Ecotoxicity of multiple stressors on soil organisms, assessment of the risk of pollutants accounting for landscape variability and application of bioremediation technologies to sewage sludge deposition: in vivo and in silico approaches

297 p.[EN]The anthropic activities during the last decade are causing damages on ecosystems. In fact, animal husbandry, agriculture, and industrial activities have increased the proportion of soils contaminated with pesticides and metals. The presence of pollutants could affect soil organisms and the Ecosystem Services provided; thus, a proper risk evaluation is required in order to assess the final environmental impacts of pollutants. In silico models are tools designed to predict environmental concentrations of released pollutants that could assist on substances evaluation and regulation. Meanwhile, the health paradigm, understanding the natural environment as an interrelated compartment, has been spread within the scientific community. Hence, the measurement of toxicological effects produced by contaminants on soil organisms (using in vivo tests and biomarkers) gained a high reputation for evaluating environmental health. This work presents different approaches, techniques, software, guidelines and tests for assessing contaminated soils; crucial for an integrative soil assessment.[EU]Azken hamarkadako jarduera antropikoek ekosistemetan desorekak sortzen-hasiak dira. Horrela, abeltzaintzak, nekazaritzak eta aktibitate industrialek pestizida eta-metalekin kutsaturiko lurzoruen proportzioa emendarazi dute. Kutsatzaileen-presentziak, lurzoruetan bizi diren organismoak eta beraz, lurzoruak eskaintzen dituen-Zerbitzu Ekosistemikoak erasatea ekarri dezake, arrisku ebaluazio egokien beharra-eskatuz. In silico modeloak, kutsatzaile baten aplikazio osteko ingurumen-kontzentrazioak aurreikusteko tresna egokiak dira, substantzien arrisku ebaluazioan eta-erregulazioan lagun dezaketenak. Bestetik, kutsatzaileak lurzoruan duten inpaktua-ebaluatze aldera, osasunaren paradigma ikuspuntua; hots, medio naturalak intererlazionatutako-konpartimentu gisa ulertzen dituena, hedatzen hasi da komunitate-zientifikoaren baitan. Era honetan, kutsatzaileek lurzoru organimoengan eragindako-efektu toxikologikoen neurketak (in vivo test-ak eta biomarkatzaileak erabiltzen-dituztenak) omen handia irabazi dute ingurumenaren osasuna ebaluatzerako orduan.-Lan honetan, kutsatutako lurzoruak ebaluatzeko ikuspuntu, teknika, software, gidalerro-eta test ezberdinak aurkezten dira; lurzoru ebaluazio integral bat burutzeko-beharrezkoak direnak.This work was funded by EFSA European Food Safety Authority (NP/EFSA/AFSCO/2016/03); MINECO (CTM2017- 87766-R), Basque Government (Grupos Consolidados Tipo A. Refs. IT-810-13; IT-1302-19; and IHOBE-EcoInnovacion BIOREM17-); Gernika-Lumo Town Hall; UPV/EHU (PES17/10; 20/45)

Impacts of sewage sludges deposition on agricultural soils: Effects upon model soil organisms

During years sewage sludges have been worldwide poured in agricultural soils to enhance vegetal production. The "Landfill 17" located in Gernika-Lumo town (43°19'28.9"N 2°40'30.9"W) received for decades sewage sludges from the local Waste Water Treatment Plant (WWTP) with agricultural purposes. To this WWTP, several pollutants as heavy metals (Cd, Cr, Ni, Pb), PAHs (benzo(a)pyrene among many others) and pesticides (i.e. dieldrin) could have arrived from local industry and be widespread all over the landfill. Soil invertebrates like earthworms and plants are of special interest due to their close contact with the polluted matrix and their potential effects by the presence of pollutants. In this context, the aim of the present work was to determine the health status of landfill soils by evaluating the effects on model soil organisms exerted by long-lasted pollutants after on site deposition of WWTP active sludges. With such a purpose, different standard toxicity tests and cellular level endpoints were performed on lettuce and earthworms. Indeed, germination (EPA 850.4100) and root elongation (EPA 850.4230) tests were carried out in Lactuca sativa, while OECD acute toxicity test (OECD-204), reproduction test (OECD-222) and Calcein-AM viability test with coelomocytes were applied in Eisenia fetida worms. For the exposure, soils collected in the landfield containing low, medium and high concentrations of pollutants were selected, and as reference LUFA 2.3 natural standard soil was chosen. While no differences were shown in the assays with L. sativa, significant differences between sludge exposed groups and control group were recorded with E. fetida, with lower coelomocyte number and viability and higher tissue metal accumulation after 28days of exposure to polluted soils. These results confirmed the impact of contaminants to soil biota even after long periods of time.Basque Government (IT810-13), CTM2017-87766-R from MINECO, PhytoSUDOE-SOE1/P5/E0189, and Gernika City Hall

