Ecotoxicity of single and mixture of perfluoroalkyl substances (PFOS and PFOA) in soils to the earthworm Aporrectodea caliginosa.

Per and polyfluoroalkyl substances (PFAS) are persistent compounds that are massively used in industry, consumer goods and fire-fighting foams. Soil contamination by PFAS is a major environmental concern, and there is a lack of knowledge on both their ecotoxicological mechanisms and the concentrations that induce adverse effects especially to non-target organisms, particularly in the case of PFAS mixtures. This study contributes to filling these gaps by assessing and modelling the effects of PFAS (in single and in mixtures for PFOS and PFOA at different environmental doses) on juvenile endogeic earthworms of a common species in European soils (Aporrectodea caliginosa) at different levels of biological organization (sub-individual and individual). The results showed for the first time combined strong ecotoxicological effects of PFAS on earthworm survival, integumental integrity, growth, sexual maturity and on genomic stability notably with the induction of DNA breaks associated with no abnormal oxidative DNA-lesion levels. Our results demonstrated significant effects at 0.3 mg kg-1 and additive effects in case of mixtures.

Influence of copper treatment on bioaccumulation, survival, behavior, and fecundity in the fruit fly Drosophila melanogaster: Toxicity of copper oxide nanoparticles differ from dissolved copper.

Copper oxide (CuO) NPs are widely used and subsequently released into terrestrial ecosystems. In the present study, bioaccumulation and effects of CuO NPs and dissolved Cu was examined in the fruit fly Drosophila melanogaster after 7 and 10 days dietary exposure at concentrations ranging between 0.09 and 1.2 mg Cu ml-1 for dissolved Cu and between 0.2 and 11 mg Cu ml-1 for CuO NPs. Both Cu forms were bioaccumulated and affected survival and climbing in flies, but not egg-to-adult development. Dissolved Cu caused higher mortality than CuO NPs (CuO NPs 10-days LC50 was 2 times higher), whereas NPs affected climbing and decreased the number of eggs laid per female, potentially affecting fruit fly population size in terrestrial environments. Thus, the study indicates that CuO NPs might cause effects that are different from dissolved Cu due to differences in the mechanism of uptake or toxicity. Therefore, we need to consider relevant sublethal endpoints when assessing these CuO NPs to ensure that we do not overlook long-term effects.

Magnesium oxide nanoparticles and their ecotoxicological effect on edaphic organisms in tropical soil.

The demand for food has intensified production in agricultural areas and stimulated the use of nanotechnology to develop new inputs, especially nanoparticle materials. In this new context, predicting the impact of using nanoparticles on non-target organisms becomes a necessary measure. The aim of this study was to evaluate the ecotoxicological potential of magnesium (Mg2+ ) added via magnesium oxide nanoparticles (MgO-NPs), magnesium oxide (MgO), and magnesium nitrate hexahydrate (Mg [NO3 ]2 ·6H2 O) incubated over time in tropical soil on earthworms (Eisenia andrei), springtails (Folsomia candida), and enchytraeids (Enchytraeus crypticus). Tests were conducted using a clay-textured Latossolo Vermelho distrófico (Oxisol), which received increasing doses of Mg2+ (0; 25; 50; 100; 200 and 400 mg kg−1 of soil) from the three sources tested added to the soil. Treated soil was incubated for 120 days in a room with controlled temperature and photoperiod, and the ecotoxicological tests were performed at 0, 60, and 120 days of incubation. Despite having caused reduction in the reproduction of F. candida at the incubation time 0, MgO-NPs showed a low toxic potential against the other species studied, with toxicity only at a higher dose of 50 mg Mg kg−1 when compared to the other sources of Mg2+ applied to the soil (MgO and Mg [NO3 ]2 ·6H2 O). Responses associated with incubation times showed that all magnesium sources tested have lower toxicity over incubation time.

Acute toxicity of the insecticide abamectin and the fungicide difenoconazole (individually and in mixture) to the tropical stingless bee Melipona scutellaris.

