Effect of Sargassum on the Behavior and Survival of the Earthworm Eisenia Fetida.

The massive influx of Sargassum natans and S. fluitans to the shores of the Mexican Caribbean has raised concerns regarding their potential impact on soil quality and health in coastal and agroecosystems. The effects of Sargassum accumulation remain largely unexplored. This study aimed to assess the impact of Sargassum on soil ecosystems by examining the behavior and survival of the epigean earthworm Eisenia fetida. The earthworm was exposed to varying concentrations of Sargassum (0, 25, 50, 75, and 100%) in two toxicological tests. Results from the avoidance test demonstrated that E. fetida exhibited strong aversion (> 80%) to a diet containing 100% Sargassum. Conversely, the acute test revealed minimal mortality, but growth decreased with increasing Sargassum concentrations. These findings can serve as early warning bioindicators for assessing the environmental risk posed by Sargassum in soil ecosystems.

Genomes were 'scrambled' when worms left the sea.

Chromosomal chaos may have aided their moves to fresh water and land.

Rhizosphere microbiomes derived from vermicompost alter gene expression and regulatory pathways in tomato (Solanum lycopersicum, L.).

The gut microbiome of worms from composting facilities potentially harbors organisms that are beneficial to plant growth and development. In this experiment, we sought to examine the potential impacts of rhizosphere microbiomes derived from Eisenia fetida worm castings (i.e. vermicompost) on tomato (Solanum lycopersicum, L.) plant growth and physiology. Our experiment consisted of a greenhouse trial lasting 17 weeks total in which tomato plants were grown with one of three inoculant treatments: a microbial inoculant created from vermicompost (V), a microbial inoculant created from sterilized vermicompost (SV), and a no-compost control inoculant (C). We hypothesized that living microbiomes from the vermicompost inoculant treatment would enhance host plant growth and gene expression profiles compared to plants grown in sterile and control treatments. Our data showed that bacterial community composition was significantly altered in tomato rhizospheres, but fungal community composition was highly variable in each treatment. Plant phenotypes that were significantly enhanced in the vermicompost and sterile vermicompost treatments, compared to the control, included aboveground biomass and foliar δ15N nitrogen. RNA sequencing revealed distinct gene expression changes in the vermicompost treatment, including upregulation of nutrient transporter genes such as Solyc06g074995 (high affinity nitrate transporter), which exhibited a 250.2-fold increase in expression in the vermicompost treatment compared to both the sterile vermicompost and control treatments. The plant transcriptome data suggest that rhizosphere microbiomes derived from vermicompost can influence tomato gene expression and growth-related regulatory pathways, which highlights the value of RNA sequencing in uncovering molecular responses in plant microbiome studies.

Micro/nanoplastics pollution poses a potential threat to soil health.

Micro/nanoplastic (MNP) pollution in soil ecosystems has become a growing environmental concern globally. However, the comprehensive impacts of MNPs on soil health have not yet been explored. We conducted a hierarchical meta-analysis of over 5000 observations from 228 articles to assess the broad impacts of MNPs on soil health parameters (represented by 20 indicators relevant to crop growth, animal health, greenhouse gas emissions, microbial diversity, and pollutant transfer) and whether the impacts depended on MNP properties. We found that MNP exposure significantly inhibited crop biomass and germination, and reduced earthworm growth and survival rate. Under MNP exposure, the emissions of soil greenhouse gases (CO2, N2O, and CH4) were significantly increased. MNP exposure caused a decrease in soil bacteria diversity. Importantly, the magnitude of impact of the soil-based parameters was dependent on MNP dose and size; however, there is no significant difference in MNP type (biodegradable and conventional MNPs). Moreover, MNPs significantly reduced As uptake by plants, but promoted plant Cd accumulation. Using an analytical hierarchy process, we quantified the negative impacts of MNP exposure on soil health as a mean value of -10.2% (-17.5% to -2.57%). Overall, this analysis provides new insights for assessing potential risks of MNP pollution to soil ecosystem functions.

