Effect of BF839 + earthworm protein supplement on motor and some non-motor symptoms of Parkinson's disease: a randomized clinical trial.

Introduction: Some studies have found that probiotics have the potential to treat PD, and earthworm protein is a traditional Chinese medicine used for the treatment of PD. The purpose of this study was to evaluate the safety and efficacy of Bacteroides fragilis 839 (BF839) + earthworm protein supplement as an adjunctive therapy for PD and to observe changes in the gut microbiota. Methods: Forty-six patients with PD were recruited for a 12-week 1:1 randomized, double-blind, placebo-controlled clinical trial to evaluate changes in motor and some non-motor symptom scores and detect metagenomic changes in the gut microbiota. Results: From baseline to 12 weeks, compared with placebo, the trial group showed significant reductions in the United Parkinson's Disease Rate Scale (UPDRS) total score (-7.74 ± 5.92 vs. -1.83 ± 4.14, p < 0.001), UPDRS part I (-0.72 ± 0.81 vs. -0.20 ± 0.72, p = 0.026), UPDRS part II (-2.50 ± 2.24 vs. -0.22 ± 1.98, p = 0.001), UPDRS part III (-3.43 ± 3.42 vs. -1.33 ± 2.65, p = 0.024), and UPDRS part IV (-1.13 ± 1.19 vs. -0.15 ± 0.57, p = 0.001). Significant reductions in the Hamilton Depression Scale-24 score (-3.91 ± 3.99 vs. +1.15 ± 3.42, p < 0.001), Self-Rating Anxiety Scale scores (-7.04 ± 5.71 vs. -1.23 ± 2.34, p < 0.001), and Constipation scoring system scores (-8.59 ± 4.75 vs. 0.27 ± 1.24, p < 0.001), were also noted. In the trial group, one patient experienced mild eczema and one suffered low blood pressure, which could not be conclusively attributed to supplementation. Compared to the placebo group, the trial group showed a marked increase in Enterococcus faecium and a decrease in Klebsiella. Conclusion: This study is the first to report that probiotics plus earthworm protein can remarkably improve the motor and some non-motor symptoms of PD without serious adverse effects. However, further clinical trials and exploration of the underlying mechanisms are required. Clinical trial registration: Clinical trial registry http://www.chictr.org.cn/, Identification No: ChiCTR2000035122.

Morphophysiological and Histopathological Effects of Ammonium Sulfate Fertilizer on Aporrectodea trapezoides (Dugès, 1828) Earthworm.

The present study used the adult earthworm Aporrectodea trapezoides as a bioindicator species to look into the possible dangers of ammonium sulfate (AS) fertilizer. Two complementary toxicity tests were conducted to determine the LC50values, growth rate inhibition, morphological alterations, and histopathological texture of worms. The lethality test included four increasing concentrations of AS fertilizer (ranging from 2500 to 7500 mg/kg of dry soil weight (d.w.)), while sub-lethal concentrations were based on 10%, 30%, 40%, and 50% of the 14-day median lethal concentration (LC50), with a control group included for both tests. The LC(50) values for AS fertilizer were significantly higher at 7 days (4831.13 mg/kg d.w.) than at 14 days (2698.67 mg/kg d.w.) of exposure. Notably, earthworms exhibited significant growth rate inhibition under exposure to various concentrations and time durations (14/28 exposure days). Morphological alterations such as clitellar swelling, bloody lesions, whole body coiling and constriction, body strangulation, and fragmentation were accentuated steadily, with higher concentrations. Histopathological manifestations included severe injuries to the circular and longitudinal muscular layers, vacuolation, muscle layer atrophy, degradation of the chloragogenous tissue in the intestine, collapsed digestive epithelium of the pharynx with weak reserve inclusion, and fibrosis of blood vessels. These effects were primarily influenced by increasing concentrations of fertilizer and time exposure. The study highlights the strong relationship between concentration and exposure time responses and underscores the potential of A. trapezoides earthworms as valuable biological control agents against acidic ammonium sulfate fertilizer. Importantly, this research contributes to the use of such biomarkers in evaluating soil toxicity and the biological control of environmental risk assessment associated with chemical fertilizers.