Stingless bees have been recognized as essential plant pollinators and producers of various natural products in neotropical areas. Research into the potential risks of pesticides they may be exposed to in agricultural fields, however, remains meagre. Especially the toxicity of pesticide mixtures likely to occur under real-world conditions and that are likely to exert synergetic effects has been poorly studied. The aim of the present study was therefore to evaluate the single and mixture acute contact and oral toxicity of commercial products containing the insecticide abamectin and the fungicide difenoconazole in laboratory bioassays with the Brazilian native stingless bee Melipona scutellaris. In addition, a comparison of the insecticide sensitivity of stingless bees relative to the honeybee Apis mellifera was made based on previously published toxicity data. Except for oral exposure to abamectin, M. scutellaris appeared to be more sensitive that A. mellifera in the single compound toxicity tests. A difenoconazole concentration at the NOEC (no observed effect concentration) level indicated a synergetic toxic interaction with abamectin. A sensitivity comparison based on published toxicity data for A. mellifera and stingless bees indicated several insecticidal modes of action having a high relative sensitivity to stingless bees that need especial consideration in future studies. The research findings highlight the need for testing native bee species and environmentally relevant pesticide mixtures in risk assessments to avoid underestimation of potential risks to bee populations and the subsequent loss of pollination ecosystem services.

Changes in soil mesofauna structure due to different land use systems in south Minas Gerais, Brazil.

Essential to the provision of important ecosystems services, i.e., food production, soil is suffering great pressure. The degradation of natural areas in order to turn them into croplands has been causing severe effects to the soil quality, including the maintenance of their biodiversity. Soil physical disruption reduce the soil biodiversity and, consequently, may cause negative effects to the supporting services, i.e., organic matter degradation and nutrient cycling, which will directly or indirectly impact agroecosystems. In this study, the influence of three different types of land uses (native forest (NF), conventional agriculture (CA), and organic agriculture (OA)) to the soil mesofauna (emphasizing collembolan and mites) were assessed under real scenarios in the southeast part of Brazil. Both conventional and organic fields were of strawberries, and the greatest difference in their processes was the use of synthetic fertilizers and pesticides, performed at CA. Soil fauna organisms were collected and identified to main groups, except the collembolan species which were further separated into four main groups/family. Results showed that not only the physical changes due to soil tillage caused negative effects to soil fauna. In the field where the use of agricultural products was allowed, organisms were much more severely affected. Hence, the conversion of natural forested areas to agricultural lands may harm soil fauna communities through biodiversity loss. This study not only adds significant information to the knowledge regarding the relation between biodiversity loss and agricultural practices worldwide, but it also helps to improve Brazilian knowledge of the edaphic fauna in agroecosystems.

Dermal Fungicide Exposure at Realistic Field Rates Induces Lethal and Sublethal Effects on Juvenile European Common Frogs (Rana temporaria).

Viticulture is one of the most pesticide-intensive agricultures in Europe, leading to a spatiotemporal overlap of amphibian migration and pesticide applications. Because postmetamorphic, terrestrial amphibian stages are mostly neglected in ecotoxicological studies, we investigated acute effects of viticultural fungicides on juvenile common frogs (Rana temporaria). Tadpoles from an uncontaminated pond were placed in enclosures in 8 ponds with an increasing degree of pesticide contamination in southwest Germany to represent different aquatic exposure backgrounds. After metamorphosis, juveniles were exposed to soil contaminated with 50% of the recommended field rates of the fungicides Folpan® 80 water dispersible granule (WDG) and Folpan® 500 suspension concentrate with the same amount of folpet as active ingredient and differing additives. After 48 h, effects on the survival, body mass, and behavior were investigated. No effect of the aquatic exposure background on terrestrial sensitivity could be detected. Acute terrestrial exposure led to mean mortality rates of 14% (13-17%, suspension concentrate) and 60% (17-100%, WDG) and resulted in adverse effects on locomotor activity as well as feeding behavior. Moreover, the results suggest that the toxicity of the 2 tested folpet formulations depends on their additives. Because the identified effects may result in severe impairments and thus in declines of amphibian populations, a more protective risk assessment of pesticides is needed for postmetamorphic amphibians to ensure proper conservation of amphibian populations. Environ Toxicol Chem 2021;40:1289-1297. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Different diets with and without inclusion of antimicrobial additives alter the toxicity of swine manure to springtails and earthworms? / Diferentes dietas com e sem inclusão de aditivos antimicrobianos alteram a toxicidade do esterco de suíno para colêmbolos e minhocas?

