Ecotoxicological relevance of glyphosate and flazasulfuron to soil habitat and retention functions - Single vs combined exposures.

Glyphosate (GLY) and flazasulfuron (FLA) are two non-selective herbicides commonly applied together. However, research focused on their single and combined ecotoxicological impacts towards non-target organisms is still inconclusive. Therefore, this study aimed to test their single effects on soil's habitat and retention functions, and to unravel their combined impacts to earthworms and terrestrial plants. For this, ecotoxicological assays were performed with plants (Medicago sativa), oligochaetes (Eisenia fetida) and collembola (Folsomia candida). Soil elutriates were also prepared and tested in macrophytes (Lemna minor) and microalgae (Raphidocelis subcapitata). FLA (82-413 µg kg-1) reduced earthworms' and collembola's reproduction and severely impaired M. sativa growth, being much more toxic than GLY (up to 30 mg kg-1). In fact, the latter only affected plant growth (≥ 9 mg kg-1) and earthworms (≥ 13 mg kg-1), especially at high concentrations, with no effects on collembola. Moreover, only elutriates from FLA-contaminated soils significantly impacted L. minor and R. sucapitata. The experiments revealed that the co-exposure to GLY and FLA enhanced the toxic effects of contaminated soils not only on plants but also on earthworms'. However, such increase in toxicity was dependent on GLY residual concentrations in soils. Overall, this work underpins that herbicides risk assessment should consider herbicides co-exposures, since the evaluation of single exposures is not representative of current phytosanitary practices and of the potential effects under field conditions, where residues of different compounds may persist in soils.

Chemical characterization of riverine sediments affected by wastewater treatment plant effluent discharge.

This study was aimed at assessing the contribution of wastewater treatment effluents to the contamination profile of the sediments of receiving waterways. Three wastewater treatment plants (WWTP) were addressed, encompassing different population equivalent sizes, urbanization degrees and treatment methods translating differences in expected contamination patterns. Within each WWTP system, the assessment targeted the effluent and sediment samples collected upstream and downstream the effluent discharge point; contaminants belonging to several concerning chemical classes (metals and metalloids; pesticides; pharmaceuticals and personal care products, PPCPs; and polycyclic aromatic hydrocarbons, PAHs) were quantified both in effluent and sediment samples. Clear associations between contaminants present in the effluent and corresponding sediment samples were not always verified. In fact, a noticeable difference between the number or abundance of contaminants detected in effluents and in sediments, suggesting that effluents are not always the most likely source (e.g. PAHs). However, sediment contaminants that were likely sourced by the effluents were also identified (e.g. PPCPs). Sediment analysis offers an important historical view of contamination, especially in flowing recipient ecosystems where any characterization over the water matrix is ephemeral and linking exclusively to the moment of sampling. Hence, sediments should be considered for the establishment of WWTP operational benchmarks regulating the emission of contaminants, which is currently focused mostly on effluent composition thus potentially over/underestimating the longer-term impact of effluent discharge in the recipient waterways.

Nano-Fe2O3 as a tool to restore plant growth in contaminated soils - Assessment of potentially toxic elements (bio)availability and redox homeostasis in Hordeum vulgare L.

This work aimed to evaluate the potential of Fe2O3 nanoparticles (nano-Fe2O3) to alleviate potentially toxic elements (PTEs) - induced stress in barley plants (Hordeum vulgare L.), focusing on bioaccumulation patterns and on plant growth and redox homeostasis. To achieve this goal, plants grew in two agricultural soils, contaminated by different levels of PTEs, collected from an industrial area, previously amended, or not, with 1% (w/w) nano-Fe2O3. After 14 d of growth, biometric parameters were evaluated, along with the analysis of PTEs bioaccumulation and biochemical endpoints. After exposure to contaminated soils, plant development was greatly affected, as evidenced by significant decreases in root length and biomass production. However, upon co-treatment with nano-Fe2O3, lower inhibitory effects on biometric parameters were observed. Regarding the oxidative damage, both soils led to increases in lipid peroxidation and superoxide anion concentration, though hydrogen peroxide levels were only increased in the most contaminated soil. In general, these changes in the oxidative stress markers were accompanied by an upregulation of different antioxidant mechanisms, whose efficiency was even more powerful upon soil amendment with nano-Fe2O3, thus lowering PTEs-induced oxidative damage. Altogether, the present study revealed that nano-Fe2O3 can protect the growth of barley plants under contaminated soils.

Sources of carbohydrates on bulk deposition in South-Western of Europe.

