Biogeographic responses and niche occupancy of microbial communities following long-term land-use change.

Understanding the effects of forest-to-agriculture conversion on microbial diversity has been a major goal in soil ecological studies. However, linking community assembly to the ruling ecological processes at local and regional scales remains challenging. Here, we evaluated bacterial community assembly patterns and the ecological processes governing niche specialization in a gradient of geography, seasonality, and land-use change, totaling 324 soil samples, 43 habitat characteristics (abiotic factors), and 16 metabolic and co-occurrence patterns (biotic factors), in the Brazilian Atlantic Rainforest, a subtropical biome recognized as one the world's largest and most threatened hotspots of biodiversity. Pairwise beta diversities were lower in pastures than in forest and no-till soils. Pasture communities showed a predominantly neutral model, regarding stochastic processes, with moderate dispersion, leading to biotic homogenization. Most no-till and forest microbial communities followed a niche-based model, with low rates of dispersal and weak homogenizing selection, indicating niche specialization or variable selection. Historical and evolutionary contingencies, as represented by soil type, season, and dispersal limitation were the main drivers of microbial assembly and processes at the local scale, markedly correlated with the occurrence of endemic microbes. Our results indicate that the patterns of assembly and their governing processes are dependent on the niche occupancy of the taxa evaluated (generalists or specialists). They are also more correlated with historical and evolutionary contingencies and the interactions among taxa (i.e., co-occurrence patterns) than the land-use change itself.

Biogeographic Patterns of Arbuscular Mycorrhizal Fungal Communities Along a Land-Use Intensification Gradient in the Subtropical Atlantic Forest Biome.

Information concerning arbuscular mycorrhizal (AM) fungal geographical distribution in tropical and subtropical soils from the Atlantic Forest (a global hotspot of biodiversity) are scarce and often restricted to the evaluation of richness and abundance of AM fungal species at specific ecosystems or local landscapes. In this study, we hypothesized that AM fungal diversity and community composition in subtropical soils would display fundamental differences in their geographical patterns, shaped by spatial distance and land-use change, at local and regional scales. AM fungal community composition was examined by spore-based taxonomic analysis, using soil trap cultures. Acaulospora koskei and Glomus were found as generalists, regardless of mesoregions and land uses. Other Acaulospora species were also found generalists within mesoregions. Land-use change and intensification did not influence AM fungal composition, partially rejecting our first hypothesis. We then calculated the distance-decay of similarities among pairs of AM fungal communities and the distance-decay relationship within and over mesoregions. We also performed the Mantel test and redundancy analysis to discriminate the main environmental drivers of AM fungal diversity and composition turnover. Overall, we found significant distance-decays for all land uses. We also observed a distance-decay relationship within the mesoregion scale (< 104 km) and these changes were correlated mainly to soil type (not land use), with the secondary influence of both total organic carbon and clay contents. AM fungal species distribution presented significant distance-decays, regardless of land uses, which was indicative of dispersal limitation, a stochastic neutral process. Although, we found evidence that, coupled with dispersal limitation, niche differentiation also played a role in structuring AM fungal communities, driven by long-term historical contingencies, as represented by soil type, resulting from different soil origin and mineralogy across mesoregions.

Nonylphenol causes shifts in microbial communities and nitrogen mineralization in soil microcosms.

The aims of this work was to investigate, in soil microcosms, the effects on soil microbial community structure and function of increasing concentrations of 4-Nonylphenol (NP). The lasts is a product of degradation of NPEOs (Nonylphenol polyethoxylates) with a known toxic and estrogenic capacity able to disrupt animal's hormonal systems. The effect of increasing concentrations of NP (0, 10, 30, 90, and 270 mg NP kg-1 of dry soil) in soil microcosms in three sampling dates (28, 56, and 112 days) over soil microbial activity and function were assessed. Soil microbial activity was estimated by microbial ATP content, and both bacterial and fungal communities composition were estimated using the terminal restriction fragment length polymorphism technique (T-RFLP). Abundance of ammonia-oxidizing bacteria (AOB) was estimated by qPCR of gene encoding for the bacterial ammonia-monoxygenase (amoA). Changes in biologically mediated soil properties were also assessed, namely water-soluble NH+4, NO-2 and NO-3 content, the two last allowing the assessment of mineralization rates. NP-spiking had some unexpected impacts on microbial community structure and functions, since (i) impacted both bacterial and fungal communities structure at the highest NP concentration tested, bacterial communities were resistant to lower concentrations, while fungal communities were increasingly impacted until the end of the incubation at day 112; (ii) no community structure resilience was observed in bacteria at the highest NP concentration nor for fungi at any concentration; (iii) microbial activity decreased with NP after 28 and 56 d, but increased in the last sampling at the highest concentrations tests, coupled to an enrichment in AOB taxa after 56 and 112 days, that at least partly explain also explain the observed speed up of nitrification rates.