The objective of this study was to measure the impact on part of soil fauna of application of non-stabilized (fresh) manure from post-weaning pigs fed diets, formulated with or without the use of dual-purpose wheat (15% inclusion), and with or without the use of antimicrobial growth-promoting additives (100 mg kg-1 doxycycline + 50 mg kg-1 colistin + 250 mg kg-1 Zn oxide). Two species of edaphic organisms were evaluated, the springtails Folsomia candida and the earthworms Eisenia andrei, using ecotoxicological avoidance behavior tests. The treatments were swine manure from: RR: Reference Ration; WR: Wheat Reference; RA: Reference Ration + Antimicrobial Additives; WA: Wheat Reference + Antimicrobial Additives. The doses of waste used for treatments were as follows: 0 (control), 5, 10, 20, 30, 40, 65 and 100 m³ ha-1 for the springtails, and 0 (control), 5, 10, 20, 30, 40 and 65 m³ ha-1 for the earthworms, applied in the Oxisol. The experimental design was completely randomized with four replicates. The use of non-stabilized swine manure did not affect the avoidance behavior of F. candida at any dose, regardless of the use of antimicrobial additives or wheat. For E. andrei, there was avoidance behavior at all treatments and doses used. These avoidance behaviors were related to the sensitivity of each species of soil organism. The avoidance behavior for earthworms was related to the doses of non-stabilized swine manure in soil and not to the various diets and/or the use of growth-promoting additives

O estudo foi realizado com objetivo de mensurar o impacto sobre parte da fauna do solo de aplicações de dejetos sem estabilização (frescos), provenientes de suínos na fase de creche arraçoados com diferentes dietas, formuladas com ou sem o uso do trigo de duplo propósito (15% de inclusão), e com ou sem o uso de aditivos antimicrobianos promotores de crescimento (100 mg kg-1 de doxiciclina + 50 mg kg-1de colistina + 250 mg kg-1 de óxido de Zn). Foram avaliadas duas espécies de organismos edáficos, colêmbolos Folsomia candida e minhocas Eisenia andrei, com o uso de ensaios ecotoxicológicos de comportamento de fuga. Os tratamentos foram dejetos de suínos provenientes de: RR: Ração Referência; WR: Trigo Referência; RA: Ração Referência + Aditivos Antimicrobianos; WA: Trigo Referência + Aditivos Antimicrobianos. As doses de dejetos utilizadas para os tratamentos foram: 0 (controle), 5, 10, 20, 30, 40, 65 e 100 m³ ha-1 para os colêmbolos, e 0 (controle), 5, 10, 20, 30, 40 e 65 m³ ha-1 para as minhocas, aplicadas em Latossolo Vermelho distrófico. O delineamento experimental foi inteiramente casualizado com quatro réplicas. A utilização de dejetos de suínos não estabilizados não afetou a fuga de F. candida em nenhuma das doses, independente do uso ou não de aditivos antimicrobianos ou de trigo. Já para E. andrei o comportamento foi inverso e houve fuga em todos os tratamentos e doses utilizados. Os comportamentos de fuga foram relacionados à sensibilidade de cada espécie de organismo edáfico. O comportamento de fuga para minhocas foi relacionado com as doses dos dejetos de suínos não estabilizados no solo e não às várias dietas e/ou ao uso dos aditivos promotores de crescimento

Impacts on reproduction of Enchytraeus crypticus in fertilized soils with chicken litter treated with synthetic and natural insecticide.