Predicting environmental concentrations and the potential risk of Plant Protection Products (PPP) on non-target soil organisms accounting for regional and landscape ecological variability in european soils

[EN] Plant Protection Products (PPP) raise concerns as their application may cause effects on some soil organisms considered non-target species which could be highly sensitive to some pesticides. The European Food and Safety Authority (EFSA), in collaboration with the Joint Research Centre (JRC) of the European Commission, has developed guidance and a software tool, Persistence in Soil Analytical Model (PERSAM), for conducting soil exposure assessments. EFSA PPR Panel has published recommendations for the risk assessment of non-target soil organisms. We have used PERSAM for calculating PPPs predicted environmental concentrations (PECs); and used the estimated PEC for assessing potential risks using Toxicity Exposure Ratios (TER) for selected soil organisms and good agricultural practices. Soil characteristics and environmental variables change along a latitudinal axis through the European continent, influencing the availability of PPP, their toxicity upon soil biota, and hence, impacting on the risk characterization. Although PERSAM includes as input geographical information, the in-formation is aggregated and not further detailed in the model outputs. Therefore, there is a need to develop landscape based environmental risk assessment methods addressing regional variability. The objective was to integrate spatially explicit exposure (PECs) and effect data (biological endpoints i.e. LC50, NOEC, etc.) to estimate the risk quotient (TER) of four PPP active substances (esfenvalerate, cyclaniliprole, picoxystrobin, fenamidone) on non-target species accounting European landscape and agricultural variability. The study was focused on the effects produced by the above-mentioned pesticides on two soil organisms: E. fetida earthworms and Folsomia sp. collembolans. After running PERSAM assuming a worst case application of PPPs, PECs in total soil and pore water were obtained for different depths in northern, central and southern European soils. With this data, soil variability and climatic differences among soils divided in three large Euroregions along a latitudinal transect (Northern, Central, Southern Europe) were analysed. Summarising, a trend to accumulate higher PECs and TERs in total soil was observed in the north decreasing towards the south. Higher PECs and TERs could be expected in pore water in southern soils, decreasing towards the north. The risk disparity between pollutant concentrations at different soils compartments should be taken into account for regulatory purposes, as well as the potential landscape variabilities among different Euroregions.This work has been partially funded by European Food Safety Authority (EFSA) through the grant NP/EFSA/AFSCO/2016/03. The authors Miguel Santos and Jose V Tarazona are employed with the EFSA in the Pesticides Peer Review and the Scientific Committee and Emerging Risks Units. However, the present article is published under the sole responsibility of the authors and may not be considered as an EFSA scientific output. The positions and opinions presented in this article are those of the authors alone and do not represent the views of EFSA

Environmental risk assessment of PPP application in European soils and potential ecosystem service losses considering impacts on non-target organisms