Disentangling the effects of the aqueous matrix on the potential toxicity of liquid pig manure in sub-tropical soils under semi-field conditions.

Inadequate application of liquid pig manure (LPM) may pose risks to the soil due to the potential contaminants that exists, as well as by the large water input that can originate excessive moisture. By using Terrestrial Model Ecosystems, this study aimed to evaluate the effect of application of LPM (82% moisture) using the application rates of 20, 50 and 150 m3 ha-1 and also of the corresponding amount of water to understand the origin of effects on the soil fauna of two sub-tropical soils. In general, the results obtained for the two soil types indicated that LPM (150 m3 ha-1) changed the composition of soil fauna, with an increase in the abundance of insect larvae and dipterans, but a decrease in the number of earthworms and enchytraeids. Microbial biomass, soil respiration and the nutrients Mg, K and P increased with the amount of LPM supplied to the soil. When analysing the effects of adding large volumes of water via the LPM, results showed that application did not originate significant effects on the parameters measured. The differences obtained when comparing both treatments were mainly attributed to the organic load brought by the LPM and not to the amount of water without LPM.

Environmental risk assessment of pesticides in tropical terrestrial ecosystems: Test procedures, current status and future perspectives.

Despite the increasing use of pesticides in tropical countries, research and legislative efforts have focused on their temperate counterparts. This paper presents a review of the literature on environmental risk assessment of pesticides for tropical terrestrial agroecosystems. It aims at evaluating potential differences in pesticide risk between temperate and tropical regions as well as to highlight research needs in the latter. Peculiarities of pesticide risks in tropical terrestrial agroecosystems are discussed in subsections 1) agricultural practices; 2) research efforts; 3) fate and exposure; 4) toxicity testing methods; and 5) sensitivity. The intensive and often inadequate pesticide application practices in tropical areas are likely to result in a relatively greater pesticide exposure in edge-of-field water bodies. Since pesticide fate may be different under tropical conditions, tropical scenarios for models estimating predicted environmental pesticide concentrations should be developed. Sensitivity comparisons do not indicate a consistent similar, greater or lower relative sensitivity of tropical soil organisms as compared to temperate organisms. However, several methods and procedures for application in the tropics need to be developed, which include: 1) identifying and collecting natural soils to be used as reference test substrates in tests; 2) identifying and discerning the range of sensitivity of native test species to soil contaminants; 3) developing test guidelines applicable to tropical/subtropical conditions; and 4) developing methods and procedures for higher tier testing for full development and implementation of environmental risk assessment schemes.

Ecotoxicological effects of fipronil, neem cake and neem extract in edaphic organisms from tropical soil.

Veterinary medicines are widely applied for the treatment and prevention of animal diseases. Consequently, animal manure contains significant amounts of environmental pollutants that are potential sources of environmental pollution when inappropriately applied in soils. This work aimed to evaluate ecotoxicological effects of doses of commercial fipronil, neem cake and neem extract in the survival and reproduction of earthworms (Eisenia andrei), enchytraeidae (Enchytraeus crypticus) and springtails (Folsomia candida) in Oxisol and tropical artificial soil (TAS). Applications of fipronil, neem cake and extract in soil were carried out according to standardized ISO methodologies by using a random experimental design with five replicates. Toxic effects of fipronil for springtails in Oxisol and TAS were observed with LC50 of 0.26 mg kg-1 (0.18-0.35 mg kg-1) and 0.29 mg kg-1 (0.22-0.37 mg kg-1), respectively. It was not observed significant toxic effects of fipronil for earthworms and enchytraeidae in both soils. However, significant amounts of juvenile earthworm and adult enchytraeidae decreased in fipronil doses higher than 10 mg kg-1. Neem cake and extract were not toxic for earthworms and enchytraeidae but, significant amounts of juvenile springtails decreased in neem cake doses from 500 to 1000 mg kg-1. It can be concluded that the use of veterinary medicines containing synthetic compounds for preventing diseases in animals needs to be controlled to avoid environmental pollution after applying manure in soil. Veterinary medicines containing natural compounds as neem cake and extract are eco-friendly and could be efficiently applied in soil in a sustainable way.