Poultry litter is used as a fertilizer due to the high concentration of nutrients. However, this material receives application of medicines, whether they are used to treat animals or the litter itself. Thus, the objective was to evaluate if the application in soils of poultry litter doses treated with two insecticides (cypermethrin and cinnamon oil) used to control of the Alphitobius diaperinus, affect the reproduction of Enchytraeus crypticus, and the insecticides effects over time, trough of ecotoxicological tests using soils with different textural classes an Entisol and an Oxisol. In the Oxisol the application of poultry litter stimulated the E. crypticus reproduction regardless of the insecticide and the collection time. In Entisol the poultry litter showed a toxicity at 10 t ha-1 when treated with cypermethrin and at 30 t ha-1 with cinnamon oil. Entisol results show that cypermethrin affects non-target edaphic organisms soon after spraying, but over time has reduced toxicity unlike cinnamon oil.

Ecotoxicity test as an aid in the determination of copper guideline values in soils / Ensaios de ecotoxicidade como auxílio na determinação de valores orientadores de cobre em solos

ABSTRACT: The process of identifying a contaminated area involves a preliminary assessment consisting of chemical analysis and comparison with guideline values. In Brazil, these values are included in the CONAMA Resolution no. 420/2009, based on chemical analysis and phytotoxicity. The objective of this research was to evaluate the ecotoxicity of copper in two natural soils in the state of Rio de Janeiro, a Haplic Planosol (sandy) and a Red-Yellow Argisol (medium texture), to verify if the values established by the legislation offer protection to the soil fauna. Lethality and reproduction tests were performed with Eisenia andrei worms and Folsomia candida springtails. Results indicated copper values in Planosol as LC50 435 mg kg 1 and EC50 29 mg kg 1 for E. Andrei and LC10 137 mg kg 1 and EC50 117 mg kg 1 for F. candida. In Argisol, these results were LC50 690 mg kg 1 and EC50 61 mg kg 1 for E. andrei, and LC10 42 mg kg 1 and EC50 138 mg kg 1 for F. candida. Values reported are lower than the research value (200 mg kg-1) established by the CONAMA resolution 420/09, indicating that concentrations lower than the limit values may affect these organisms, depending on the type of soil. Worms were more sensitive than springtails to copper contamination, and sandy soil was more susceptible to ecotoxicity due to copper contamination, probably due to the greater bioavailability of the metal. Results of such ecotoxicity tests should be considered in the development of soil guideline values.

RESUMO: O processo de identificação de uma área contaminada envolve uma avaliação preliminar que consiste em análise química e comparação com valores orientadores. No Brasil, esses valores constam na Resolução CONAMA no 420/2009, baseados em análises químicas e fitotoxicidade. O objetivo deste trabalho foi avaliar a ecotoxicidade do cobre em dois solos naturais do Estado do Rio de Janeiro, um Planossolo Háplico (arenoso) e um Argissolo Vermelho-Amarelo (textura média), para verificar se os valores estabelecidos pela legislação oferecem proteção à fauna do solo. Foram realizados ensaios de letalidade e de reprodução com minhocas da espécie Eisenia andrei e com colêmbolos da espécie Folsomia candida. Os resultados indicaram no Planossolo valores de cobre para E. andrei de CL50 435 mg kg-1 e CE50 29 mg kg-1 e para F. candida CL10 137 mg kg-1 e EC50 117 mg kg-1. No Argissolo esse resultado foi para E. andrei CL50 690 mg kg-1 e CE50 61 mg kg-1, e para F. candida CL10 42 mg kg-1 e CE50 138 mg kg-1. Os valores encontrados são menores que o valor de investigação (200 mg kg-1) estabelecidos pela resolução 420/09 do CONAMA, indicando que concentrações menores do que os valores-limite podem afetar esses organismos, a depender do tipo de solo. As minhocas mostraram-se mais sensíveis que os colêmbolos à contaminação por cobre, e o solo arenoso mostrou-se mais suscetível a apresentar ecotoxicidade diante de uma contaminação por cobre, devida, provavelmente, à maior biodisponibilidade do metal. Os ensaios de ecotoxicidade devem ser considerados no desenvolvimento de valores orientadores para solos.