SoxC and MmpReg promote blastema formation in whole-body regeneration of fragmenting potworms Enchytraeus japonensis.

Regeneration in many animals involves the formation of a blastema, which differentiates and organizes into the appropriate missing body parts. Although the mechanisms underlying blastema formation are often fundamental to regeneration biology, information on the cellular and molecular basis of blastema formation remains limited. Here, we focus on a fragmenting potworm (Enchytraeus japonensis), which can regenerate its whole body from small fragments. We find soxC and mmpReg as upregulated genes in the blastema. RNAi of soxC and mmpReg reduce the number of blastema cells, indicating that soxC and mmpReg promote blastema formation. Expression analyses show that soxC-expressing cells appear to gradually accumulate in blastema and constitute a large part of the blastema. Additionally, similar expression dynamics of SoxC orthologue genes in frog (Xenopus laevis) are found in the regeneration blastema of tadpole tail. Our findings provide insights into the cellular and molecular mechanisms underlying blastema formation across species.

A comparative study of worm-sludge treatment reed bed planted with Phragmites australis and Arundo donax in the Mediterranean region.

Sludge treatment reed bed planted (STRB) with Phragmites australis (P.australis) and Arundo donax (A.donax) was assessed in the presence of Eisenia fetida under control condition during the dry season. Worm-planted units were fed with mixed sewage sludge (dry and volatile solids of 29.44 g DS.L-1 and 24.23 g VS.L-1). Sludge loading rates (SLR) of 50, 60, and 70 kg DS m-2 year-1 were examined to assess dewatering efficiency. Surface layers in units with P.australis and A.donax achieved DS of 80 and 81% at a loading rate of 50 kg DS m-2 year-1, while their subsurface DS were 41 and 25%, respectively. Units with A.donax experienced plant loss when subjected to SLR exceeding 60 kg DS m-2 year-1. More than 10 cm of residual sludge accumulated on the top of units after a 2-month final rest. Evapotranspiration was greater in the unit with P.australis (5.23 mm day-1) compared to the unit with A.donax (4.24 mm day-1) while both were fed with 70 kg DS m-2 year-1. Water loss contributions from residual sludge layer, drained water, and evapotranspiration were 3, 46, and 51%, respectively. Units with P.australis indicated 20% higher water loss compared to units with A.donax. Although the drained water quality improved over time, it did not meet standard limits. The residual sludge layer contained macro and micronutrients, and heavy metals with a significant elemental order of N > Ca > P > S > mg > K (N:P:K = 31:8:1), Fe > Na > B > Mn > Mo and Zn > Cr > Cu > Pb > Ni > Cd. Overall, STRB could be a sustainable alternative technology to conventional sewage sludge management techniques.

Analysis of microplastics in the reuse of compost in three agricultural sites (Cádiz, Spain) as a circular economy strategy: detection of micropollutants and incidence of plastic ingestion levels by annelids.

The system of fertilizing agricultural soils with sludge or compost from wastewater treatment processes, as one of the principles of the circular economy, can lead to microplastic (MP) contamination. The existing technical standards for fertilization are very recent and do not consider this problem, although there is scientific evidence of their existence. Therefore, this study, on the one hand, evaluates the presence of MPs in agricultural soils, previously treated with sludge or compost from wastewater treatment plants for fertilization, and on the second hand, it studies the effect of these MPs on earthworms in three different locations in the south of Spain. For the study, selected composts deriving from the different stages of the composting process and three fertilized soils with increasing MP doses were followed. Samples were taken from different sections in depth (0-5, 5-10, and 10-20 cm) to study the shape, size, type, and abundance of MPs using infrared spectroscopy (FTIR). The results showed that the most abundant shape was fiber, followed by fragment and finally bulk, for both composts and soils. Regarding size distribution, 100 µm was the predominant size in composts (64.3% ± 9.8), while in the case of soils, the predominant range was from 100 to 500 µm. The prevalent polymers in both, composts and soils, were PTFE, TPE, PP, and PET, with four times higher amounts in composts than in soils. Ingestion of common MPs were also verified in two earthworm species, which ingested concentrations higher than 2.1% w/w. PP was the most ingested MP and Eisenia fetida was more voracious compared with Lumbricus terrestris. Therefore, it can be considered a suitable bioindicator for monitoring microplastic contamination in agricultural soil.