The use of Plant Protection Products (PPPs) is leading to high exposure scenarios with potential risk to soil organisms, including non-target species. Assessment of the effects of PPPs on non-target organisms is one of the most important components of environmental risk assessment (ERA) since they play crucial functions in ecosystems, being main driving forces in different soil processes. As part of the framework, EFSA is proposing the use of the ecosystem services approach for setting specific protection goals. In fact, the services provided by soil organisms can be impacted by the misuse of PPPs in agroecosystems. The aim of this work was to assess PPPs potential risk upon ecosystem services along European soils, considering impacts on earthworms and collembola. Four well-known (2 insecticides-esfenvalerate and cyclaniliprole- and 2 fungicides - picoxystrobin and fenamidone-) worst case application (highest recommended application) were studied; exploring approaches for linked observed effects with impacts on ecosystem services, accounting for their mode of action (MoA), predicted exposure, time-course effects in Eisenia fetida and Folsomia sp. and landscape variability. The selected fungicides exerted more effects than insecticides on E. fetida, whereas few effects were reported for both pesticides regarding Folsomia sp. The most impacted ecosystem services after PPP application to crops appeared to be habitat provision, soil formation and retention, nutrient cycling, biodiversity, erosion regulation, soil remediation/waste treatment and pest and disease regulation. The main factors to be taken into account for a correct PPP use management in crops are discussed.This work has been partially funded by European Food Safety Authority (EFSA) through the grant NP/EFSA/AFSCO/2016/03. The author Miguel J.G. Santos is employed with the EFSA in the PREV Unit. The author Jose V Tarazona was employed by EFSA until October 2022. The present article is published under the sole responsibility of the au thors and may not be considered as an EFSA scientific output. The po sitions and opinions presented in this article are those of the authors alone and do not represent the views of EFSA.S

Sedimentuen toxikotasunaren azterketa integrala ekologikoki adierazgarriak diren espezieak erabiliz

Euskal kostaldean eta, nagusiki, estuarioetan, sedimentu ugari aurkitzen ditugu, egungo zein iraganeko giza jardueren eraginez substantzia toxiko ugari metatzen dituztenak. Horrek ingurumen-arazoak dakartza, eta sedimentu horiek modu egokian kudeatzera bultzatu behar gaitu. Horretarako, kutsatzaileok sedimentuetan duten portaera ulertu behar da, bai eta sedimentuen karakterizazio sakona egin ere. Gaur egunean indarrean dagoen legediak zenbait gidalerro eskaintzen dituen arren, hainbat lanek gidalerro horien mugak agerian utzi dituzte. Bereziki garrantzitsua da ohiko neurketa kimikoak sedimentuetan bizi diren organismoen ikerketekin uztartzea. Izan ere, haietan burutzen diren entseguek informazio oso baliagarria eskain dezakete sedimentuen toxikotasunaren eta, beraz, haien kudeaketa-beharraren inguruan. Entsegu horietarako erabiltzen diren bizidunek sedimentuan bizitzea eta han funtzio ekologiko garrantzitsua izatea komeni da. Horien artean Hediste diversicolor poliketoa dugu, sedimentuen kutsaduraren ebaluaziorako eta bioentsegu zehatz eta fidagarriak burutzeko morganismo aproposa dena. Horiez gain, in vitro teknika egokiak eta sentikorrak garatu dira espezie honetan, eta horrela esperimentazioan erabiltzen diren bizidunen kopurua mugatu egiten da. Beraz, gure sedimentuetan metatutako kutsatzaileen maila altuek izan ditzaketen arriskuen ebaluazio fidagarria lor daiteke, baldin eta analisi kimikoak eta ekologikoki adierazgarriak diren espezieetan gertatutako efektuak (analisi ekotoxikologikoak) integratzen badira.; In the Basque coast and mainly in estuaries, sediment deposition and accumulation occurs. It is noteworthy that different human activities have influenced on sediments that have undergone important inputs of different toxic substances over the years and even today. Since polluted sediments could pose a risk for the environment, we should proceed properly for an accurate environmental management. For that, firstly it is necessary to study and understand the behavior of pollutants in sediments and then to carry out characterization of sediments. Although the legislation currently in force provides certain guidelines, several works have revealed the limitations of these policies. Therefore, in addition to conventional chemical measurements, organisms living in estuarine sediments and tests performed in them can provide very useful information on sediment toxicity (and thus future use and management). Mainly, if these sentinel organisms are ecologically relevant species that inhabit those niches. Among organisms living in sediment, the polychaete Hediste diversicolor has been used for sediment toxicity assessment using bioassays. Moreover, in vitro techniques have been described in this species as very useful tools, limiting the number of organisms used in experiments and showing them as sensitive tools. Therefore, a reliable assessment of the hazards resulting from high levels of pollutants in sediments can be achieved, after integrating chemical analysis and effects measured in ecologically relevant species (ecotoxicological analysis)

Gentle remediation options for soil with mixed chromium (VI) and lindane pollution: biostimulation, bioaugmentation, phytoremediation and vermiremediation

