmospheric pressure was assessed for the removal of bisphenol A (1 mg L(-1)) and tributyltin (10 mg L(-1)) from aqueous solutions. The elimination of both the compounds followed an exponential decay equation, and a first-order degradation kinetics was proposed for them (k = 0.662 min(-1) for bisphenol A and k = 0.800 min(-1) for tributyltin), reaching in both cases about 96% removal after 5-min treatment. Accordingly, plasma DBD reactors could be used as alternative advanced oxidation technologies for the removal of some persistent and toxic pollutants from water and wastewater, although further research should be performed to evaluate the effluent toxicity.

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.

Towards universal wavelength-specific photodegradation rate constants for methyl mercury in humic waters, exemplified by a Boreal lake-wetland gradient.

We report experimentally determined first-order rate constants of MeHg photolysis in three waters along a Boreal lake-wetland gradient covering a range of pH (3.8-6.6), concentrations of total organic carbon (TOC 17.5-81 mg L(-1)), total Fe (0.8-2.1 mg L(-1)), specific UV254 nm absorption (3.3-4.2 L mg(-1) m(-1)) and TOC/TON ratios (24-67 g g(-1)). Rate constants determined as a function of incident sunlight (measured as cumulative photon flux of photosynthetically active radiation, PAR) decreased in the order dystrophic lake > dystrophic lake/wetland > riparian wetland. After correction for light attenuation by dissolved natural organic matter (DOM), wavelength-specific (PAR: 400-700 nm, UVA: 320-400 nm and UVB: 280-320 nm) first-order photodegradation rate constants (kpd) determined at the three sites were indistinguishable, with average values (± SE) of 0.0023 ± 0.0002, 0.10 ± 0.024 and 7.2 ± 1.3 m(2) E(-1) for kpdPAR, kpdUVA, and kpdUVB, respectively. The relative ratio of kpdPAR, kpdUVA, and kpdUVB was 1:43:3100. Experiments conducted at varying MeHg/TOC ratios confirm previous suggestions that complex formation with organic thiol groups enhances the rate of MeHg photodegradation, as compared to when O and N functional groups are involved in the speciation of MeHg. We suggest that if the photon fluxes of PAR, UVA, and UVB radiation are separately determined and the wavelength-specific light attenuation is corrected for, the first-order rate constants kpdPAR, kpdUVA, and kpdUVB will be universal to waters in which DOM (possibly in concert with Fe) controls the formation of ROS, and the chemical speciation of MeHg is controlled by the complexation with DOM associated thiols.

Solving chromatographic challenges in comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry using multivariate curve resolution-alternating least squares.

Multivariate curve resolution-alternating least squares (MCR-ALS) analysis is proposed to solve chromatographic challenges during two-dimensional gas chromatography-time-of-flight mass spectrometry (GC × GC-TOFMS) analysis of complex samples, such as crude oil extract. In view of the fact that the MCR-ALS method is based on the fulfillment of the bilinear model assumption, three-way and four-way GC × GC-TOFMS data are preferably arranged in a column-wise superaugmented data matrix in which mass-to-charge ratios (m/z) are in its columns and the elution times in the second and first chromatographic columns are in its rows. Since m/z values are common for all measured spectra in all second-column modulations, unavoidable chromatographic challenges such as retention time shifts within and between GC × GC-TOFMS experiments are properly handled. In addition, baseline/background contributions can be modeled by adding extra components to the MCR-ALS model. Another outstanding aspect of MCR-ALS analysis is its extreme flexibility to consider all samples (standards, unknowns, and replicates) in a single superaugmented data matrix, allowing joint analysis. In this way, resolution, identification, and quantification results can be simultaneously obtained in a very fast and reliable way. The potential of MCR-ALS analysis is demonstrated in GC × GC-TOFMS analysis of a North Sea crude oil extract sample with relative errors in estimated concentrations of target compounds below 6.0 % and relative standard deviations lower than 7.0 %. The results obtained, along with reasonable values for the lack of fit of the MCR-ALS model and high values of the reversed match factor in mass spectra similarity searches, confirm the reliability of the proposed strategy for GC × GC-TOFMS data analysis.

Compositional properties characterizing commonly transported oils and controlling their fate in the marine environment.