Oriented regulation of earthworm production and vermicompost quality by carbon bioavailability management.

Vermicomposting is an efficient bioconversion technology for recycling nutrients from organic waste materials. The biodegradability of raw materials has a significant impact on the earthworm transformation product. However, the management of carbon bioavailability is often overlooked during the vermicomposting process due to the varying degradability of C-rich source in different organic waste. This research aims to investigate the impact of different bioavailable carbon compositions on vermicomposting and to develop a strategy for efficient carbon management. The study involved systematic vermicomposting using four different biodegradable carbon sources (pineapple peels, rice straw, tomato straw, and sawdust) with varying carbon­nitrogen ratios (ranging from 24 to 42). The earthworm production and vermicompost quality were comprehensively evaluated, along with the influence of carbon components on microbial community structure. The results indicated that the optimal vermicomposting treatments were achieved at PCM24, RCM30, TCM30, and MCM30 treatments. Maintaining an approximate ratio of 1:(0.5-1.3) between available and recalcitrant carbon components based on the optimal carbon­nitrogen ratio was found to be optimal for regulating vermicomposting products. Increasing the proportion of available carbon enhanced the quality of vermicompost fertilizer, while a higher proportion of recalcitrant carbon could improve earthworm biomass production efficiency. Labile carbon proportion I (LCP1) and available carbon component (ACC) were identified as key indicators in influencing the formation of microbial community structure. Different carbon compositions led to the specific development and formation of microbial communities, further resulting in significant variations in vermicompost quality under the mediation of microbes. This study, for the first time, clarifies the impact of vermicomposting performance and microbial community from the perspective of carbon bioavailability, which is of great significance for the oriented regulation the vermicomposting efficiency and product in practice.

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.

Earthworms-enhanced bacterial degradation of the chiral fungicide penflufen R-enantiomer.

The widespread application of chiral fungicides as seed-coating agents in agriculture has led to serious residue accumulation in soil, increasingly drawing attention to soil pollution remediation strategies for chiral pesticides. This study explored the role of earthworms and soil microorganisms in selectively accelerating the degradation of penflufen in soil. The results showed that soil microorganisms significantly accelerated penflufen enantiomer degradation, particularly the R-enantiomer. Nocardioides, Variovorax, Arthrobacter, and Pseudomonas were identified as key degrading microorganisms associated with the preferential degradation of the R-enantiomer. The addition of earthworms further significantly enhanced the preferential degradation of the R-enantiomer. Importantly, earthworms markedly promoted the growth and reproduction of the four aforementioned degrading microorganisms in soil treated with enantiomers. Notably, the relative abundance of these degrading microorganisms was significantly higher in R-enantiomer-treated soil with earthworms than in soil treated with the S-enantiomer. Additionally, earthworms significantly increased the relative abundance of degradation genes p450, bphA1, and benA in the soil, especially in the R-enantiomer treated soil. Nocardioides, Variovorax, Arthrobacter, and Pseudomonas were identified as potential hosts for the degradation gene benA. More importantly, twelve strains of penflufen-degrading bacteria were isolated from the treated soil, of which eight belonged to the aforementioned four microorganisms and exhibited a remarkable ability to preferentially degrade the R-enantiomer. This finding highlights the potential of adding earthworms to soil, in conjunction with key degrading microorganisms, which preferentially accelerates penflufen R-enantiomer degradation.

Different mulch films, consistent results: soil fauna responses to microplastic.

