Hotspots of soil pollution: Possible glyphosate and aminomethylphosphonic acid risks on terrestrial ecosystems and human health.

The study presents a literature review of glyphosate (GLY) occurrence and its breakdown product, aminomethylphosphonic acid (AMPA), in soils worldwide, but with a specific focus on South America. In addition, an ecological risk approach based on the ecotoxicological endpoints for key soil biota (e.g., collembolans, and earthworms) assessed the impact of GLY and AMPA on these organisms. A generic probabilistic model for human health risk was also calculated for the different world regions. For what reports the risk for edaphic species and the level of pollution under the worst-case scenario, the South American continent was identified as the region of most concern. Nonetheless, other areas may also be in danger, but no risk could be calculated due to the lack of data. Since tropical countries are the top food exporters worldwide, the results obtained in this study must be carefully examined for their implications on a global scale. Some of the factors behind the high levels of these two chemicals in soils are debated (e.g., permissive protection policies, the extensive use of genetically modified crops), and some possible guidelines are presented that include, for example, further environmental characterisation and management of pesticide residues. The present review integrates data that can be used as a base by policymakers and decision-makers to develop and implement environmental policies.

Earthworm communities in long-term no-tillage systems and secondary forest fragments in Paraná, Southern Brazil.

The area destined for agricultural production in Paraná state in Brazil is ~6 million hectares, of which 79% are under no-tillage systems (NTS) that can positively affect earthworm populations. Furthermore, earthworm abundance and richness can be valuable soil quality. This study assessed earthworm communities in long-term no-tillage sites (NTS) and nearby secondary Atlantic Forest (SF) fragments. Sampling was performed in June 2018 and May 2019 using the quantitative Tropical Soil Biology and Fertility handsorting method (ISO-TSBF), complemented by qualitative sampling in three municipalities: Faxinal, Mauá da Serra and Palmeira, in NTS and SF. Eleven earthworm species, belonging to five families (Benhamiidae, Glossoscolecidae, Megascolecidae, Ocnerodrilidae, and Rhinodrilidae) were found along with seven native species (Glossoscolex sp.22, Glossoscolex sp.23, Fimoscolex sp.21, Fimoscolex sp.24, Fimoscolex sp.42, Andiorrhinus duseni and Urobenus brasiliensis) and four exotic or cosmopolitan species (Dichogaster gracilis, Amynthas gracilis, Metaphire californica and Pontoscolex corethrurus). The cosmopolitan P. corethrurus dominated Faxinal and Mauá da Serra, while in Palmeira Fimoscolex and Glossoscolex were the most abundant. Six species belonging to Glossoscolex and Fimoscolex were new to science and must be described. Overall, 239 individuals were found. In 2018 126 individuals were found (76 in NTS and 50 in SF) and in 2019, only 112 individuals were found (45 in NTS and 67 in SF). The highest earthworm abundance was in Faxinal (123 individuals), with 76 individuals in NTS and 47 in SF. In Mauá da Serra the same pattern was observed (SF>NTS), while in Palmeira in overall, fewer individuals (38) were collected (24 in NTS and 14 in SF). The NTS at Faxinal and SF Mauá da Serra in 2019 had only/more exotic/cosmopolitan species, while at the other dates and sites >50% of the specimens sampled were native species. Overall, forests had more native species than exotics: in 2018, natives represented > 75% of all specimens, but in 2019 both Faxinal and Mauá da Serra had more exotic species (> 65%), while at Palmeira, they represented 50% of the individuals. Earthworm total biomass for forest sites was higher in Faxinal (3.2 g) and NTS in Mauá da Serra (1.7 g) in 2019. Overall, the total biomass was higher in SF sites than NTS. Using a classification available for earthworm populations in no-tillage systems, all three sites were considered to have poor quality in terms of abundance (ind m-2), while for species richness, they were considered to have moderate (Faxinal and Mauá da Serra) and good quality (Palmeira).

Metabolic Consequences of the Water We Drink: A Study Based on Field Evidence and Animal Model Experimentation.