Gentle Remediation Options (GROs), such as biostimulation, bioaugmentation, phytoremediation and vermiremediation, are cost-effective and environmentally-friendly solutions for soils simultaneously polluted with organic and inorganic compounds. This study assessed the individual and combined effectiveness of GROs in recovering the health of a soil artificially polluted with hexavalent chromium [Cr(VI)] and lindane. A greenhouse experiment was performed using organically-amended vs. non-amended mixed polluted soils. All soils received the following treatments: (i) no treatment; (ii) bioaugmentation with an actinobacteria consortium; (iii) vermiremediation with Eisenia fetida; (iv) phytoremediation with Brassica napus; (v) bioaugmentation + vermiremediation; (vi) bioaugmentation + phytoremediation; and (vii) bioaugmentation + vermiremediation + phytoremediation. Soil health recovery was determined based on Cr(VI) and lindane concentrations, microbial properties and toxicity bioassays with plants and worms. Cr(VI) pollution caused high toxicity, but some GROs were able to partly recover soil health: (i) the organic amendment decreased Cr(VI) concentrations, alleviating toxicity; (ii) the actinobacteria consortium was effective at removing both Cr(VI) and lindane; (iii) B. napus and E. fetida had a positive effect on the removal of pollutants and improved microbial properties. The combination of the organic amendment, B. napus, E. fetida and the actinobacteria consortium was the most effective strategy.Fil: Lacalle, Rafael G.. Universidad del País Vasco; EspañaFil: Aparicio, Juan Daniel. Universidad Nacional de Tucumán; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Artetxe, Unai. Universidad del País Vasco; EspañaFil: Urionabarrenetxea, Erik. Universidad del País Vasco; EspañaFil: Polti, Marta Alejandra. Universidad Nacional de Tucumán; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Soto, Manuel. Universidad del País Vasco; EspañaFil: Garbisu, Carlos. Centro de Investigación. Neiker - Tecnalia; EspañaFil: Becerril, José M.. Universidad del País Vasco; Españ

Innovative in vivo and in vitro bioassays for the establishment of toxicity thresholds of pollutants in sediment quality assessment using polychaetes and their immune cells.

Sediment toxicity testing has become a crucial component for assessing the risks posed by contaminated sediments and for the development of sediment quality assessment strategies. Commonly used organisms for bioassays with estuarine sediments include amphipods, Arenicola marina polychaetes and echinoids. Among the latter, the Sea Urchin Embryo test (SET) is the most widely used. However, one relevant limitation of this bioassay is the unavailability of gametes all year-round, particularly outside the natural spawning seasons. Consequently, the establishment of an appropriate and complementary model organism for a continuous assessment of sediment quality is recommended. A reliable assessment of the hazards resulting from pollutants in sediments or pore water, can be achieved with ecologically relevant species of sediment such as the polychaete Hediste diversicolor, which is widespread in estuaries and has the capacity to accumulate pollutants. The aim of this work was to develop reliable in vivo and in vitro bioassays with H. diversicolor and its coelomocytes (immune cells) to determine the toxicity thresholds of different contaminants bounded to sediments or resuspended into water. Polychaetes were exposed to sublethal concentrations of CuCl2 (in vivo) and a non-invasive method for collection of polychaetes coelomocytes was applied for the in vitro bioassay, exposing cells to a series of CuCl2 and AgNPs concentrations. Same reference toxicants were used to expose Paracentrotus lividus following the SET (ICES N 51; Beiras et al., 2012) and obtained toxicity thresholds were compared between the two species. In vivo exposure of polychaetes to high concentrations of Cu produced weight loss and histopathological alterations. After in vitro approaches, a significant decrease in coelomocytes viability was recorded for both toxicants, in a monotonic dose-response curve, at very short-exposure times (2h). The toxicity thresholds obtained with polychaetes were in line with the ones obtained with the SET, concluding that their sensitivity is similar. In conclusion, in vivo and in vitro bioassays developed with H. diversicolor are accurate toxicity screenings of pollutants that could be bounded to sediments or dissolved in the pore water, and may complement the SET outside the spawning period of the echinoderms. The bioassays herein developed could be applied not only to establish the toxicity thresholds of individual compounds or mixtures, but also to assess the toxicity of field collected sediments.The authors thank Basque Government (Grant to Consolidated Research Groups; IT1302-19, IT1213-19) and Spanish Ministry of Economy and Competitiveness (SEAdimenTOX project, CTM 2017-87766-R)