Agricultural activities contribute to plastic pollution, with unintentional introduction and intentional use of plastic mulch films leading to the accumulation of microplastic particles in soils. The lack of removal techniques and scarce information on the effects on soil organisms, especially for biodegradable mulch films, necessitate an assessment of potential effects. This study aimed to elucidate the effects of mulch film microplastic on soil fauna by investigating reproduction output and subcellular responses before and after recovery from exposure. Two common soil organisms, Folsomia candida and Eisenia fetida, were exposed to petroleum-based polyethylene (PE) and biodegradable polylactic acid/polybutylene adipate terephthalate (PLA/PBAT) microplastic for 28 days, according to OECD guidelines 232 and 222, respectively. Juvenile numbers revealed no polymer- or concentration-dependent effects on E. fetida and F. candida reproduction after exposure to up to 5 and 10 g/kgdw soil, respectively. To provide a more sensitive and early indication of sublethal effects, subcellular responses in E. fetida were analyzed. Glutathione S-transferase (GST) activity increased with rising microplastic concentration; however, catalase (CAT), acetylcholine esterase (AChE) activity, and reactive oxygen species (ROS) did not differ from control levels. Further, the more environmentally relevant PE polymer was chosen for in-depth assessment of subcellular response after 28-day microplastic exposure and subsequent 28 days in uncontaminated soil with E. fetida. No significant differences in biomarker activity and stress levels were observed. We conclude that mulch film-derived microplastic did not adversely affect earthworm and collembolan species in this scenario, except for a slight induction in the detoxification enzyme glutathione S-transferase.

Conservation agriculture practices impact on biological and microbial diversity in earthworm cast under maize-wheat system.

Soil degradation is a major global concern due to its negative impact on soil quality and the sustainability of agricultural resources. The conservation agriculture (CA) approach, which includes three key principles such as zero tillage, retention of crop residue and crop rotation has gained widespread adoption to help mitigate the climate change effects on agricultural soils and meet the growing demand for increased production. Earthworm communities, along with microbial activity and diversity, are highly sensitive to tillage practices. Additionally, microbial activity and diversity quickly respond to different cropping systems, making them effective indicators for detecting short-term changes in soil functioning. We therefore, assess the effects of CA innovative approached after 6-years on biological and microbial diversity within earthworm cast in maize-wheat system (MWS). The treatments consist of PBM-RN0/ZTW-RN0 (permanent beds No-N control-both residues removed and wheat with zero tillage); PBM+RN0/ZTW+RN0 (permanent beds No-N control-both residues retained)-50% of maize stover and 25% of wheat residue retained; PBM-RN120/ZTW- RN120 (permanent beds with 120 kg N ha-1 both residues removed wheat with zero tillage); PBM+RN120/ZTW+RN120 (permanent beds with 120 kg N ha-1 both residues retained and wheat with zero tillage) and FBM-RN120/CTW-RN120 (fresh beds in maize/CT in wheat with 120 kg N ha-1 both residues removed). The result of present study showed that activities of carbon (C) cycle-related enzymes in the cast soils viz., dehydrogenase (DHA), ß-glucosidase (ß-glu), cellulase, and xylanase were significantly higher under PBM+RN120/ ZTW+RN120 than under PBM-RN0/ZTW-RN0. Specifically, the activities of these enzymes were 21.5, 26.8, and 76.5% higher under the PBM+RN120/ZTW+RN120 treatment, respectively. Moreover, the Alk-P activity was found to be 1.3 times higher in the PBM+RN120/ZTW+RN120 treatment than in the PBM-RN0/ZTW-RN0 treatment. The bacterial, fungal, and actinomycete counts in the cast soil ranged from 6.87 to 7.47 CFU (colony forming units) x 106 g-1 soil, 3.87-3.30 CFU x 104 g-1 soil, and 5.09-5.67 CFU x 104 g-1 soil, respectively. Total organic carbon (TOC) showed significant increases of 34.6% under PBM+RN120/ZTW+RN120 as compared to PBM-RN0/ZTW-RN0. The less labile C (Frac. 3), total carbohydrate carbon (TCHO), phenol oxidase (PHE) and peroxidase (PER) were observed as the sensitive indicators under different tillage, rate of nitrogen and residue management practices. This study suggests that permanent beds with crop residue retention with balance fertilization practices can be recommended and popularized to the overall improvement of soil biological pools within earthworm casts in MWS.

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.