Placing arbuscular mycorrhizal fungi on the risk assessment test battery of plant protection products (PPPs).

Arbuscular mycorrhizal fungi (AMF) are mutualistic symbionts considered a key group in soil systems involved in the provision of several ecosystem services. Recently they have been listed by EFSA as organisms to be included in the test battery for the risk assessment of plant protection product (PPPs). This study aimed to contribute to improve the ISO Protocol (ISO 10832: 2009) by assessing the feasibility of using other AMF species under different test conditions. Overall, results showed that AMF species Gigaspora albida and Rhizophagus clarus (selected out of five AMF species) are suitable to be used in spore germination tests using the ISO protocol (14 days incubation with sand or artificial soil as substrate) to test PPPs. However, several modifications to the protocol were made in order to accommodate the use of the tested isolates, namely the incubation temperature (28 °C instead of 24 °C) and the change of reference substance (boric acid instead of cadmium nitrate). The need for these changes, plus the results obtained with the three fungicides tested (chlorothalonil, mancozeb and metalaxyl-M) and comparisons made with literature on the relevance of the origin of AMF isolates in dictating the adequate test conditions, emphasize the importance of adjusting test conditions (AMF species/isolates and test temperature) when assessing effects for prospective risk assessment targeting different climatic zones. So, further studies should be conducted with different AMF species and isolates from different climatic regions, in order to better define which species/isolate and test conditions should be used to assess effects of a particular PPP targeting a given climatic zone.

A TME study with the fungicide pyrimethanil combined with different moisture regimes: effects on enchytraeids.

Today's ecosystems are influenced by different factors that could evolve into stressors. Effects of pesticides, especially in agricultural areas, may interact with environmental factors, such as soil moisture fluctuation caused by global climate change. In this contribution, two semi-field studies conducted in Germany and Portugal with terrestrial model ecosystems are presented. Their aim was to assess the effects of the fungicide pyrimethanil under different soil moisture levels on Enchytraeidae. In Portugal a no observed effect concentration design was chosen, using two concentration levels: the maximum application rate (MAR) according to the safe use registration within the European Union and five times the MAR (1.82 and 9.09 mg/kg dry soil, respectively). Both concentrations did neither affect the total enchytraeid abundance nor single populations. In Germany an ECx design (effect concentration) was conducted, using 11 concentrations. In general, 14 EC50 values for different combinations of single species, moisture level and sampling date were determined. The strongest effects were found in dry soil, particularly for Fridericia connata (EC50: 3.48 mg/kg dry soil after 8 weeks of exposure). The advantages and challenges of these test designs are discussed with regard to the registration process of pesticides in the European Union. In any case, enchytraeids are suitable test organisms in such higher tier studies for the combined evaluation of chemical and climatic stressors due to their usually high diversity and abundances and their close contact with the soil solution.

Ecotoxicological assessment of a dredged sediment using bioassays with three species of soil invertebrates.

The ecotoxicity of a dredged sediment from the Guanabara Bay (Rio de Janeiro, RJ, Brazil) was evaluated using reproduction tests with Eisenia andrei, Folsomia candida and Enchytraeus crypticus, and avoidance and feeding inhibition tests with Folsomia candida. The sediment was mixed with artificial soil to obtain the following doses: 1.25, 2.5, 5.0, 10.0, 20.0 and 40.0 %. Lead, nickel, chromium, copper and zinc concentrations were determined in the test mixtures. In reproduction tests, E. andrei was the most sensitive species (EC50 = 2.94 %), followed by F. candida (EC50 = 7.72 %) and E. crypticus (EC50 = 10.10 %). The percentage of initial weight of earthworms was significantly higher in all test concentrations compared to the control except at the highest one where earthworms biomass significantly decreased. No feeding inhibition of F. candida was observed for any test mixture and the number of organisms with a dark gut (the fed collembolans) generally increased with the increasing dose of sediment. Significant avoidance responses of F. candida were observed towards all test mixtures, however, the avoidance behaviour was the less sensitive endpoint after feeding inhibition. The results showed that chemical analysis is not sufficient to foresee toxic effects in terrestrial systems resulting from sediment disposal in soil if not complemented with an ecotoxicological evaluation.