The effect of the chronic consumption of water contaminated with residual concentrations of DDT's metabolites (DDD-dichlorodiphenyldichloroethane and DDE-dichlorodiphenyldichloroethylene) found in the environment were evaluated on the biometric, hematological and antioxidant system parameters of the hepatic, muscular, renal and nervous tissues of Wistar rats. The results showed that the studied concentrations (0.002 mg.L-1 of DDD plus 0.005 mg.L-1 of DDE) could not cause significant changes in the hematological parameters. However, the tissues showed significant alteration in the activity of the antioxidant system represented by the increase in the activity of the enzymes gluthathione S-transferases in the liver, superoxide dismutase in the kidney, gluthathione peroxidase in the brain, and several changes in enzymatic activity in muscle (SOD, GPx and LPO). The enzymes alanine aminotransaminase (ALT) and aspartate aminotransaminase (AST) were also evaluated for the amino acids' metabolism in the liver, with ALT showing a significant increase in the exposed animals. In the integrative analysis of biomarkers (Permanova and PCOA), the studied concentrations showed possible metabolic changes and damage to cellular structures evidenced by increased oxidative stress and body weight gain among the treated animals. This study highlights the need for further studies on the impact of banned pesticides still present in soils that may induce adverse effects in organisms that may prevail in future generations and the environment.

Effects of 2,4,6-Trichlorophenol on Clarias batrachus: a biomarkers approach.

2,4,6-Trichlorophenol (2,4,6-TCP) is a common waste among the resulting chlorophenols generated in the production of common products classified as an extremely toxic, mutagenic, carcinogenic and highly persistent xenobiotic in the environment. To evaluate the impact of 2,4,6-TCP in aquatic systems, the catfish species Clarias batrachus has been selected to test its toxicity due to its high market value and consumption in India. Here is presented the impact of this compound on different physiological parameters of fish: haematological parameters (haemoglobin, total erythrocyte count, total leucocyte count and mean corpuscular haemoglobin), biochemical parameters (total serum protein and total serum glucose), growth and reproductive parameters (condition factor, hepatosomatic index, maturity index, specific growth rate, growth hormone, 17ß-estradiol and testosterone), exposed to two concentrations of 2,4,6-TCP (0.5 mg/L and 1 mg/L - 1/10th and 1/20th of the LC50) for a period of 15, 30 and 45 days. The results showed that C. batrachus even when exposed to the lower concentration (0.5 mg/L) for the shortest time (15 days) negatively impacted the organism in all the assessed parameters. This was highlighted by the Integrated Biomarker Response index (IBR), showing worse scores for the treatments (up to 20 × worse than the control). This work highlights the importance of continued research on the impact of 2,4,6-TCP, on an important commercial, supported by the high environmental persistence of this compound that can reach the same range of tested concentrations.

Pesticides in a case study on no-tillage farming systems and surrounding forest patches in Brazil.

With the growing global concern on pesticide management, the relationship between its environmental recalcitrance, food security and human health has never been more relevant. Pesticides residues are known to cause significant environmental contamination. Here, we present a case study on long-term no-tillage farming systems in Brazil, where Glyphosate (GLY) has been applied for more than 35 years. GLY and its main breakdown product, aminomethylphosphonic acid (AMPA) were determined in topsoil (0-10 cm) samples from no-tillage fields and nearby subtropical secondary forests by high-performance liquid chromatography coupled with a fluorescence detector. In addition, the presence of carbamates, organochlorines, organophosphates and triazines were also screened for. GLY and AMPA were present in all soil samples, reaching values higher than those described for soils so far in the literature. A significant decrease for AMPA was observed only between the secondary forest and the farm's middle slope for site B. GLY and AMPA were observed respectively at peak concentrations of 66.38 and 26.03 mg/kg soil. GLY was strongly associated with forest soil properties, while AMPA associated more with no-tillage soil properties. Soil texture was a significant factor contributing to discrimination of the results as clay and sand contents affect GLY and AMPA retention in soils. This was the first study to report DDT and metabolites in consolidated no-tillage soils in Brazil (a pesticide fully banned since 2009). Based on human risk assessment conducted herein and the potential risk of GLY to local soil communities, this study offers a baseline for future studies on potential adverse effects on soil biota, and mechanistic studies.

Unravelling the molecular mechanisms of nickel in woodlice.