The effect of soil parameters and earthworm abundance on the fine-scale nocturnal habitat use of the Eurasian woodcock (Scolopax rusticola).

The Eurasian woodcock prefers habitats where its main prey, earthworms, can be found in higher densities. Although they are forest-dwelling birds, they regularly visit pastures and natural grasslands at night, where earthworm abundance is generally higher. However, there is little information on fine-scale habitat use in relation to variation in habitat characteristics and prey availability, particularly beyond the breeding season. In our study, we investigated if the nocturnal occurrence of woodcocks during migratory stopover periods differed between two neighbouring fields, or management units, with similar vegetation structure, and if within-field variation in the spatial patterns of woodcock sightings were associated with fine-scale earthworm densities and soil parameters. Specifically, we used GPS tracking data of two tagged woodcocks and direct observation data to study patterns of occurrence of birds in a mixed forest-pasture landscape in Hungary during pre- and post-breeding periods. We compared these patterns with fine-scale soil characteristics and earthworm abundance, acquired by field sampling. We found that the field with higher earthworm abundance was visited by woodcocks more frequently, and this correlation was similarly observed at the intra-field level. Our results demonstrate that woodcocks select foraging sites with higher earthworm densities at multiple spatial scales, both between fields (coarse scale), and within fields (fine-scale). Considering that woodcocks tended to return to the same field to forage at night, the strong associations between occupancy and resources provide a basis for developing habitat management strategies at the field level for conservation. As earthworm densities and soil parameters are good indicators of woodcock foraging habitat, measuring those variables, at least at a coarse scale, could aid in predicting important habitats for the species across the landscape.

Advancing Soil Risk Assessment: A Novel Earthworm Cocoon Test with a Complementary Toxicokinetic-Toxicodynamic Modeling Approach.

In the current European Union pesticide risk assessment for soil organisms, effect endpoints from laboratory studies (Tier 1) and field studies (higher-tier risk assessment) are compared with predicted environmental concentrations in soil, derived from the proposed use pattern. The simple but conservative initial Tier 1 risk assessment considers a range of worst-case assumptions. In contrast, the higher-tier assessment focuses on specific conditions tested in the corresponding field study. Effect modeling, such as toxicokinetic-toxicodynamic (TKTD) modeling, is considered a promising future tool to address uncertainties in soil risk assessment, such as extrapolation to different ecological, pedo-climatical, or agronomical situations, or to serve as an intermediate tier for potential refinement of the risk assessment. For the implementation of TKTD modeling in soil organism risk assessment, data on earthworm growth and reproduction over time are required, which are not provided by the standard Organisation for Economic Co-operation and Development (OECD) 222 laboratory test. The underlying study with carbendazim presents a new earthworm cocoon test design, based on the OECD 222 test, to provide the necessary data as input for TKTD modeling. This proposed test design involves destructive samplings at days 7, 14, 21, and 28, enabling the determination of growth, cocoon number, and the number of juveniles hatched per cocoon in 7-day intervals. The new cocoon test allowed the disentanglement of the toxic effect of carbendazim in earthworms: At the highest concentration prominent effects on growth and reproductive output were observed, and the number of cocoons was significantly reduced compared to control. The results highlighted different physiological modes of action: effect on growth via higher maintenance costs as a primary mode of action as well as a reduced number of cocoons (effect on reproduction) and a lower number of juveniles hatching from each cocoon (hazard during oogenesis) as a secondary mode of action. We provide an example of how this new test's data can be used to feed a dynamic energy budget theory-TKTD model of Eisenia fetida. We also validate it against the original OECD 222 test design, outlining its potential future use in soil risk assessment. Environ Toxicol Chem 2024;00:1-10. © 2024 The Author(s). Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

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.

Comparative Analysis of Bacterial Community Structures in Earthworm Skin, Gut, and Habitat Soil across Typical Temperate Forests.