Scarce information is available concerning the presence of carbohydrates in rainwater. The existence of carbohydrates in bulk deposition at the town of Estarreja (Portugal), at industrial (I) and background (BG) locals, in winter and spring seasons 2016, was assessed. Seventeen carbohydrates and related compounds were identified: monosaccharides (ribose, arabinose, xylose, glucose, galactose, fructose), disaccharides (sucrose, trehalose, maltose, cellobiose), polyols (arabinitol, xylitol, myo-inositol, mannitol, glucitol, maltitol), and the anhydromonosaccharide levoglucosan. Higher content of carbohydrates was observed in spring (BG: 670 nM; I: 249 nM) than in winter (BG: 168 nM; I: 195 nM), and fructose was the carbohydrate with the highest contribution in both seasons (spring: 32%/44% (I/BG); winter: 24% (at both sites)). Fructose, myo-inositol, glucose and sucrose showed higher volume-weighted averages (VWA) concentrations in spring than in winter, possibly due to biogenic emissions typical of spring, such as pollen, and fungal spores for myo-inositol. Fructose may have derived from isomerization of glucose in biomass burning, namely in winter. Levoglucosan and galactose presented higher VWA concentration in winter than in spring, suggesting a seasonal effect related with the biomass combustion. The carbohydrates VWA concentrations were similar for samples associated with maritime and terrestrial air masses, indicating that local sources were their main contributors. Source assessment of carbohydrates by factor analysis suggested: biogenic sources for the arabinitol, myo-inositol, glucose, fructose and sucrose; soil dust for the trehalose; and anthropogenic sources from biomass burning for the galactose, arabinose and levoglucosan. The bulk deposition showed to be fundamental on removing carbohydrates from the atmosphere.

Spatial distribution of organic and inorganic contaminants in Ria de Aveiro Lagoon: A fundamental baseline dataset.

This paper presents baseline data of sediments characterization from Ria de Aveiro lagoon (Portugal). Twenty-two intertidal surface sediments were collected and analysed for their pseudo-total content of 25 major and trace elements and for their total content of PAHs (Sum16) and PCBs (Sum13). The fine fraction percentage and total organic carbon content were also determined. Ria de Aveiro Lagoon has been the target of several studies during the last decades, and it has been pointed out as a hot spot in terms of contamination García-Seoane et al., 2016. Nevertheless, data about sediments contamination is mostly focused on Hg Lillebø et al.,2011, and in a specific area of the lagoon Grilo et al., 2013, and there is not a clear and global picture about the spatial distribution of other elements, and especially of organic contaminants. Thus, this baseline dataset is of utmost importance for researchers and planners and it can be used to monitor the effect of disturbances, such as: dredging activities; hydrodynamic changes (either due to human activities or extreme events); external inputs (new discharges from industries and wastewater plants, etc).

Source and pathway analysis of lead and polycyclic aromatic hydrocarbons in Lisbon urban soils.

One hundred soil samples were collected from urban spaces, in Lisbon, Portugal, in two surveys that were carried out in consecutive years, to assess the potential adverse human health effects following exposure to potentially toxic elements and organic compounds in the urban soils. The study hereby described follows on from the earlier work of the authors and aims at performing a source-pathway-fate analysis of lead (Pb) and polycyclic aromatic hydrocarbons (PAHs) in the urban soils in order to increase current knowledge on factors influencing exposure of the population. Various techniques were combined to achieve the proposed goal. Geogenic and anthropogenic sources were apportioned by means of Pb isotope mixing models. Isotope data was further coupled with geographic information system mapping to assess local mixed sources of Pb and PAHs. Unleaded vehicle exhaust and cement production show the largest relative contribution to the total soil-Pb, but their respective importance depends on factors such as location and urban landscape. The primary sources of PAHs to the urban soils are probably air and land traffic. Multivariate analysis was used to investigate which soil properties could influence mobility and fate of the contaminants. Whilst principal components analysis indicates carbonates and other calcium phases as probable factors controlling the dispersion of Pb in the urban soils, the linear models obtained from stepwise multiple regression analysis show that soil phosphorous (P) and manganese (Mn) are good predictors of the total soil Pb content. No robust model was obtained for the PAHs, impeding identifying environmental factors most likely to influence their dispersion in the urban soils. The solid-phase distribution study provided critical information to untangle the, at a first glance, contradictory results obtained by the multivariate analysis. Carbonates and other calcium phases, having these a probable anthropogenic origin, are soil components containing major fractions of Pb, P, and Mn.

Sewage contamination of sediments from two Portuguese Atlantic coastal systems, revealed by fecal sterols.