Seed dressing pesticides on springtails in two ecotoxicological laboratory tests.

Terrestrial ecotoxicological tests are powerful tools for assessing the ecological risks that pesticides pose to soil invertebrates, but they are rarely used to evaluate seed dressing pesticides. This study investigated the effects of seed dressing pesticides on survival and reproduction of Folsomia candida (Collembola), using standardized ecotoxicological tests (after ISO guidelines with few adaptations for tropical conditions). Commercial formulations of five seed dressing pesticides were tested individually in Tropical Artificial Soil (TAS): the insecticides imidacloprid, fipronil, thiametoxam, and the fungicides captan and carboxin+thiram. Thiametoxam, captan, and carboxin+thiram were only lethal to F. candida at the highest concentration tested (1000mg of active ingredient kg(-1) of dry soil). Imidacloprid and fipronil were lethal at lower concentrations (100 and 10mg a.i. kg(-1) soil d.w, respectively), however, these concentrations were much higher than those predicted (PEC) for soil. Imidacloprid and fipronil were the most toxic pesticides in both tests, reducing significantly collembolan reproduction (EC20=0.02 and 0.12mga.i.kg(-1) soil d.w, respectively). Further studies under more realistic conditions are needed, since imidacloprid and fipronil reduced collembolan reproduction at concentrations below or close to their respective PECs.

Ethoprophos fate on soil-water interface and effects on non-target terrestrial and aquatic biota under Mediterranean crop-based scenarios.

The present study aimed to assess the environmental fate of the insecticide and nematicide ethoprophos in the soil-water interface following the pesticide application in simulated maize and potato crops under Mediterranean agricultural conditions, particularly of irrigation. Focus was given to the soil-water transfer pathways (leaching and runoff), to the pesticide transport in soil between pesticide application (crop row) and non-application areas (between crop rows), as well as to toxic effects of the various matrices on terrestrial and aquatic biota. A semi-field methodology mimicking a "worst-case" ethoprophos application (twice the recommended dosage for maize and potato crops: 100% concentration v/v) in agricultural field situations was used, in order to mimic a possible misuse by the farmer under realistic conditions. A rainfall was simulated under a slope of 20° for both crop-based scenarios. Soil and water samples were collected for the analysis of pesticide residues. Ecotoxicity of soil and aquatic samples was assessed by performing lethal and sublethal bioassays with organisms from different trophic levels: the collembolan Folsomia candida, the earthworm Eisenia andrei and the cladoceran Daphnia magna. Although the majority of ethoprophos sorbed to the soil application area, pesticide concentrations were detected in all water matrices illustrating pesticide transfer pathways of water contamination between environmental compartments. Leaching to groundwater proved to be an important transfer pathway of ethoprophos under both crop-based scenarios, as it resulted in high pesticide concentration in leachates from Maize (130µgL(-1)) and Potato (630µgL(-1)) crop scenarios, respectively. Ethoprophos application at the Potato crop scenario caused more toxic effects on terrestrial and aquatic biota than at the Maize scenario at the recommended dosage and lower concentrations. In both crop-based scenarios, ethoprophos moved with the irrigation water flow to the soil between the crop rows where no pesticide was applied, causing toxic effects on terrestrial organisms. The two simulated agricultural crop-based scenarios had the merit to illustrate the importance of transfer pathways of pesticides from soil to groundwater through leaching and from crop rows to the surrounding soil areas in a soil-water interface environment, which is representative for irrigated agricultural crops under Mediterranean conditions.

Disposal of dredged sediments in tropical soils: ecotoxicological effects on earthworms.