Earthworms are essential components in temperate forest ecosystems, yet the patterns of change in earthworm-associated microbial communities across different temperate forests remain unclear. This study employed high-throughput sequencing technology to compare bacterial community composition and structure in three earthworm-associated microhabitats (skin, gut, and habitat soil) across three typical temperate forests in China, and investigated the influence of environmental factors on these differential patterns. The results indicate that: (1) From warm temperate forests to cold temperate forests, the soil pH of the habitat decreased significantly. In contrast, the physicochemical properties of earthworm skin mucus exhibited different trends compared to those of the habitat soil. (2) Alpha diversity analysis revealed a declining trend in Shannon indices across all three microhabitats. (3) Beta diversity analysis revealed that the transition from warm temperate deciduous broad-leaved forest to cold temperate coniferous forest exerted the most significant impact on the gut bacterial communities of earthworms, while its influence on the skin bacterial communities was comparatively less pronounced. (4) Actinobacteria and Proteobacteria were the predominant phyla in earthworm skin, gut, and habitat soil, but the trends in bacterial community composition differed among the three microhabitats. (5) Mantel tests revealed significant correlations between bacterial community structures and climatic factors, physicochemical properties of earthworm habitat soil, and physicochemical properties of earthworm skin mucus. The findings of this study offer novel perspectives on the interplay between earthworms, microorganisms, and the environment within forest ecosystems.

Metabolomics and microbiomics revealed the combined effects of different-sized polystyrene microplastics and imidacloprid on earthworm intestinal health and function.

The coexistence of pesticides and plastic film residues in agricultural soils poses a significant threat to soil organisms due to their potential long-term contamination and combined toxic effects. Specifically, earthworms are at risk of simultaneously ingesting residual pesticides and microplastics, yet the impact of this combined exposure on their intestinal health and function remains poorly understood. In this study, earthworm (Eisenia fetida) were single and combined exposed to three particle sizes (10 µm, 500 µm, and 2 mm) of polyethylene microplastics (PE MPs) and imidacloprid (IMI) for 28 days, respectively. Our findings underscore that compared to single exposures, the combined exposure inflicted more profound injuries on intestinal tissues and elicited a heightened activation of intestinal digestive enzymes. Furthermore, the combined exposure significantly perturbed the relative abundance of several pivotal metabolic-associated gut microbiota, fostering an enrichment of pathogenic species. Metabolomics analysis showed combined exposure increased differential metabolites, disrupting amino acid, fatty acid, and carbohydrate metabolism in earthworm intestines, potentially hindering nutrient absorption and causing toxic metabolite accumulation. An integrated omics analysis implies that combined exposures have the potential to disrupt the relative abundance of crucial gut microbiota in earthworms, thereby altering their intestinal metabolism and subsequently impacting intestinal health and functionality. Overall, the results reveal that combined exposure of IMI and PE MPs exacerbate the negative effects on earthworm gut health, and this study holds significant implications for the holistic understanding of the combined toxic effects of microplastics and pesticide on soil ecosystems.

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.

Effect of Insecticide Exposure Across Multiple Generations of the Earthworm Eisenia andrei.

The toxicity of neonicotinoids and many of their replacement insecticides to nontarget soil invertebrates such as earthworms has previously been established. However, the long-term effects of these substances on these organisms are largely unknown. In the field of soil ecotoxicology, lumbricid earthworms such as Eisenia andrei are used extensively due to the availability of standardized test methods and their adaptability to laboratory culture and testing. Multigenerational studies have gained popularity and attention in recent years, with a shift toward the use of long-term assays and lower concentrations of test chemicals. The use of exposure concentrations that include those measured in a monitoring program carried out by the Government of Ontario presents a realistic exposure scenario that may not show significant effects in contemporary, shorter term studies. We used current standardized test methods as a basis for the development of multigenerational studies on E. andrei. The effects of exposure to a single application of the insecticides thiamethoxam and cyantraniliprole on the survival and reproduction of E. andrei were observed over three (thiamethoxam) or two (cyantraniliprole) generations using consecutive reproduction tests. No significant impacts on adult survival were reported in any generation for either insecticide, whereas reproduction decreased between the first and second generations in the thiamethoxam test, with median effective concentration (EC50) values of 0.022 mg/kg dry weight reported for the first generation compared with 0.002 mg/kg dry weight in the second generation. For cyantraniliprole, an EC50 of 0.064 was determined for the first generation compared with 0.016 mg/kg dry weight in the second generation. A third generation was completed for the thiamethoxam test, and a significant decrease in reproduction was observed in all treatments and controls compared with previous generations. No significant difference between thiamethoxam treatments and the control treatment was reported for the third generation. Collectively, these data indicate that exposure of oligochaetes to these two insecticides at concentrations representative of field conditions may result in long-term stresses. Environ Toxicol Chem 2024;43:2058-2070. © 2024 The Author(s). Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Accumulation of heavy metals from single and combined olive mill wastewater and pomace in soil and bioaccumulation in tissues of two earthworm species: Endogeic (Aporrectodea trapezoides) and Epigeic (Eisenia fetida).