Fecal sterols in sediments were used to assess the degree of sewage contamination in Ria de Aveiro lagoon and Mondego River estuary for the first time. Coprostanol, the major fecal sterol, averaged 1.82 ± 4.12 µg g(-1), with maxima of 16.6 µg g(-1). The northwestern sector of the Ria and a marina at Mondego estuary showed the highest level of sewage contamination. This scenario was confirmed by several diagnostic ratios based on fecal sterols and other phytosterols. Our data revealed that in spite of the improvements achieved in the last decades, there is still a need for control the organic inputs into the aquatic environment in the studied regions.

Sources of potentially toxic elements and organic pollutants in an urban area subjected to an industrial impact.

Urban and industrial development has caused a major impact on environmental soil quality. This work assesses the extent and severity of contamination in a small urban area subjected to an industrial impact and identifies the major anthropogenic inputs. Twenty-six soil samples were collected from agricultural and urban sites, and concentrations of potentially toxic elements (As, Cd, Cu, Cr, Fe, Mn, Ni, Pb and Zn), PAHs and PCBs, were determined. In spite of the low median concentrations observed, some sites represent a potential hazard for human health and ecosystems. Concentrations of contaminants were higher than those found in a nearby city, indicating that the study area is affected by the surrounding industry. The use of multivariate statistical analyses allowed for the identification of the main factors controlling the variability of potentially toxic elements and organic pollutants in the soils. The presence of Cr, Fe, Mn and Ni was associated with geogenic inputs, and Cu, Pb, Zn, As, PAHs and PCBs were associated with anthropogenic inputs. Industry and traffic were the most important anthropogenic sources. Soil characteristics were identified as important factors controlling the spatial variability of elements, both from recognised natural and anthropogenic origin. Differences between land uses were observed, which may be attributed to both management practices and proximity to sources.

Carbofuran effects in soil nematode communities: using trait and taxonomic based approaches.

This work intends to implement the use of native soil nematode communities in ecotoxicological tests using a model pesticide and two geographically nematode communities (Mediterranean and sub-tropical) in order to obtain new perspectives on the evaluation of the toxic potential of chemical substances. The environmental condition of the nematode communities was described using a trait-based approach (grouping the organisms according to their feeding traits) and a traditional taxonomic method (identification to family level). Effects on total nematode abundance, number of families and abundance of nematode feeding groups as well as potential shifts in both trophic and family structure were assessed. Agricultural soils from Curitiba (Brazil) and Coimbra (Portugal) were sampled and the corresponding nematode communities were extracted. Part of the collected soil was defaunated and spiked with four doses of a carbofuran commercial formulation. Afterwards each of the replicates was inoculated with a nematode suspension containing ≈200 or 300 nematodes. After 14 and 28 d of exposure the nematodes were extracted, counted and identified at family level and separately classified according to their feeding traits. The patterns of nematode responses revealed a decrease in the total abundance and a reduction in the number of families. Despite the similar effects observed for both communities, statistically significant toxic effects were only found within the Portuguese community. The total nematode abundance was significantly reduced at the highest carbofuran concentrations and significant shifts in the family structure were detected. However, the trophic structure, i.e., the contribution of each feeding group for the overall community structure, did not significantly change along the contamination gradient. Results showed that using such a trait-based approach may increase the ecological relevance of toxicity data, by establishing communalities in the response to a chemical from two different taxonomic communities, although with potential loss of information on biodiversity of the communities.

Gas chromatography - optical fiber detector for assessment of fatty acids in urban soils.

Fatty acids have been used as biomarkers of the microbial community composition of soils and they are usually separated and quantified by gas-chromatography coupled to a flame ionization detector (GC-FID). The aim of this study was to develop, validate and apply a methodology based on gas chromatography coupled to optical fiber detection (GC-OF) for screening five fatty acids used as indicators of fungal and bacterial communities in urban soils. The performance of the GC-OF methodology (optical fiber detector at 1,550 nm) was evaluated by comparison with the GC-FID methodology and it was found that they were comparable in terms of linear range, detection limit and analytical errors. Besides these similar analytical characteristics, the GC-OF is much cheaper than the GC-FID methodology. Different concentrations were determined for each fatty acid indicator which in turn varied significantly between the soil samples analyzed from Lisbon ornamental gardens. Additionally, the GC-OF showed a great potential as alternative for determination of eleven or more fatty acids in urban soils.

Structural effects of the bioavailable fraction of pesticides in soil: suitability of elutriate testing.