The upper limit concentrations of metals established by international legislations for dredged sediment disposal and soil quality do not take into consideration the properties of tropical soils (generally submitted to more intense weathering processes) on metal availability and ecotoxicity. Aiming to perform an evaluation on the suitability of these threshold values in tropical regions, the ecotoxicity of metal-contaminated dredged sediment from the Guanabara Bay (Rio de Janeiro, Brazil) was investigated. Acute and avoidance tests with Eisenia andrei were performed with mixtures of dredged sediment with a ferralsol (0.00, 6.66, 13.12, 19.98, and 33.30 %) and a chernosol (0.00, 6.58, 13.16, 19.74, and 32.90 %). Mercury, lead, nickel, chromium, copper, and zinc concentrations were measured in test mixtures and in tissues of surviving earthworms from the acute tests. While ferralsol test mixtures provoked significant earthworm avoidance response at concentrations ≥13.31 %, the chernosol mixtures showed significant avoidance behavior only at the 19.74 % concentration. The acute tests showed higher toxicity in ferralsol mixtures (LC50 = 9.9 %) compared to chernosol mixtures (LC50 = 16.5 %), and biomass increased at the lowest sediment doses in treatments of both test soils. Most probably, the expansive clay minerals present in chernosol contributed to reduce metal availability in chernosol mixtures, and consequently, the ecotoxicity of these treatments. The bioconcentration factors (BCF) for zinc and copper were lower with increasing concentrations of the dredged sediment, indicating the existence of internal regulating processes. Although the BCF for mercury also decreased with the increasing test concentrations, the known no biological function of this metal in the earthworms metabolism lead to suppose that Hg measured was not present in bioaccumulable forms. BCFs estimated for the other metals were generally higher in the highest dredged sediment doses.

Quantitative assessments of honeybee colony's response to an artificial vibrational pulse resulting in non-invasive measurements of colony's overall mobility and restfulness.

In this work we aim to provide a quantitative method allowing the probing of the physiological status of honeybee colonies by providing them with a gentle, short, external artificial vibrational shockwave, and recording their response. The knock is provided by an external electromagnetic shaker attached to the outer wall of a hive, driven by a computer with a 0.1 s long, monochromatic vibration at 340Hz set to an amplitude that occasionally yields a mild response from the bees, recorded by an accelerometer placed in the middle of the central frame of the colony. To avoid habituation, the stimulus is supplied at randomised times, approximately every hour. The method is pioneered with a pilot study on a single colony hosted indoors, then extended onto eight outdoors colonies. The results show that we can quantitatively sense the colony's overall mobility, independently from another physiological aspect, which is phenomenologically explored. Using this, a colony that is queenless is easily discriminated from the others.

Assessment of the reproduction of six collembolan species in tropical soils naturally rich in potentially toxic elements.

Laboratory ecotoxicological tests are important tools for the management of environmental changes derived from anthropogenic activities. Folsomia candida is usually the model species used in some procedures. However, this species may not be sufficiently representative of the sensitivity of the other collembolan species. This study aimed to evaluate (i) the effects of soils naturally rich in potentially toxic elements (PTE) and soil characteristics on the reproduction and survival of different collembolan species, (ii) whether the habitat function of these soils is compromised, and (iii) to what extent F. candida is representative of the other collembolan species. For this, reproduction tests with six collembolan species were conducted in 14 different samples of soils. In general, collembolan reproduction was not completely inhibited in none of the natural tested soils. Even soils with high pollution load index values did not negatively affect collembolan reproduction for most of the species. In contrast, the lowest collembolan reproduction rates were found in a visually dense soil (lowest volume/weight ratio), highlighting that soil attributes other than total PTE concentration also interfere in the reproduction of collembolans. Our results support the idea that the F. candida species might not be representative of other collembolan species and that laboratory tests to assess soil contaminations should be conducted using diverse collembolan species.

Allele Frequencies of Genetic Variants Associated with Varroa Drone Brood Resistance (DBR) in Apis mellifera Subspecies across the European Continent.

Implementation of marker-assisted selection (MAS) in modern beekeeping would improve sustainability, especially in breeding programs aiming for resilience against the parasitic mite Varroa destructor. Selecting honey bee colonies for natural resistance traits, such as brood-intrinsic suppression of varroa mite reproduction, reduces the use of chemical acaricides while respecting local adaptation. In 2019, eight genomic variants associated with varroa non-reproduction in drone brood were discovered in a single colony from the Amsterdam Water Dune population in the Netherlands. Recently, a new study tested the applicability of these eight genetic variants for the same phenotype on a population-wide scale in Flanders, Belgium. As the properties of some variants varied between the two studies, one hypothesized that the difference in genetic ancestry of the sampled colonies may underly these contribution shifts. In order to frame this, we determined the allele frequencies of the eight genetic variants in more than 360 Apis mellifera colonies across the European continent and found that variant type allele frequencies of these variants are primarily related to the A. mellifera subspecies or phylogenetic honey bee lineage. Our results confirm that population-specific genetic markers should always be evaluated in a new population prior to using them in MAS programs.