Vermifiltration as a green solution to promote digestate reuse in agriculture in small-scale farms.

Digestates from low-tech digesters need to be post-treated to ensure their safe agricultural reuse. This study evaluated, for the first time, vermifiltration as a post-treatment for the digestate from a low-tech digester implemented in a small-scale farm, treating cattle manure and cheese whey under psychrophilic conditions. Vermifiltration performance was monitored in terms of solids, organic matter, nutrients, and pathogens removal efficiency. In addition, the growth of earthworms (Eisenia foetida) and their role in the process was evaluated. Finally, the vermicompost and the effluent of the vermifilter were characterized in order to assess their potential reuse in agriculture. Vermifilters showed high removal efficiency of chemical oxygen demand (55-90%), total solids (60-80%), ammonium nitrogen (83-97%), and phosphate-P (28-49%). Earthworms effectively grew and reproduced on digestate (i.e. earthworms number increased by 183%), enhancing the vermifiltration performance, while reducing clogging and odour-related issues. Both the vermicompost and effluent produced complied with legislation limits established for soil improvers and wastewater for fertigation, respectively. Indeed, there was an absence of pathogens and non-detectable heavy metals concentrations. Vermifiltration may be thus considered a suitable post-treatment option for the digestate from low-tech digesters, allowing for its safe agricultural reuse and boosting the circular bioeconomy in small-scale farms.

Vermistabilization of excess sludge employing Eisenia fetida: Earthworm histopathological alterations and phytotoxicity evaluation.

The aim of this work was to stabilize excess sludge (ES) coming from a wastewater treatment plant (WWTP) by vermistabilization and to evaluate ecotoxicological effects over the earthworm species Eisenia fetida. Three mixtures were made up in triplicate using different volume ratios of ES and soil (S) (100% ES, 70:30% ES:S and 30:70% ES:S in wet weight basis). Earthworms were added in order to compare vermicomposting vs. natural stabilization. The mixtures were monitored over 130 days through physical, chemical, pathological and biological analysis, following quality standards depicted in the US EPA 40 CFR Part 503, local regulations and background studies. Histopathological samples were processed as biomarkers of acute and chronic toxicity on earthworms, and germination assays were performed at the end of the experiment to assess phytotoxicity. In terms of pathogen depletion comparing initial and final values from each treatment, the mixtures with higher ES proportions (70 and 100%) with earthworms were the most efficient ones registering 64.8 and 75.5% of reduction of fecal coliforms (FC) respectively, while the lowest ES proportion with earthworms (30%) showed 54.7%. Final pathogens content in all the treatments with earthworms were significantly lower (ranged from 1360 to 1760 MPN g total solids-1) than the values registered in treatments without earthworms (ranged from 2400 to 4000 MPN g total solids-1) (p < 0.05). However, none of the treatments attained class A categorization (FC ≤ 1000 MPN g total solids-1) in terms of FC. Also, values of mean cocoon production and hatched juveniles along time were significantly higher in the treatments with 100 and 70% ES (p < 0.05), while the higher mean adult biomass was detected in the treatment with 30% ES. Volatile solids decrease ranged between 8.45 and 22.34% in treatments with earthworms and all values of specific oxygen uptake rate were below 1.5 mg O2 h -1 g total solids -1. There were not negative effects over behavior or reproduction of E. fetida adults, nor the presence of external and internal injuries. Final products from mixtures with earthworms presented a humus-like structure, were odorless and reached maturity values -presenting no phytotoxicity-with significant differences between germination index values of treatments with and without earthworms (p < 0.05). Vermistabilization is a successful eco-technology to sanitize excess sludge, acquiring an added-value material and contributing to its revalorization as organic amendments or fertilizers in soils within the circular economy framework and the United Nations' Sustainability Development Goals.