This study focused the ecotoxicological evaluation of four different pesticides (chlorpyrifos, glyphosate, vinclozolin, endosulfan), sprayed into an agricultural soil, using a standard battery of aquatic bioassays for testing of soil elutriates: Vibrio fischeri -Microtox(®); Pseudokirchneriella subcapitata growth inhibition; Daphnia magna acute and chronic toxicity. Despite relevant pesticide residues were recovered from the soil matrix (concentrations higher than 1000 µg kg(-1)), much lower concentrations could be retrieved from elutriates (highest records for endosulfan of ca. 250 ng L(-1)and 1400 ng L(-1); dissolved and particulate concentration, respectively) and little effects were generally found in the bioassays. Lethal effects (D. magna 48 h-EC50 of 36.8%) could be noticed following exposure to the endosulfan elutriate. Elutriates induced no toxicity on V. fischeri; algal growth was generally inhibited at high elutriate dilutions and stimulated at the lower elutriate dilutions; and no overall impairment of D. magna life-history was noticed. Results revealed that cross-contamination during field application, input of organic matter and nutrients by elutriates in test solutions, and choice of test species and endpoints may constrain the ecotoxicological assessment. Suitability of established aquatic bioassay test batteries for these purposes, and questioning on whether direct assays with soil organisms could be more protective tools is discussed.

Assessment of fatty acid as a differentiator of usages of urban soils.

The determination of fatty acids (FA) has been extensively used as a sensitive and reproducible parameter for characterizing the soil microorganism communities and to detect various environmental stresses. The aim of this study was to assess the variability of FA in urban soils, in an attempt to use it as a differentiator of urban soil usage. FA were extracted from soils of five different usages (ornamental gardens, ornamental gardens/roadsides, roadsides, parks and urban agricultural areas) in three Portuguese cities and FA concentration was determined by gas chromatography coupled to mass spectrometry (GC-MS). A total of fifteen FA have been detected and the concentration of each FA, in general, decreased in the following order: Lisbon>Estarreja>Viseu, for all the usages of urban soil. According to soil microbial indicators, the concentration of Gram-negative bacteria was lower than the concentration of Gram-positive bacteria, which together were, in turn, higher than the concentration of fungi for all the usages of urban soil and city. This study assessed the FA profiles of urban soils, which differ as a function of soil usage. The FA profile also is at the source of the inference that stress in soil microorganism communities results from the different urban environment in each city.

Spatial distribution of total Hg in urban soils from an Atlantic coastal city (Aveiro, Portugal).

The aim of this study was to investigate the levels and the spatial distribution of total Hg concentrations in soils from the urban area of Aveiro (Portugal) in order to assess the impact of industrial activities and identified Hg emission sources in these urban soils. For this purpose, soils were collected in 25 sampling points (at two depths) within the urban perimeter and in places considered representative of the main green areas of the city. A median concentration of 0.091 mg kg(-1) (dry weight) was obtained, regardless the depth. Aveiro was considered a low polluted city in terms of total Hg and no direct effects of emissions of Hg from industrial activities nearby could be detected in these urban soils. Despite of the low values obtained, high variability (range of approximately 0.5 mg kg(-1)) was observed in the results. Such was considered to be related to characteristic features of soils in urban settings and to the behaviour of Hg in the urban environment.

Optimisation of a microbial bioassay for contaminated soil monitoring: bacterial inoculum standardisation and comparison with Microtox assay.

This work represents the first step to set up a toxicity testing procedure and to evaluate the sensitivity of the test microorganism to several classes of environmental pollutants. First, three different techniques were employed to standardise the microbial inoculum, then two different toxicity assessment protocols have been compared: Microtox and a dehydrogenase (DHase) activity inhibition test. The main goal was the optimisation of a microbial bioassay based on the dehydrogenase activity (DHase) inhibition in Pseudomonas fluorescens bacterial strain ATCC 13525. Triphenyl tetrazolium chloride (TTC) was used as electron acceptor and its reduction produces Triphenyl formazane (TPF). The P. fluorescens DHase inhibition bioassay was investigated for being a reliable and rapid method for assessing toxicity. The optimisation of the operating conditions resulted in a repeatable bioassay. Then, P. fluorescens and Vibrio fischeri sensitivity were firstly compared by testing Zn++, one of the reference compounds for Microtox test. In addition, other compounds (Ni++, Cd++, Cu++, phenol) were also tested with both bioassays. A high statistical significance of data was obtained with the logistic curve. The present work has demonstrated that P. fluorescens is as sensitive as Microtox culture (V. fischeri), for some of the metal ions. With reference to organic compounds, the lower sensitivity of P. fluorescens to phenol makes its use difficult in organic polluted samples.