Soil and earthworms are threatened by anthropogenic contamination resulting from olive mill waste dumping on the soil due to their pollutant properties. While several studies have explored the effects of olive mill waste on soil properties and the accumulation of heavy metals in soil, there is currently a gap in the literature regarding the potential bioaccumulation of heavy metals from olive mill waste in earthworms. In this study, soil with earthworms from two ecological categories (endogeic: Aporrectodea trapezoides and epigeic: Eisenia fetida) was treated with increasing doses of olive mill wastewater (OMWW) and olive mill pomace (OMP), applied individually or combined, in an indoor experiment in plastic containers, under laboratory conditions. The results revealed the presence of significant concentrations of heavy metals in the two types of wastes ranging as follows: Fe˃ Zn˃ Cu˃ Cd˃ Cr for OMWW, and Fe˃ Zn˃ Cu˃ Cr for OMP (with Cd below the detection limit). The study demonstrated distinct effects of OMWW and OMP, both individually and in combination, on soil heavy metal content, ranging as follows: soil OMWW > soil Combination > soil OMP for Cd; soil Combination > soil OMWW > soil OMP for Cr and Fe; and soil Combination > soil OMP > soil OMWW for Cu and Zn. Additionally, our investigation showed that both earthworm species exhibited significant uptake of these metals into their tissues, particularly the endogeic species. Interestingly, the most significant difference between species was in the accumulation of Cu, with the epigeic species accumulating significantly lower amounts.

Cultivation of earthworms and analysis of associated bacterial communities during earthworms' growth using two types of agricultural wastes.

Earthworm cultivation can effectively promote the resource utilization of agricultural waste. The efficient utilization of agricultural waste by earthworms mainly depends on the microbial communities in the guts. This study used silkworm excrement and cow manure as substrates for earthworm cultivation and investigated the associated bacterial communities during earthworms' growth. The survival rate of earthworms remained above 89% after 21 days of feeding with the two substrates. Proteobacteria, Actinobacteria, and Firmicutes constituted the predominant bacterial communities in earthworm growth, accounting for over 81% of the relative abundance in both guts and vermicompost. The bacteria richness and diversity in the foregut and midgut of earthworm were lower than those in the hindgut. The prediction function of intestinal bacterial communities of earthworms cultured with two substrates mainly involved biosynthesis, decomposition and energy production.

Pharmacological effects of bioactive agents in earthworm extract: A comprehensive review.