During the last few years, there has been an alarming increase in the amount of nickel (Ni) being released into the environment, primarily due to its use in the production of stainless steel but also from other sources such as batteries manufacturing and consequent disposal. The established biotic ligand models provide precise estimates for Ni bioavailability, in contrast, studies describing the mechanisms underpinning toxicological effect of Ni are scarce. This study exploits RNA-seq to determine the transcriptomic responses of isopods using Porcellionides pruinosus as an example of a terrestrial metal-resistant woodlouse. Furthermore, the recently proposed model for Ni adverse outcome pathways (Ni-AOP) presents an unprecedented opportunity to fit isopod responses to Ni toxicity and define Porcellionides pruinosus as a metalomic model. Prior to this study, P. pruinosus represented an important environmental sentinel, though lacking genetic/omic data. The reference transcriptome generated here thus represents a major advance and a novel resource. A detailed annotation of the transcripts obtained is presented together with the homology to genes/gene products from Metazoan and Arthropoda phylum, Gene Ontology (GO) classification, clusters of orthologous groups (COG) and assignment to KEGG metabolic pathways. The differential gene expression comparison was determined in response to nickel (Ni) exposure and used to derive the enriched pathways and processes. It revealed a significant impact on ion trafficking and storage, oxidative stress, neurotoxicity, reproduction impairment, genetics and epigenetics. Many of the processes observed support the current Ni-AOP although the data highlights that the current model can be improved by including epigenetic endpoints, which represents key chronic risks under a scenario of Ni toxicity.

Mercury accumulation from food decreases collembolans' growth.

In the terrestrial environment, mercury (Hg) contamination can be originated from different inorganic and metal-organic sources, redistributed and transformed in soils. In the present study, the effects of contaminated food with environmentally relevant concentrations of Hg were evaluated in the soil-dwelling invertebrate Folsomia candida. Changes in growth rate and Hg bioaccumulation levels were observed at different concentrations of Hg in food, which can be complementary for data already available for reproduction and survival from standardized protocols. Collembolan growth was recorded every two days, and their growth rate along with a Von Bertalanffy's growth curve were derived showing that growth was dependent on Hg food concentration. Also, the final length of animals reflected the Hg concentration in food, with differences in all treatments comparing to non-exposed organisms. Toxicokinetic patterns from different Hg concentrations in food were not significantly different during the uptake phase, but differences were found in the depuration phase. Combining the two approaches, collembolans seem to invest their energy for depuration processes, neglecting other vital processes, such as growth. Also, contaminated food avoidance possibly occurred, thus decreasing their feeding and contaminant intake. Therefore, growth tests in collembolans can act as complementary tools to bioaccumulation and reproductive assays, towards a mechanistic understanding of how organisms use their energy upon contamination. Changes in growth rate, even at low and environmentally relevant concentrations, could be a warning signal when occurring in species with key roles in ecosystems. Also, this study highlights the importance of these complementary tests for a better and complete approach to risk assessment studies.

Multigenerational effects of carbendazim in Daphnia magna: From a subcellular to a population level.

Anthropogenic activities such as the use of pesticides may affect aquatic biota populations, due to potential agricultural runoffs or disposals. Carbendazim is one example of a widely used fungicide with a high potential to end up in aquatic ecosystems through runoff. Deleterious effects observed at the individual level are possibly explained by changes in homeostasis at the cellular level, and both factors can then be used to predict effects at the population level. In the present study, an isoclonal population of Daphnia magna (clone K6) was exposed to a concentration that mimics relevant levels of carbendazim in the environment over 12 generations. The effects of carbendazim were assessed in some generations using the following endpoints: biochemical biomarkers (cholinesterase, catalase, and glutathione-S-transferase), lipid peroxidation and energy-related parameters (carbohydrates, lipids, and proteins along with available energy and energy consumption), parental longevity, and population growth (r). Long-term exposure to carbendazim had no effect on the intrinsic rate of natural increase (r) of adult D. magna, but longevity was decreased at the F12 generation compared to that of control. Differences between the exposed and nonexposed populations were found for cholinesterase, glutathione-S-transferase, and lipid peroxidation. However, for catalase and energy-related parameters, no differences were observed between these 2 populations. Natural variability was seen throughout the test period, under control conditions, within the 12 generations. Overall, carbendazim induced some effects at the subcellular level that translated into changes in longevity but these later vanished in terms of population effects. Environ Toxicol Chem 2019;38:412-422. © 2018 SETAC.