Ecotoxicological effect of fipronil and its metabolites on Folsomia candida in tropical soils.

The aim of this study is to evaluate the toxicity of the fipronil and its metabolites, fipronil sulfone and fipronil desulfinyl on Folsomia candida. Three types of soils were used to perform the ecotoxicological tests, two natural soils (Oxisol and Entisol), and an artificial substrate (Tropical Artificial Soil). The treatments consisted of increasing doses of a veterinary medicinal product with active ingredient based on fipronil, and of its two main metabolites, fipronil sulfone and fipronil desulfinyl. The doses of fipronil were 0.00, 0.02, 0.04, 0.08, 0.15, 0.30, 0.60, and 1.00 mg kg-1 of the active ingredient. For fipronil sulfone and fipronil desulfinyl the doses tested were: 0.00, 0.01, 0.02, 0.04, 0.06, 0.08, 0.15, 0.30, 0.60, 1.00, and 5.00 mg kg-1. The effects of toxicity on F. candida were evaluated by survival and reproduction tests, based on the methods described by ISO 11267. There was a completely randomized design, with six replicates for the fipronil tests and eight for the metabolites. We show that fipronil and its metabolites, even at very low concentrations, are toxic to F. candida. Reduction of reproduction can be observed at concentrations of 0.15 mg kg-1. There was little difference in toxicity (LC50 and EC50) between fipronil and its metabolites. Rather, differences were related more to soil type, where the artificial soil was more sensitive than the two natural soils (Oxisol and Entisol). We conclude that it is important to perform ecotoxicological tests in natural soils, in order to generate more realistic representations of veterinary drug toxicity on the environment.

Mixture toxicity of four commonly used pesticides at different effect levels to the epigeic earthworm, Eisenia fetida.

As commonly used pesticides, chlorpyrifos (CPF), fenobucarb (FEN), clothianidin (CLO) and acetochlor (ACE) are widely applied on crops worldwide. In this study, the combined toxicities of their binary, ternary and quaternary mixtures were evaluated using the earthworm Eisenia fetida as test organism. Mixture toxicities were studied using the combination index (CI) method and visualized by isobolograms, and then data were compared with traditional concentration addition (CA) and independent action (IA) models. Two binary mixtures of CPF+FEN and FEN+ACE, two ternary mixtures of CPF+CLO+FEN and CPF+FEN+ACE, and quaternary mixture of CPF+FEN+ACE+CLO exhibited a clear synergistic effect. The CI method was compared with the classical models of CA and IA, indicating that the CI method could accurately predict the combined toxicities of the chemicals. The results indicated that it was difficult to predict combined effects of these pesticides from mode of action alone because of existence of complicated synergistic and antagonistic responses. More attention should be paid to the potential synergistic effects of chemicals interactions, which might cause serious ecological problems.

Ecotoxicology of mercury in tropical forest soils: Impact on earthworms.

Mercury (Hg) is one of the most toxic nonessential trace metals in the environment, with high persistence and bioaccumulation potential, and hence of serious concern to environmental quality and public health. Emitted to the atmosphere, this element can travel long distances, far from emission sources. Hg speciation can lead to Hg contamination of different ecosystem components, as well as biomagnification in trophic food webs. To evaluate the effects of atmospheric Hg deposition in tropical forests, we investigated Hg concentrations in earthworm tissues and soils of two Forest Conservation Units in State of Rio de Janeiro, Brazil. Next, we performed a laboratory study of the biological responses (cast analysis and behavioral, acute, chronic and bioaccumulation ecotoxicological tests) of two earthworms species (Pontoscolex corethrurus and Eisenia andrei) to Hg contamination in tropical artificial soil (TAS) and two natural forest soils (NS) spiked with increasing concentration of HgCl2. Field results showed Hg concentrations up to 13 times higher in earthworm tissues than in forest soils, while in the laboratory Hg accumulation after 91-days of exposure was 25 times greater in spiked-soils with 128mgHgkg-1 (dry wt) than in control (unspiked) soils. In all the toxicity tests P. corethrurus showed a higher adaptability or resistance to mercury than E. andrei. The role of earthworms as environmental bioremediators was confirmed in this study, showing their ability to greatly bioaccumulate trace metals while reducing Hg availability in feces.