This review compiles information from the literature on the chemical composition, pharmacological effects, and molecular mechanisms of earthworm extract (EE) and suggests possibilities for clinical translation of EE. We also consider future trends and concerns in this domain. We summarize the bioactive components of EE, including G-90, lysenin, lumbrokinase, antimicrobial peptides, earthworm serine protease (ESP), and polyphenols, and detail the antitumor, antithrombotic, antiviral, antibacterial, anti-inflammatory, analgesic, antioxidant, wound-healing, antifibrotic, and hypoglycemic activities and mechanisms of action of EE based on existing in vitro and in vivo studies. We further propose the potential of EE for clinical translation in anticancer and lipid-modifying therapies, and its promise as source of a novel agent for wound healing and resistance to antibiotic tolerance. The earthworm enzyme lumbrokinase embodies highly effective anticoagulant and thrombolytic properties and has the advantage of not causing bleeding phenomena due to hyperfibrinolysis. Its antifibrotic properties can reduce the excessive accumulation of extracellular matrix. The glycolipoprotein extract G-90 can effectively scavenge reactive oxygen groups and protect cellular tissues from oxidative damage. Earthworms have evolved a well-developed defense mechanism to fight against microbial infections, and the bioactive agents in EE have shown good antibacterial, fungal, and viral properties in in vitro and in vivo experiments and can alleviate inflammatory responses caused by infections, effectively reducing pain. Recent studies have also highlighted the role of EE in lowering blood glucose. EE shows high medicinal value and is expected to be a source of many bioactive compounds.

Feasibility of composted green waste amended by vermiculite and earthworm casts as the growth media for three common ornamental plants.

This study demonstrated the effects of adding specific proportions of vermiculite (VMT: 0%, 10%, and 20%) and earthworm casts (EWCs: 0%, 10%, and 20%) on the physico-chemical properties of composted green waste (CGW), and the impacts of amended CGW as growth media on the growth of three common ornamental plants (Dahlia pinnata Cav. [dahlia], Centaurea cyanus L. [cornflower], and Consolida ajacis [L.] Schur [delphinium]). Compared with Treatment T1 (CK), the addition of 10% VMT and 20% EWCs greatly (p < 0.05) increased the total porosity, aeration porosity, water-holding porosity, total nitrogen, available phosphorus, available potassium, and organic matter of CGW by 9%, 35%, 4%, 18%, 27%, 13%, and 33%, respectively. In addition, this pattern increased (p < 0.05) the total fresh biomass, total chlorophyll content, and root length of dahlias by 9%, 19%, and 27%, respectively; those of cornflowers by 17%, 30%, and 29%, respectively (p < 0.05); and those of delphiniums by 23%, 14%, and 63%, respectively. Therefore, the amended CGW supplemented with 10% VMT and 20% EWCs was an ideal growth medium for the three plants.

Evaluation of immunotoxicity of iron oxide nanoparticles on coelomocytes of Eisenia fetida.

Iron oxide nanoparticles (Fe3O4 NPs) have gained considerable attention due to their diverse applications in various fields. However, concerns about their potential toxic effects on the environment and living organisms have also emerged. In this study, we synthesized and characterized Fe3O4 NPs and assessed their immunotoxicity on the coelomocytes of Eisenia fetida. The Fe3O4 NPs were synthesized using a co-precipitation method, and their physicochemical properties were determined using techniques such as X-ray diffraction (XRD), scanning electron microscopy-energy dispersive X-ray (SEM-EDX), transmission electron microscopy (TEM) and Fourier-transform infrared spectroscopy (FTIR). The synthesized Fe3O4 NPs exhibited a uniform size distribution with spherical morphology and the phase purity was confirmed from XRD analysis. To evaluate the immunotoxicity of Fe3O4 NPs, Eisenia fetida coelomocytes were exposed to various concentrations of Fe3O4 NPs for 14 days. Furthermore, we analyzed the impact of Fe3O4 NPs on the biochemical parameters, including superoxide dismutase (SOD), catalase (CAT), acid phosphatase (APs), alkaline phosphatase (ALP), and total protein content (TPC), as well as conducted a histological examination. Biochemical analysis revealed significant alterations in the activity levels of SOD, CAT, APs, ALP, and TPC in the coelomocytes, indicating immune system dysregulation upon exposure to Fe3O4 NPs. Moreover, histological examination demonstrated structural changes, suggesting cellular damage caused by Fe3O4 NPs. These findings provide valuable insights into the immunotoxic effects of Fe3O4 NPs on Eisenia fetida and underscore the need for further investigation into the potential environmental impact of nanoparticles.