Joint effects of chlorpyrifos and mancozeb on the terrestrial isopod Porcellionides pruinosus: A multiple biomarker approach.

The exposure to pesticides by nontarget soil biota has long been regarded as a serious downside of modern agricultural regimes and the subject of heated debate. Of utmost relevance is the exposure to pesticide mixtures because their effects have been shown to not necessarily reflect the individual toxicity of their components, and even the simple addition of effects may lead to consequences not clearly anticipated. In the present study, a multiple biomarker approach was employed to identify the mechanistic and time effects underlying several single and mixture treatments of chlorpyrifos (CPF) and mancozeb (MCZ) in juveniles and adults of the terrestrial isopod Porcellionides pruinosus. The effects of the individual pesticides and the mixtures at recommended doses were mostly transitory under these controlled conditions and one-pulse exposure. Whereas imbalances were identified on detoxification and oxidative stress-related enzymes, the isopods generally showed the ability to recover through the end of the experiment. However, juveniles displayed greater vulnerability than adults. Most of the differences between life stages occurred in energy-related parameters where distinct performances and stress-handling behaviors were observed, suggesting higher metabolic costs to juveniles. Our results stress that understanding the time dependence of the underlying mechanisms governing the joint effects of the pesticides can help in assessing and anticipating the effects of the pesticide mixtures. Moreover, we emphasize the importance of taking life stage-related differences into consideration when evaluating the environmental risks of pesticides and pesticide mixtures. Environ Toxicol Chem 2018;37:1446-1457. © 2018 SETAC.

In field conditions, commercial pigment grade TiO2 was not harmful to terrestrial isopods but reduced leaf litter fragmentation.

We investigated the effects of a commercial pigment grade rutile TiO2 on the terrestrial isopod Porcellio scaber in three locations that differed in terms of abiotic and biotic conditions: the laboratory, open air, and the closed barn. Mortality and isopod energy reserves (digestive gland total proteins, lipids and carbohydrates) were not affected following 14days exposure to up to 1000mg TiO2 per kg dry leaves (mg/kg) under any experimental scenario. However, in the field tests, isopods consumption of TiO2-coated leaves was reduced compared to that of uncoated leaves and the decrease was not dose-dependent. The highest reduction was in the closed barn (45-56%) rather than in the open-air (38-40%). In laboratory-based food choice tests, isopods neither preferred nor avoided leaves coated with TiO2, suggesting that rather than sensing the TiO2 on the leaves directly, the isopods under open-air and barn exposure responded to altered attractiveness and/or palatability of the TiO2 amended leaves. We propose that this could be due to altered microbial population on the leaves, a hypothesis that requires further investigation. Although short-term exposure to atmospheric deposition of up to 1000mg/kg commercial TiO2 is unlikely to pose an immediate threat to isopod mortality and energy balance, reduced leaf feeding may have implications for the decomposition of plant material.

Toxicity interaction between chlorpyrifos, mancozeb and soil moisture to the terrestrial isopod Porcellionides pruinosus.

A main source of uncertainty currently associated with environmental risk assessment of chemicals is the poor understanding of the influence of environmental factors on the toxicity of xenobiotics. Aiming to reduce this uncertainty, here we evaluate the joint-effects of two pesticides (chlorpyrifos and mancozeb) on the terrestrial isopod Porcellionides pruinosus under different soil moisture regimes. A full factorial design, including three treatments of each pesticide and an untreated control, were performed under different soil moisture regimes: 25%, 50%, and 75% WHC. Our results showed that soil moisture had no effects on isopods survival, at the levels assessed in this experiment, neither regarding single pesticides nor mixture treatments. Additivity was always the most parsimonious result when both pesticides were present. Oppositely, both feeding activity and biomass change showed a higher sensitivity to soil moisture, with isopods generally showing worse performance when exposed to pesticides and dry or moist conditions. Most of the significant differences between soil moisture regimes were found in single pesticide treatments, yet different responses to mixtures could still be distinguished depending on the soil moisture assessed. This study shows that while soil moisture has the potential to influence the effects of the pesticide mixture itself, such effects might become less important in a context of complex combinations of stressors, as the major contribution comes from its individual interaction with each pesticide. Finally, the implications of our results are discussed in light of the current state of environmental risk assessment procedures and some future perspectives are advanced.