Oil spills relating to shipping incidents remain of substantial concern with respect to marine pollution. Whilst most frequently a reactive approach is adopted in post-incident monitoring (for the specific product involved), this paper reports important physical and compositional characteristics of commonly transported oils and oil products to afford pro-active assessments. These properties include specific gravity, viscosity, elemental composition and, of particular relevance, the relative class compositions between aliphatics, aromatics, resins and asphaltenes. The latter were determined experimentally using thin layer chromatography with flame ionization detection. Diagnostic ratios of specific compounds are reported, statistically analysed, and their significance in identification of different oil types and the weathering processes is discussed. The influence of the properties on fates under different environmental conditions (selected to represent contrasting European regional seas) are examined using the NOAA Automated Data Inquiry for Oil Spills (ADIOS2) model. Relative contributions of the different environmental conditions and properties to the fate of the oil at sea are discussed.

Post-incident monitoring to evaluate environmental damage from shipping incidents: chemical and biological assessments.

Oil and chemical spills in the marine environment are an issue of growing concern. Oil exploration and exploitation is moving from the continental shelf to deeper waters, and to northern latitudes where the risk of an oil spill is potentially greater and may affect pristine ecosystems. Moreover, a growing number of chemical products are transported by sea and maritime incidents of hazardous and noxious substances (HNS) are expected to increase. Consequently, it seems timely to review all of the experience gained from past spills to be able to cope with appropriate response and mitigation strategies to combat future incidents. Accordingly, this overview is focused on the dissemination of the most successful approaches to both detect and assess accidental releases using chemical as well as biological approaches for spills of either oil or HNS in the marine environment. Aerial surveillance, sampling techniques for water, suspended particles, sediments and biota are reviewed. Early warning bioassays and biomarkers to assess spills are also presented. Finally, research needs and gaps in knowledge are discussed.

Assessment of a novel microalgae-cork based technology for removing antibiotics, pesticides and nitrates from groundwater.

Groundwater pollution has increased in recent years due to the intensification of agricultural and livestock activities. This results in a significant reduction in available freshwater resources. Here, we have studied the long term assessment of a green technology (1-4 L/day) based on a photobioreactor (PBR) containing immobilised microalgae-bacteria in polyurethane foam (PF) followed by a cork filter (CF) for removing nitrates, pesticides (atrazine and bromacil), and antibiotics (sulfamethoxazole and sulfacetamide) from groundwater. The prototype was moderately effective for removing nitrates (58%) at an HRT of 8 days, while its efficiency decreased at a HRT of 4 and 2 days (<20% removal). The combined use of PBR-CF enabled antibiotics and pesticides to be attenuated by up to 95% at an HRT of 8 days, but their attenuation decreased with shorter HRT, with pesticides being the compounds most affected (reducing from 97 to 98% at an HRT of 8 days to 23-45% at an HRT of 2 days). Pesticide transformation products were identified after the CF, supporting biodegradation as the main attenuation process. A gene-based metataxonomic assessment linked the attenuation of micropollutants to the presence of specific pesticide biodegradation species (e.g. genus Phenylobacterium, Sphingomonadaceae, and Caulobacteraceae). Therefore, the results highlighted the potential use of microalgae and cork to treat polluted groundwater.

Polyphasic approach for assessing changes in an autochthonous marine bacterial community in the presence of Prestige fuel oil and its biodegradation potential.