Lurzoru kutsatuen karakterizazio intentsiboa in vivo eta in silico fokatzeak erabiliz

The anthropic activities during the last decade are causing damages on ecosystems. In fact, animal husbandry, agriculture, and industrial activities have increased the proportion of soils contaminated with pesticides and metals. The presence of pollutants could affect soil organisms and the Ecosystem Services provided; thus, a proper risk evaluation is required in order to assess the final environmental impacts of pollutants. In silico models are tools designed to predict environmental concentrations of released pollutants that could assist on substances evaluation and regulation. Meanwhile, the health paradigm, understanding the natural environment as an inter-related compartment, has been spread within the scientific community. Hence, the measurement of toxicological effects produced by contaminants on soil organisms (using in vivo tests and biomarkers) gained a high reputation for evaluating environmental health. This work presents different approaches, techniques, software, guidelines and tests for assessing contaminated soils; crucial for an integrative soil assessment; Azken hamarkadatako jarduera antropikoak ekosistemetan desorekak sortzen hasiak dira. Horrela, abeltzaintzak, nekazaritzak eta aktibitate industrialek pestizida eta metalekin kutsaturiko lurzoruen proportzioa emendarazi dute. Kutsatzaileen presentziak, lurzoruetan bizi diren organismoak eta beraz, lurzoruak eskaintzen dituen Zerbitzu Ekosistemikoak erasatea ekarri dezake, arrisku ebaluazio egokien beharra eskatuz. In silico modeloak, kutsatzaile baten aplikazio osteko ingurumen kontzentrazioak aurreikusteko tresna egokiak dira, substantzien arrisku ebaluazioan eta erregulazioan lagun dezaketenak. Bestetik, kutsatzaileak lurzoruan duten inpaktua ebaluatze aldera, osasunaren paradigma ikuspuntua; hots, medio naturalak inter-erlazionatutako konpartimentu gisa ulertzen dituena, hedatzen hasi da komunitate zientifikoaren baitan. Era honetan, kutsatzaileek lurzoru organimoengan eragindako efektu toxikologikoen neurketak (in vivo test-ak eta biomarkatzaileak erabiltzen dituztenak) omen handia irabazi dute ingurumenaren osasuna ebaluatzerako orduan. Lan honetan, kutsatutako lurzoruak ebaluatzeko ikuspuntu, teknika, software, gidalerro eta test ezberdinak aurkezten dira; lurzoru ebaluazio integral bat burutzeko beharrezkoak direnak

Araztegi-lokatzak jasotako lurzoruaren analisi toxikologikoa zizare eta landareak erabiliz