Bridging the Gap between Field Experiments and Machine Learning: The EC H2020 B-GOOD Project as a Case Study towards Automated Predictive Health Monitoring of Honey Bee Colonies.

Honey bee colonies have great societal and economic importance. The main challenge that beekeepers face is keeping bee colonies healthy under ever-changing environmental conditions. In the past two decades, beekeepers that manage colonies of Western honey bees (Apis mellifera) have become increasingly concerned by the presence of parasites and pathogens affecting the bees, the reduction in pollen and nectar availability, and the colonies' exposure to pesticides, among others. Hence, beekeepers need to know the health condition of their colonies and how to keep them alive and thriving, which creates a need for a new holistic data collection method to harmonize the flow of information from various sources that can be linked at the colony level for different health determinants, such as bee colony, environmental, socioeconomic, and genetic statuses. For this purpose, we have developed and implemented the B-GOOD (Giving Beekeeping Guidance by computational-assisted Decision Making) project as a case study to categorize the colony's health condition and find a Health Status Index (HSI). Using a 3-tier setup guided by work plans and standardized protocols, we have collected data from inside the colonies (amount of brood, disease load, honey harvest, etc.) and from their environment (floral resource availability). Most of the project's data was automatically collected by the BEEP Base Sensor System. This continuous stream of data served as the basis to determine and validate an algorithm to calculate the HSI using machine learning. In this article, we share our insights on this holistic methodology and also highlight the importance of using a standardized data language to increase the compatibility between different current and future studies. We argue that the combined management of big data will be an essential building block in the development of targeted guidance for beekeepers and for the future of sustainable beekeeping.

Functional Groups Response to Water Deficit in Mediterranean Ecosystems.

Enhanced drought, more frequent rainfall events and increased inter-annual variability of precipitation are the main trends of climate expected for the Mediterranean. Drought is one of the most important stressors for plants and significantly impacts plant communities causing changes in plant composition and species dominance. Through an experiment under controlled conditions, we assessed the response of Mediterranean species from different functional groups (annual grass, annual forb, annual legume, and perennial shrub) to moderate and severe water deficit. Changes in plant traits (leaf dry matter), biomass and physiology (water status, photosynthesis, pigments, and carbohydrate) were evaluated. The studied species differed in their response to water deficit. Ornithopus compressus, the legume, showed the strongest response, particularly under severe conditions, decreasing leaf relative water content (RWC), pigments and carbohydrates. The grass, Agrostis pourreti and the forb, Tolpis barbata, maintained RWC, indicating a higher ability to cope with water deficit. Finally, the shrub, Cistus salviifolius, had the lowest response to stress, showing a higher ability to cope with water deficit. Despite different responses, plant biomass was negatively affected by severe water deficit in all species. These data provide background for predicting plant diversity and species composition of Mediterranean grasslands and Montado under climate change conditions.

The effect of COVID-19 confinement on the activity behaviour of red deer.

The COVID-19 pandemic has drastically affected people's social habits, especially those related to outdoor activities. We intended to understand the effects of the two national lockdowns in Portugal on the presence and activity of a wild population of red deer (Cervus elaphus) by analysing data from camera traps installed at Lousã mountain, in the central part of Portugal. The cameras were set between 2019 and 2021, and a total of 2434 individual contacts of red deer and 182 contacts of people were recorded. Results showed a higher human presence in the mountain area during the COVID-19 outbreak, especially during the first lockdown in 2020 (0.05 ± 0.17 individuals/day), compared to the same period of the year before the pandemic (0.02 ± 0.05 individuals/day), which resulted in an increase of people by 150%. The increase in human presence did not have a significant direct effect on the presence of red deer. Despite the low overlap of activity patterns between people and red deer, deer showed avoidance behaviour in the 24 h after the detection of human presence on camera traps, as well as an increase in daily activity during the 2020 lockdown, showing red deer's awareness of human visitation. These results showed that people's increased search for cultural services in wild environments during COVID-19 lockdowns, such as hiking and biking, seemed to influence the population of red deer, albeit momentarily.