A laboratory experiment was conducted to identify key hydrocarbon degraders from a marine oil spill sample (Prestige fuel oil), to ascertain their role in the degradation of different hydrocarbons, and to assess their biodegradation potential for this complex heavy oil. After a 17-month enrichment in weathered fuel, the bacterial community, initially consisting mainly of Methylophaga species, underwent a major selective pressure in favor of obligate hydrocarbonoclastic microorganisms, such as Alcanivorax and Marinobacter spp. and other hydrocarbon-degrading taxa (Thalassospira and Alcaligenes), and showed strong biodegradation potential. This ranged from >99% for all low- and medium-molecular-weight alkanes (C(15)-C(27)) and polycyclic aromatic hydrocarbons (C(0)- to C(2)- naphthalene, anthracene, phenanthrene, dibenzothiophene, and carbazole), to 75-98% for higher molecular-weight alkanes (C(28)-C(40)) and to 55-80% for the C(3) derivatives of tricyclic and tetracyclic polycyclic aromatic hydrocarbons (PAHs) (e.g., C(3)-chrysenes), in 60 days. The numbers of total heterotrophs and of n-alkane-, aliphatic-, and PAH degraders, as well as the structures of these populations, were monitored throughout the biodegradation process. The salinity of the counting medium affects the counts of PAH degraders, while the carbon source (n-hexadecane vs. a mixture of aliphatic hydrocarbons) is a key factor when counting aliphatic degraders. These limitations notwithstanding, some bacterial genera associated with hydrocarbon degradation (mainly belonging to α- and γ-Proteobacteria, including the hydrocarbonoclastic Alcanivorax and Marinobacter) were identified. We conclude that Thalassospira and Roseobacter contribute to the degradation of aliphatic hydrocarbons, whereas Mesorhizobium and Muricauda participate in the degradation of PAHs.

Local and downstream cumulative effects of traditional meadow management on stream-water quality and multiple riparian taxa.

Water quality and riparian communities are among the most affected stream components by agriculture. However, little is known about the effects of riparian management for both aquatic and terrestrial taxa at different spatial scales. Here, we surveyed aquatic (diatoms) and terrestrial taxa (bryophytes, vascular plants, litter-dwelling snails, and ground and volant arthropods), to compare the abundance and richness of riparian taxa and chemical quality between reference and exposed sites in two stream reaches each of c. 3.5 km in northwestern Spain. Impacts in exposed sites were mainly due to traditional farming practices (TFPs), which included traditional meadow management, weirs built for now-unused water mills and sporadic timber harvesting. Therefore, we measured ten covariates and predictors related to the intensification of TFPs at local and within-stream scales and explored associations with riparian and water-quality measures to study the potential effects of TFPs in more detail. Reference and exposed sites did not differ significantly in water properties (diatom-biotic indices, conductivity, total organic carbon and nitrates), but exposed sites had less concentrations of soil metals Cd, Cu, Ni and Zn and less cover and richness of riparian trees, as inferred by the index QBR. Exposed sites had more volant insect decomposers and reference sites a greater abundance or richness of snails, ground predators and decomposers. Bryophyte richness was greater in reference sites. Our inferences may inform the joint cumulative downstream effects of weirs, meadow uses and riparian alterations but were generally consistent with most riparian taxa benefiting from having larger forested areas. Given the contrasting responses among taxa, we argue that land snails, terrestrial flies, and centipedes may be valuable additions to current riparian assessments mostly based on plants, beetles and spiders as indicator taxa. Our study also suggests caution when inferring farming impacts on streams from the surface area of pastoral land.

Part-per-trillion determination of pharmaceuticals, pesticides, and related organic contaminants in river water by solid-phase extraction followed by comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry.

An analytical procedure based on comprehensive two-dimensional gas chromatography (GC x GC) coupled with time-of-flight mass spectrometry (TOF-MS) for the simultaneous determination of 97 organic contaminants at trace concentration in river water is presented. The target analytes included 13 pharmaceuticals, 18 plasticizers, 8 personal care products, 9 acid herbicides, 8 triazines, 10 organophosphorous compounds, 5 phenylureas, 12 organochlorine biocides, 9 polycyclic aromatic hydrocarbons (PAHs), and 5 benzothiazoles and benzotriazoles. The best resolution of the target analytes in the contour plots was obtained when a nonpolar stationary phase was used in the first dimension and polar one in the second. However, in the opposite configuration, polar-nonpolar, the retention time in the second dimension exhibited a strong correlation with the log Kow (p < 0.01), and it was proposed as an additional identification criteria. The developed methodology is based on a polymeric solid-phase extraction followed by in GC-port methylation and GC x GC/TOF-MS determination. Moreover, limits of detection (LODs) and quantification (LOQs) ranged from 0.5 to 100 ng/L and from 2 to 185 ng/L, respectively. Repeatability was always lower than 20%. Finally, the developed method has been successfully applied to the determination of incurred target analytes in four river waters subjected to a different anthropogenic pressure.