The interaction between biochar and earthworms: Revealing the potential ecological risks of biochar application and the feasibility of their co-application.

Biochar's interaction with soil-dwelling organisms, particularly earthworms, is crucial in ensuring the effective and secure utilization of biochar in the soil. This review introduces the application of biochar in soil, summarizes how earthworms respond to biochar-amended soil and the underlying factors that can influence their response, discusses the synergistic and antagonistic impacts of earthworm activity on the efficacy of biochar, and considers the feasibility of applying them together. A review of existing research has identified uncertainty in the effect of biochar exposure on earthworms, with biochar derived from animal wastes, produced at higher pyrolysis temperatures, and used at higher doses of biochar having more negative effects on earthworms. Habitat modification, toxicity release, particle effects, and contaminant immobilization are underlying factors in how biochar affects earthworm indicators. While biochar in contaminated soils may alleviate the stress of pollutants on earthworms by decreasing their bioaccumulation, this remedial effect is not always effective. Additionally, earthworm bioturbation can enhance the migration, fragmentation, and oxidation of biochar, while also stimulating extracellular enzymes that convert biochar into 'vermichar'. Earthworms and biochar can synergize well to improve soil fertility and remediate soil organic pollution, yet exhibit contrasting roles in soil C sequestration and immobilizing heavy metals in soil. These findings highlight both the advantages and risks of their co-application. Therefore, when considering the use of biochar alone or with earthworms, it is crucial to thoroughly assess its potential ecotoxicity on earthworms and other soil organisms, as well as the influence of bioturbation, such as that caused by earthworms, on the effectiveness of biochar.

Response and defense mechanisms of the earthworms Eisenia foetida to natural saline soil stress.

Salinization of soil is a serious global environmental issue, particularly in agricultural lands. Saline farmland not only endangers grain production but also affects the survival of soil fauna. Earthworms, as soil ecosystem engineers, play a crucial role in maintaining soil health and enhancing global agricultural production. However, the response of earthworms to natural saline soil stress remains poorly understood. To explore this, we investigated the effects of natural saline soil from Dongying City, Shandong Province, China, on the growth, survival, reproduction, antioxidation, and defense-related gene expression of the earthworm Eisenia foetida. Our findings demonstrate that the growth rate, survival rate, and cocoon production of E. foetida decrease under exposure to natural saline soil in a dose-dependent manner. Elevated levels of DNA damage in coelomocytes and increased reactive oxygen species (ROS) were observed. Additionally, antioxidant enzymes, such as superoxide dismutase (SOD) and catalase (CAT), increased under stress. The mRNA levels of Cyp450 and Hsp70 also rose in response to saline soil exposure. Furthermore, the activity of Na+/K+-ATPase and the expression of the osmotic sensor gene wnk-1 were elevated. In conclusion, our findings indicate that natural saline soil induces antioxidant and osmotic stress in earthworms E. foetida, highlighting the detrimental effects and defense mechanisms of soil fauna under such conditions.

Integrated assessment of the chemical, microbiological and ecotoxicological effects of a bio-packaging end-of-life in compost.