Soil pH matters in the ecotoxicity of Basamid® to freshwater microalgae and macrophytes.

Intensive agriculture along with the use of agrochemicals has been associated with low soil fertility, soil erosion, and soil acidity. Management of soil pH through liming is a common practice in agriculture to increase soil fertility and nutrient availability. When altering soil pH, different chemical reactions occur depending on soil composition and agrochemicals presence. Basamid® is a fumigant used worldwide targeting soil nematodes, fungi, and weeds in diverse crops, that can reach freshwater ecosystems by leaching through the soil layers. The major goal of this work was to assess the influence of soil pH in the toxicity of Basamid® eluates to the microalgae Raphidocelis subcapitata and the duckweed Lemna minor. For this, eluates were prepared from soils with different pH (5.5, 6.5 and 7.5), contaminated with the recommended dose of Basamid® corresponding to 145.7 mg of dazomet/Kg soil. Soil was amended with calcium carbonate (CaCO3). Raphidocelis subcapitata and L. minor were exposed to the eluates during 72 h and 7 days respectively, and multiple endpoints were assessed: growth rate, biomass, pigment as chlorophyl content and cell damage. Results showed that soil pH can influence the performance of the tested species and also be a major factor in influencing Basamid®'s toxicity. However, a clear pattern of the influence of soil pH on Basamid®'s toxicity was not observed and was species dependent. For R. subcapitata lower soil pHs induced higher toxicity of Basamid®'s to the algae [ED50 for growth rate: 30 % (confidence limits-CL: 22.8-37.2) for soil pH 5.5; >100 % for soil pH 6.5 and pH 7.5], while for L. minor the opposite was observed [ED50 for number of fronds: 27.2 % (CL: 22.8-31.6) for pH 5.5; 20.3 % (CL: 10.0-30.6) for pH 6.5 and 10.7 % (CL: 6.3-15.1)]. Overall, these results showed that leachates of Basamid® through soils, at recommended doses, can have a severe impact on aquatic systems, with or without the influence of abiotic factors.

Ecotoxicity of eluates obtained from Basamid® contaminated soils is pH dependent: A study with Hydra viridissima, Xenopus laevis and Danio rerio.

Agrochemicals are mostly used to deplete pests and treat diseases in terrestrial agro-ecosystems. However, their transport through the soil, by leaching and/or runoff, may cause them to reach aquatic systems. Environmental parameters, such as soil pH, can affect this transport, by influencing the magnitude of agrochemicals degradation and chemical reaction. This work aimed at investigating the influence of soil pH on the toxicity of eluates obtained from Basamid® contaminated soils to Hydra viridissima, Xenopus laevis and Danio rerio. For this, a natural soil with pH amended to 5.5, 6.5 and 7.5, was spiked with the recommended dose (RD) of Basamid® (145 mg dazomet/kg soil) and eluates (Ba-E) were prepared with the respective species culture medium. Dilutions of the eluates (0.14-100%), obtained from the three soils (Ba-E 5.5, Ba-E 6.5 and Ba-E 7.5, corresponding to soil spiked with Basamid® RD at soil pH of 5.5, 6.5 and 7.5, respectively), were used to expose the organisms. Results showed that for H. viridissima increased soil alkalinity provoked less mortality comparatively to lower soil pH [LD50,96h of Ba-E 5.5: 10.6% and LD50,96h of Ba-E 7.5: 21.2%]. As for X. laevis and D. rerio Ba-E lethal ecotoxicity was similar across soil pH (LD50,96h varied from 5.7 to 6.9% and from 2.1 to 4.3%, respectively). For malformations, 20% effect dilution (ED) in H. viridissima was significantly higher at Ba-E 7.5 (ED20,96h: 17.4%), comparatively to Ba-E 5.5 and Ba-E 6.5 (ED20,96h: 7.9% and 7.7%, respectively). From the three tested organisms and based on both lethal and sublethal effects, H. viridissima presented the highest tolerance to Basamid® eluates and soil pH was a major factor determining the fumigant toxicity, with higher soil pH levels inducing, lower toxicity. The eluates obtained from soils contaminated with RD of Basamid® induced severe effects to the three aquatic species.