Metabolic responses of the isopod Porcellionides pruinosus to nickel exposure assessed by (1)H NMR metabolomics.

UNLABELLED: This work aimed at characterizing the metabolome of the isopod Porcellionides pruinosus and at assessing its variations over 14 days under laboratory culture conditions and upon exposure to the contaminant metal Nickel (Ni). The spectral profiles obtained by (1)H NMR spectroscopy were thoroughly assigned and subjected to multivariate analysis in order to highlight consistent changes. Over 50 metabolites could be identified, providing considerable new knowledge on the metabolome of these model organisms. Several metabolites changed non-linearly with Ni dose and exposure time, showing distinct variation patterns for initial (4 days) and later time points (7 and 14 days). In particular, at day 4, several amino acids were increased and sugars were decreased (compared to controls), whereas these variations were inverted for longer exposure, possibly reflecting earlier and more intensive moulting. Other variations, namely in betaines and choline-containing compounds, were suggested to relate with osmoregulation and detoxification mechanisms. Ni also had a marked effect on several nucleotides (increased upon exposure) and a moderate impact on lipids (decreased upon exposure). Overall, this study has provided new information on the Ni-induced metabolic adaptations of the P. pruinosus isopod, paving the way for improved mechanistic understanding of how these model organisms handle soil contamination. SIGNIFICANCE: This study provided, for the first time to our knowledge, a detailed picture of the NMR-detectable metabolome of terrestrial isopods and of its fluctuations in time and upon exposure to the contaminant metal Nickel. Several time- and dose-dependent changes were highlighted, providing mechanistic insight into how these important model organisms handle Ni contamination.

Long-term exposure of the isopod Porcellionides pruinosus to nickel: Costs in the energy budget and detoxification enzymes.

Terrestrial isopods from the species Porcellionides pruinosus were exposed to the maximum allowed nickel concentration in the Canadian framework guideline (50 mg Ni/kg soil) and to 5× this concentration (250 mg Ni/kg soil). The exposure lasted for 28 days and was followed by a recovery period of 14 days where organisms were changed to clean soil. Organisms were sampled after 24 h, 48 h, 96 h, 7 days, 14 days, 21 days, and 28 days of exposure, and at days 35 and 42 during the recovery period. For each sampling time the acetylcholinesterase (AChE), glutathione-S-transferases (GST), catalase (CAT), lactate dehydrogenase (LDH) activities were determined as well as lipid peroxidation rate (LPO) along with lipids, carbohydrates, proteins content, energy available (Ea), energy consumption (Ec) and cellular energy allocation (CEA). The integrated biomarker response (IBR) was calculated for each sampling time as well as for each one of the above parameters. In addition, mortality was also recorded throughout the assay. The results obtained showed that nickel induced oxidative stress, evidenced by results on GST, GPx, CAT or LPO, but also on changes in the energy reserves content of these organisms. In addition, this study showed that these organisms possess a specific strategy to handle nickel toxicity. In this case, biomarkers were associated with costs in the energy budget, and the increase of energy reserves has a compensation for that cost.

Suitability of enzymatic markers to assess the environmental condition of natural populations of Gambusia affinis and Daphnia magna--a case study.

In recent years, the use of biochemical markers, especially in the assessment of toxic effects and modes of action, under controlled laboratory conditions has increased. However, transposing their use to in situ monitoring or risk assessment evaluations has encountered barriers, mainly related to the difficulty in interpreting the meaning of biochemical variation. In this work, we aimed at understanding if biochemical marker activities (cholinesterase, glutathione S-transferase and lactate dehydrogenase) can be used to monitor the health status of natural populations of fish (Gambusia affinis) and daphnids (Daphnia magna). For that, two ponds with different water properties were chosen as study sites, and organisms collected at four sampling periods along the year. The pattern of biochemical marker responses was not the same in the two species, showing higher integrated biochemical marker response values in the winter for G. affinis and in the autumn for D. magna, suggesting specificities that must be taken into account in biomonitoring programmes by including representative species of several trophic levels. In the case of G. affinis, the differences in key physicochemical parameters between the two ponds (especially dissolved oxygen levels) did not seem to affect biochemical marker levels as if organisms were already perfectly adapted to their environment. In general, seasonal variation of water quality seems to have an important role on biochemical marker responses. Several parameters above Environmental Quality Standards were identified such as dissolved oxygen (DO), ammonia, nitrites, sulphides and metals, but eventual responses to these stressors could not be discriminated from natural variation except for particular cases.