The present study aimed to i) assess the disintegration of a novel bio-packaging during aerobic composting (2 and 6 % tested concentrations) and evaluate the resulting compost ii) analyse the ecotoxicity of bioplastics residues on earthworms; iii) study the microbial communities during composting and in 'earthworms' gut after their exposure to bioplastic residues; iv) correlate gut microbiota with ecotoxicity analyses; v) evaluate the chemico-physical characterisation of bio-packaging after composting and earthworms' exposure. Both tested concentrations showed disintegration of bio-packaging close to 90 % from the first sampling time, and compost chemical analyses identified its maturity and stability at the end of the process. Ecotoxicological assessments were then conducted on Eisenia fetida regarding fertility, growth, genotoxic damage, and impacts on the gut microbiome. The bioplastic residues did not influence the earthworms' fertility, but DNA damages were measured at the highest bioplastic dose tested. Furthermore bioplastic residues did not significantly affect the bacterial community during composting, but compost treated with 2 % bio-packaging exhibited greater variability in the fungal communities, including Mortierella, Mucor, and Alternaria genera, which can use bioplastics as a carbon source. Moreover, bioplastic residues influenced gut bacterial communities, with Paenibacillus, Bacillus, Rhizobium, Legionella, and Saccharimonadales genera being particularly abundant at 2 % bioplastic concentration. Higher concentrations affected microbial composition by favouring different genera such as Pseudomonas, Ureibacillus, and Streptococcus. For fungal communities, Pestalotiopsis sp. was found predominantly in earthworms exposed to 2 % bioplastic residues and is potentially linked to its role as a microplastics degrader. After composting, Attenuated Total Reflection analysis on bioplastic residues displayed evidence of ageing with the formation of hydroxyl groups and amidic groups after earthworm exposure.

Ecological risk assessment of castor oil based waterborne polyurethane: Mechanism of anionic/cationic state selective toxicity to Eisenia fetida.

Cationic and anionic castor oil-based waterborne polyurethanes (C-WPU/A and C-WPU/C) have great potential for development in agriculture. However, it is still unclear whether these polyurethanes are harmful or toxic to soil fauna. Based on multilevel toxicity endpoints and transcriptomics, we investigated the effects of C-WPU/A and C-WPU/C on earthworms (Eisenia fetida). The acute toxicity results showed that C-WPU/A was highly toxic to the earthworms, whereas C-WPU/C was nearly nontoxic. C-WPU/A significantly affected the body weight, burrowing ability and cocoon production rate of earthworms compared to C-WPU/C. After exposure to C-WPU/A, the results showed accumulation of reactive oxygen species (ROS), abnormal peroxidase activity, and increased malondialdehyde levels. Additionally, more serious histopathological damage was observed in earthworms, such as epidermal damage, vacuolization, longitudinal muscle disorganization, and shedding of intestinal epidermal cells. At the cellular level, C-WPU/A induced more severe lysosomal damage, DNA damage and apoptosis than C-WPU/A. C-WPU/A made more differentially expressed genes and considerably more enriched pathways at the transcriptional level than C-WPU/C. These pathways are largely involved in cell membrane signaling, detoxification, and apoptosis. These results provide an important reference for elucidating the selective toxicity mechanisms of C-WPU/A and C-WPU/C in earthworms.

Machine Learning Allowed Interpreting Toxicity of a Fe-Doped CuO NM Library Large Data Set─An Environmental In Vivo Case Study.

The wide variation of nanomaterial (NM) characters (size, shape, and properties) and the related impacts on living organisms make it virtually impossible to assess their safety; the need for modeling has been urged for long. We here investigate the custom-designed 1-10% Fe-doped CuO NM library. Effects were assessed using the soil ecotoxicology model Enchytraeus crypticus (Oligochaeta) in the standard 21 days plus its extension (49 days). Results showed that 10%Fe-CuO was the most toxic (21 days reproduction EC50 = 650 mg NM/kg soil) and Fe3O4 NM was the least toxic (no effects up to 3200 mg NM/kg soil). All other NMs caused similar effects to E. crypticus (21 days reproduction EC50 ranging from 875 to 1923 mg NM/kg soil, with overlapping confidence intervals). Aiming to identify the key NM characteristics responsible for the toxicity, machine learning (ML) modeling was used to analyze the large data set [9 NMs, 68 descriptors, 6 concentrations, 2 exposure times (21 and 49 days), 2 endpoints (survival and reproduction)]. ML allowed us to separate experimental related parameters (e.g., zeta potential) from particle-specific descriptors (e.g., force vectors) for the best identification of important descriptors. We observed that concentration-dependent descriptors (environmental parameters, e.g., zeta potential) were the most important under standard test duration (21 day) but not for longer exposure (closer representation of real-world conditions). In the longer exposure (49 days), the particle-specific descriptors were more important than the concentration-dependent parameters. The longer-term exposure showed that the steepness of the concentration-response decreased with an increased Fe content in the NMs. Longer-term exposure should be a requirement in the hazard assessment of NMs in addition to the standard in OECD guidelines for chemicals. The progress toward ML analysis is desirable given its need for such large data sets and significant power to link NM descriptors to effects in animals. This is beyond the current univariate and concentration-response modeling analysis.