Combined toxicity of imidacloprid and three insecticides to the earthworm, Eisenia fetida (Annelida, Oligochaeta).

Although the earthworm Eisenia fetida has been used in many ecotoxicological studies in recent years, most of these studies have only focused on assessing the effects of individual insecticides. In the present study, we aimed to compare the individual and combined toxic effects of imidacloprid and three insecticides (phoxim, chlorpyrifos, and lambda-cyhalothrin) on E. fetida. We showed that imidacloprid had the highest intrinsic toxicity to the worms in filter paper contact test, followed by phoxim and lambda-cyhalothrin, while the least toxicity was found from chlorpyrifos. Moreover, 14-day soil toxicity test revealed that the highest toxicity was still detected for imidacloprid with an LC50 value of 2.82 (2.61∼3.17) mg a.i. kg-1 dry weight (DW), followed by chlorpyrifos with an LC50 value of 384.9 (353.5∼440.3) mg a.i. kg-1 DW. Meanwhile, a relatively less toxicity was found for lambda-cyhalothrin with an LC50 value of 560.3 (475.9∼718.5) mg a.i. kg-1 DW, while the lowest toxicity to E. fetida was observed for phoxim with an LC50 value of 901.5 (821.3∼1017) mg a.i. kg-1 DW. In addition, significant synergistic responses were found from the ternary mixture of imidacloprid-phoxim-lambda-cyhalothrin and quaternary mixture of imidacloprid-phoxim-chlorpyrifos-lambda-cyhalothrin in both bioassay systems. Therefore, our findings highlighted that the simultaneous presence of several insecticides in the soil environment might lead to increased toxicity, resulting in serious damage to the nontarget organisms compared with individual insecticides.

Lethal and sub-lethal effects of five pesticides used in rice farming on the earthworm Eisenia fetida.

The toxicity of five pesticides typically used in rice farming (trichlorfon, dimethoate, carbendazim, tebuconazole and prochloraz) was evaluated on different lethal and sub-lethal endpoints of the earthworm Eisenia fetida. The evaluated endpoints included: avoidance behaviour after an exposure period of 2 days; and mortality, weight loss, enzymatic activities (cholinesterase, lactate dehydrogenase and alkaline phosphatase) and histopathological effects after an exposure period of 14 days. Carbendazim was found to be highly toxic to E. fetida (LC50=2mg/kg d.w.), significantly reducing earthworm weight and showing an avoidance response at soil concentrations that are close to those predicted in rice-fields and in surrounding ecosystems. The insecticide dimethoate showed a moderate acute toxicity (LC50=28mg/kg d.w.), whereas the rest of tested pesticides showed low toxicity potential (LC50 values above 100mg/kg d.w.). For these pesticides, however, weight loss was identified as a sensitive endpoint, with NOEC values approximately 2 times or lower than the calculated LC10 values. The investigated effects on the enzymatic activities of E. fetida and the observed histopathological alterations (longitudinal and circular muscle lesions, edematous tissues, endothelial degeneration and necrosis) proved to be sensitive biomarkers to monitor pesticide contamination and are proposed as alternative measures to evaluate pesticide risks on agro-ecosystems.

The use of diplopods in soil ecotoxicology - a review.

Diplopods play an important role in the dynamics of terrestrial ecosystems, actively participating in the decomposition of organic matter and soil aeration. They have gained increased attention from ecotoxicology research because they are continuously exposed to soil contaminants and biological effects of chemical stressors can be measurable at various levels of biological organization. This paper is the first review on the use of diplopods as soil bioindicators and compiles the effects of the different toxic chemical agents on these animals. Special emphasis is given on the interpretation of the effects of heavy metals and complex mixtures in target organs of diplopods.