Biomarkers and energy reserves in the isopod Porcellionides pruinosus: the effects of long-term exposure to dimethoate.

Terrestrial isopods from the species Porcellionides pruinosus were exposed to the recommended field dose application (0.4 mg/kg soil) and a sublethal concentration (10mg/kg soil) of dimethoate at two temperatures that can be generally found in several countries (20°C and 25°C) and are commonly used as reference temperatures. The organisms were exposed for 28 days and sampled at the following time points: 24h, 48 h, 96 h, 7 days, 14 days, 21 days, 28 days; organisms were then changed to clean soil for a recovery period of 14 days during which organisms were sampled on day 35 and 42. For each sampling time, the enzyme activities of acetylcholinesterase (AChE), glutathione-S-transferases (GST), catalase (CAT), lactate dehydrogenase (LDH) were determined as well as the following: total lipid, carbohydrate and protein content; energy available (Ea); energy consumption (Ec); cellular energy allocation (CEA) and lipid peroxidation rate (LPO). The integrated biomarker response (IBR) was calculated for each sampling time and for each of the above parameters. Mortality was also recorded during the study. The results obtained showed that dimethoate causes toxicity by several mechanisms. This study found evidence for the inhibition of the acetylcholinesterase enzyme, which has been previously reported, and also evidence of oxidative stress, which altered the levels of GST, CAT or LPO. In addition, the study showed that the two concentrations used of dimethoate caused the activation of different general detoxification mechanisms, and also that the same concentration at different temperatures induced different toxicity responses.

Environmental- and growth stage-related differences in the susceptibility of terrestrial isopods to UV radiation.

Global environmental changes are nowadays one of the most important issues affecting terrestrial ecosystems. One of its most significant expressions is the increasing ultraviolet radiation (UVR) arising from the human-induced depletion in ozone layer. Therefore, to investigate the effects of UVR on the terrestrial isopod Porcellionides pruinosus a multiple biomarker approach was carried out. Two experiments were performed in order to analyze the importance of the exposure environment and the growth stage on the UV-induced damages. First, adult individuals were exposed to UVR in three exposure environments (soil, soil with leaves, and plaster). Thereafter, three growth stages using soil as the exposure condition were tested. Integrated biomarker responses (IBR) suggested that UV effects were higher in plaster, and mostly identified by changes in acetylcholinesterase and glutathione-S-transferases activities, lipid peroxidation rates, and total energy available. The effects in soil and soil with leaves were not so clear. In the growth stages' experiment, juveniles and pre-adults were found to be more affected than adults, with the greatest differences between irradiated and non-irradiated isopods occurring in energy-related parameters. Our findings suggest that soil surface-living macrofauna may be prone to deleterious effects caused by UVR, highlighting the importance of taking the media of exposure and growth stage in account.

Preliminary evaluation of the toxic effects of the antifouling biocide Sea-Nine 211™ in the soft coral Sarcophyton cf. glaucum (Octocorallia, Alcyonacea) based on PAM fluorometry and biomarkers.

Sea-Nine 211™ is a new biocide specifically formulated for antifouling paints and being considered to have a low environmental impact. Even with a short environmental half-life, this compound can cause toxic effects on marine organisms. This study used PAM fluorometry and biomarkers of oxidative stress (GST, CAT and LPO) to monitor potential toxic effects of Sea-Nine 211™ on fragments of the soft coral Sarcophyton cf. glaucum. After exposure to concentrations of 1-100 µg l(-1) for 72 h, CAT activity was inhibited under the two highest concentrations, being in accordance with the activity of GST. LPO activity (as TBARS) and photosynthetic efficiency of endosymbiotic zooxanthellae were not significantly affected. These results show that PAM fluorometry alone cannot detect the full effects of Sea-Nine 211™ on Sarcophyton cf. glaucum and should be used together with other biomarkers. This holobiont driven approach to evaluate chemical toxicity in photosynthetic corals is therefore recommended for biocides which are not photosystem II inhibitors.