Combined transcriptome and metabolome analysis reveals the toxic effects of antimony on the earthworm.

Antimony (Sb) poses a significant ecological threat. This study combines biochemical, pathological, transcriptome, and metabolome analyses to assess the short-term (14-day) toxic impact of two Sb levels (25 mg/kg and 125 mg/kg) on earthworms (Eisenia fetida). Higher Sb concentration caused severe intestinal damage, elevated metallothionein (MT) levels, and reduced antioxidant capacity. Metabolome analysis identifies 404 and 1698 significantly differential metabolites in the two groups. Metabolites such as S(-)-cathinone, N-phenyl-1-naphthylamine, serotonin, 4-hydroxymandelonitrile, and 5-fluoropentylindole contributed to the metabolic responses to Sb stress. Transcriptome analysis shows increased chitin synthesis as a protective response, impacting amino sugar and nucleotide sugar metabolism for cell wall synthesis and damage repair. Integrated analysis indicated that 5 metabolite-gene pairs were found in two Sb levels and 11 enriched pathways were related to signal transduction, carbohydrate metabolism, immune system, amino acid metabolism, digestive system, and nervous system. Therefore, the integration of multiomics approaches enhanced our comprehension of the molecular mechanisms underlying the toxicity of Sb in E. fetida.

Effects of polyvinyl chloride and low-density polyethylene microplastics on oxidative stress and mitochondria function of earthworm (Eisenia fetida).

Plastics are widely used worldwide due to their convenience. However, microplastics (MPs) accumulation poses a serious threat to ecosystem health. Therefore, understanding the effects of MPs on living organisms within their native ecosystem is crucial. Previous studies have primarily focused on the impacts of MPs in aquatic environments, whereas the effects of MPs on terrestrial ecosystems have remained largely understudied. Therefore, our study assessed the impacts of MPs on soil ecosystems by characterizing their toxic effects on earthworms (Eisenia fetida). Here, we exposed earthworms to two representative plastics within soil environments: polyvinyl chloride (PVC) and low-density polyethylene (LDPE). Given the known link between MPs and oxidative stress, we next quantified oxidative stress markers and mitochondrial function to assess the effects of MPs on the redox metabolism of earthworms. Mitochondria are crucial metabolic organelles that generate reactive oxygen species via uncontrolled ATP production. Our findings demonstrated that MPs exert different effects depending on their type. Neither the PVC-exposed groups nor the LDPE-exposed groups exhibited changes in oxidative stress, as worked by the action of superoxide dismutase (SOD) and glutathione (GSH). While treatment of the two types of MP did not significantly affect the amount of reactive oxygen species/reactive nitrogen species (ROS/RNS) generated, PVC exhibited a more pronounced effect on antioxidant system compared to LDPE. However, mitochondrial function was markedly decreased in the group exposed to high LDPE concentrations, suggesting that the examined LDPE concentrations were too low to activate compensatory mechanisms. Collectively, our findings demonstrated that exposure of MPs not only influences the antioxidant defense mechanisms of earthworms but also alters their mitochondrial function depending on their types.