Urtetik urtera areagotzen diren giza populazioak eta jarduera industrialak zabortegi, isuri-puntu eta betetze guneen emendioa eragin du. Zabortegi horien artean, 48046-00181 kodearekin, "17-zabortegia" dago, Gernika-Lumon kokatua (43°19’28,9”N 2°40’30,9”W.). Zabortegi horretan, Gernikako araztegiko arazketa-lokatzak isuri izan ziren ongarri gisa urteetan zehar; eta, horrela, Gernikako industrian sortutako hainbat kutsatzaile (metal astunak, PAHak, pestizidak, etab.) bertan lurperatu ziren. Isurketaren eraginez gehien kaltetutako espezieen artean, azpimarratzekoak dira landareak eta zizareak edo antzeko lurzoruko ornogabeak; batez ere, berorien eta lurzoru-matrizearen arteko kontaktu estuagatik. Testuinguru horretan, lan honen helburua da lokatz horien isurketak eragindako efektuen ebaluazio bat gauzatzea, lurzoruari beste erabilera bat emate aldera; eta, horretarako, zabalki ezagunak diren test estandarizatu eta biomarkatzaileak erabiliko dira: organismo (uraza eta zizare), denbora (3, 7, 28 eta 56 egunera) eta konplexutasun biologiko desberdinetan (zelula-mailatik populazio-mailara arte). Besteak beste, OECD-204 (toxikotasun akutuaren testa), OECD-222 (ugalketa-testa) eta kaltzeina AM bideragarritasun-testak aplikatu ziren Eisenia fetida zizarean, paraleloki, Lactuca sativa landareetan ernetze- eta elongazio-testak gauzatu ziren heinean. Azterketa toxikologikoen ostean, kontrolarekiko desberdintasun esanguratsuak ikusi ziren: erauzitako zelomozito kantitatean, bideragarritasun zelularrean, ehunetan metatutako metal-kontzentrazioetan, hazkuntza-parametroetan (pisu-galeran) eta ugalketa-parametroetan, kutsatzaileek lurzoru biotan eragindako afekzioa berretsiz. L. sativa espeziarekin egindako frogetan, aldiz, ez zen desberdintasun nabarmenik preziatu talde esperimentalen artean.; Increasing global population, along with a rising industrial activity and soil scarceness led to landfill, spill and filling point extension. Among this landfills, “Landfill 17” can be found with 48046-00181 code; located in Gernika-Lumo (43°19'28,9"N 2°40'30,9"W, Basque Country). There, sewage sludges coming from Gernika waste water treatment plant were poured with a hose with agricultural purposes; thus, many pollutants with industrial origin (heavy metals, PAHs and pesticides among others) ended up in the mentioned soils. Among the species mostly affected by the contamination, plants and earthworms must be highlighted due to their close relation with soil matrix; and therefore, soil pollution. In this context, the aim of this work is to evaluate the effects exerted by pollutants in the spilling in order to assess potential Landfill future uses; using for that widely known biomarkers and standardized tests in different organisms (lettuce and earthworms), exposure times (3, 7, 28 and 56 days) and complexity levels (from cellular level to populational level) for that. Indeed, OECD-204 (acute toxicity test), OECD-222 (reproduction test) and calcein AM tests were applied in Eisenia fetida earthworms; while, germination and elongation tests were applied on Lactuca sativa plants. After carrying out ecotoxicological assays, significant differences respect to the control were observed in: extruded coelomocyte quantity, cell viability, tissue metal accumulations, growth (weight loss) parameters or reproductive parameters; proving pollutants affection upon soil biota. However, no significant differences were observed between experimental groups in those tests carried out with L. sativa

Aldibereko teknika biologikoen bitartezko lurzoru kutsatuen erremediazioa

Anthropogenic activities led to the proliferation of polluted sites or potentially polluted sites around the world; including Basque Country. Technologies to remediate or clean these soils, mainly polluted by heavy metals or organic pollutants are diverse and of different nature. On the one hand, classic physicochemical remediation techniques can be found; with high yields and short application times; but, high costs and high ecological impacts. On the other hand, biological techniques could be found. These technologies are cheaper and more environmental-friendly; but, they require long application times. Among most used bioremediation technologies plant-based phytoremediation, worm based vermiremediation, and mibrobia based bioremediation (or bioaugmentation) can be found. This paper deals with the main features of these techniques and their applicability; both as single remediadion techniques or combined.; Gaur egun ugariak dira mundu mailan zein Euskal Herrian jarduera antropikoek kutsatuta dauden edo kutsatuta egon daitezkeen lurrak. Metal astun edo elementu organikoekin kutsatuta dauden lurzoru horiek erremediatzeko teknikak anitzak eta izaera ezberdinekoak dira. Alde batetik, erremediazio-teknika fisiko-kimiko klasikoak daude, errendimendu altu eta aplikazio-denbora laburrak dituztenak, baina, kontrara, kostu altu eta inpaktu ekologiko handiak dituztenak. Bestetik, teknika biologikoak daude, merkeagoak eta ingurumenarekiko adeitsuagoak, baina, aplikazio-denbora luzeak behar dituztenak. Horien artean daude landareekin gauzatzen den fitoerremediazioa, zizareekin gauzatzen den bermierremediazioa zein mikrobioekin gauzatzen den bioerremediazioa. Lan honetan, teknika horien ezaugarri nagusiak eta aplikagarritasuna jorratzen dira, bakarkako erremediazio-teknika zein teknika konbinatu gisa