Multifunctional biomolecular corona-inspired nanoremediation of antibiotic residues.

Facing complex and variable emerging antibiotic pollutants, the traditional development of functional materials is a "trial-and-error" process based on physicochemical principles, where laborious steps and long timescales make it difficult to accelerate technical breakthroughs. Notably, natural biomolecular coronas derived from highly tolerant organisms under significant contamination scenarios can be used in conjunction with nanotechnology to tackling emerging contaminants of concern. Here, super worms (Tubifex tubifex) with high pollutant tolerance were integrated with nano-zero valent iron (nZVI) to effectively reduce the content of 17 antibiotics in wastewater within 7 d. Inspired by the synergistic remediation, nZVI-augmented worms were constructed as biological nanocomposites. Neither nZVI (0.3 to 3 g/L) nor worms (104 to 105 per liter) alone efficiently degraded florfenicol (FF, as a representative antibiotic), while their composite removed 87% of FF (3 µmol/L). Under antibiotic exposure, biomolecules secreted by worms formed a corona on and modified the nZVI particle surface, enabling the nano-bio interface greater functionality, including responsiveness, enrichment, and reduction. Mechanistically, FF exposure activated glucose-alanine cycle pathways that synthesize organic acids and amines as major metabolites, which were assembled into vesicles and secreted, thereby interacting with nZVI in a biologically response design strategy. Lactic acid and urea formed hydrogen bonds with FF, enriched analyte presence at the heterogeneous interface. Succinic and lactic acids corroded the nZVI passivation layer and promoted electron transfer through surface conjugation. This unique strategy highlights biomolecular coronas as a complex resource to augment nano-enabled technologies and will provide shortcuts for rational manipulation of nanomaterial surfaces with coordinated multifunctionalities.

Water extract of earthworms mitigates kidney injury triggered by oxidative stress via activating intrarenal Sirt1/Nrf2 cascade and ameliorating mitochondrial damage.

ETHNOPHARMACOLOGICAL RELEVANCE: Ischemia-reperfusion (IR) injury can result in acute renal failure. Oxidative stress is a major factor in IR-induced cell death in the kidneys. According to traditional Chinese medicine, earthworms (Pheretima aspergillum) can be used to treat various kidney diseases. AIM OF THE STUDY: The present study was designed to understand the protective effects of the water extract of earthworms (WEE) against oxidative stress on the kidneys and the crucial molecular events associated with its nephroprotective activity. MATERIALS AND METHODS: Cytotoxicity caused by H2O2 in HEK293, HK2, and primary mouse renal tubular epithelial cells (TECs) was used to investigate the effect of WEE on oxidative stress-induced renal injury in vitro. IR-induced kidney injury was established using rats as an in vivo model. The WEE-mediated protection of the kidneys against oxidative stress was compared with that of glutathione, a common antioxidant used as a positive control. RESULTS: In HEK293 cells, HK2 cells, and primary mouse TECs, WEE relieved H2O2-induced mitochondrial damage, apoptosis, and ferroptosis. In kidney cells, WEE increased the expression of Sirt1, boosted LKB1 and AMPK phosphorylation, and upregulated nuclear Nrf2. Suppression of Sirt1 and LKB1 knock down abrogated WEE-induced protection against H2O2. WEE ameliorated IR-induced kidney injury and intrarenal inflammation in rats. In rat kidneys, WEE mitigated mitochondrial damage and suppressed IR-induced apoptosis and ferroptosis. Mechanistically, WEE increased Sirt1 expression, enhanced the phosphorylation of LKB1 and AMPK, and increased intranuclear Nrf2 levels in IR kidneys. IR treatment resulted in considerable increase in renal MDA levels and a prominent decrease in antioxidative enzyme activity. These lesions were significantly alleviated by WEE. CONCLUSIONS: WEE mitigated H2O2-induced cytotoxicity in kidney cells in vitro and improved IR-induced kidney damage in rats. Mechanistically, WEE potentiated the Sirt1/Nrf2 axis and relieved mitochondrial damage in the kidney cells. These events inhibited the apoptosis and ferroptosis induced by oxidative stress. Our findings support the potential application of WEE for the clinical treatment of kidney diseases caused by intrarenal oxidative stress.

Effects of sulfamethazine and tetracycline at molecular, cellular and tissue levels in Eisenia fetida earthworms.

Soil contamination by antibiotics is a global issue of great concern that contributes to the rise of bacterial antibiotic resistance and can have toxic effects on non-target organisms. This study evaluated the variations of molecular, cellular, and histological parameters in Eisenia fetida earthworms exposed to sulfamethazine (SMZ) and tetracycline (TC), two antibiotics commonly found in agricultural soils. The earthworms were exposed for 14 days to a series of concentrations (0, 10, 100, and 1000 mg/kg) of both antibiotics. SMZ and TC did not affect the survival of E. fetida, however, other effects at different levels of biological complexity were detected. The two highest concentrations of SMZ reduced the viability of coelomocytes. At the highest TC concentration, there was a noticeable decline in cell viability, acetylcholinesterase activity (neurotoxicity), and the relative presence of mucopolysaccharides in the epidermis (mucous production). Glutathione S-transferase activity decreased in all TC treatments and at the highest SMZ concentration. However, levels of malondialdehyde and protein carbonyls did not change, suggesting an absence of oxidative stress. Tetracycline was neurotoxic to E. fetida and changed the integrity of the epidermis. Both antibiotics altered the intestinal microbiota of E. fetida, leading to a reduction in the relative abundance of bacteria from the phyla Proteobacteria and Bacteroidetes, while causing an increase in the phylum Actinobacteroidota. All observed changes indicate that both SMZ and TC can disrupt the earthworms' immune system and gut microbiome, while fostering the growth of bacteria that harbour antibiotic resistance genes. Finally, both antibiotics exerted additional metabolic and physiological effects that increased the vulnerability of E. fetida to pathogens.

Grasslands and flood mitigation - Contrasting forages improve surface water infiltration rates.

Grasslands globally deliver many ecosystem services, including water management to alleviate flood risk reduction. Two replicated field experiments were conducted to study how agricultural forage species with diverse rooting systems, sown as single species, affected rooting, soil structure and earthworm populations, and consequently water infiltration to understand how they each might influence flood risk from grasslands. Experiment One showed soils under red clover (Trifolium pratense), white clover (Trifolium repens) and chicory (Cichorium intybus) had higher infiltration rates three years after establishment, compared to perennial ryegrass (Lolium perenne). Higher red clover and chicory root biomass or increased earthworm abundance under white clover may have caused these effects. Experiment Two monitored infiltration at intervals over several years post establishment to understand the timeframe for changes in rates; plantain (Plantago lanceolata) was sown as an additional forage. Infiltration declined post establishment, the timing and extent of decline varying with forages; forage effects were significant after 27 months (P < 0.05). Infiltration rates were higher under red and white clover compared to ryegrass, with chicory and plantain intermediate (P < 0.05). Forages again differed in likely mechanisms delivering higher water infiltration, notably between the two clover species. White clover had higher earthworm biomass (P < 0.05), whereas red clover had a higher average root diameter compared to the other forages (P < 0.05). Drivers of intermediate benefits of chicory and plantain also differed: chicory had higher earthworm abundance (month 38) compared to plantain, which had higher average root diameter compared to ryegrass (month 41); 30 months post-establishment soil bulk density was lower under both forages compared to ryegrass and red clover, with white clover intermediate (P < 0.05); bulk density and penetration resistance did not relate to infiltration. Findings demonstrate that a shift from perennial ryegrass-dominated pastures to swards with more contrasting forages provides an ecohydrological approach to mitigating flood risk and climate adaptation.