Application of earthworm can enhance biological power generation and accelerate sulfamethoxazole removal in agricultural soils.

Microbial degradation plays a crucial role in removing sulfonamides from soil, enhancing sulfamethoxazole (SMX) remediation. To further augment SMX removal efficiency and mitigate the transmission risk associated with antibiotic resistance genes (ARGs), this study proposes a novel approach that integrates micro-animals, microorganisms, and microbial fuel cell (MFC) technology. The results showed that earthworm-MFC synergy substantially reduces SMX content and ARGs abundance in soil. The introduction of earthworms enhances humus content, facilitating electron transfer within MFC and consequently improving current generation. Furthermore, electrical stimulation applied to earthworms led to increased protein secretion and enhanced antioxidant system activity, thereby accelerating SMX degradation. Earthworms also foster MFC-associated bacterial growth and SMX-degrading bacteria proliferation, augmenting MFC treatment efficacy. This synergistic effect significantly augmented the overall efficacy of MFC treatment for antibiotics. Overall, integrating earthworm activity with MFC technology effectively optimizes electricity generation and enhances pollutant removal.

Oxidative stress and DNA alteration on the earthworm Eisenia fetida exposed to four commercial pesticides.

Modern agriculture is mainly based on the use of pesticides to protect crops but their efficiency is very low, in fact, most of them reach water or soil ecosystems causing pollution and health hazards to non-target organisms. Fungicide triazoles and strobilurins based are the most widely used and require a specific effort to investigate toxicological effects on non-target species. This study evaluates the toxic effects of four commercial fungicides Prosaro® (tebuconazole and prothioconazole), Amistar®Xtra (azoxystrobin and cyproconazole), Mirador® (azoxystrobin) and Icarus® (Tebuconazole) on Eisenia fetida using several biomarkers: lipid peroxidation (LPO), catalase activity (CAT), glutathione S-transferase (GST), total glutathione (GSHt), DNA fragmentation (comet assay) and lysozyme activity tested for the first time in E. fetida. The exposure to Mirador® and AmistarXtra® caused an imbalance of ROS species, leading to the inhibition of the immune system. AmistarXtra® and Prosaro®, composed of two active ingredients, induced significant DNA alteration, indicating genotoxic effects. This study broadened our knowledge of the effects of pesticide product formulations on earthworms and showed the need for improvement in the evaluation of toxicological risk deriving from the changing of physicochemical and toxicological properties that occur when a commercial formulation contains more than one active ingredient and several unknown co-formulants.

Phylogenetic Illustration of Eisenia fetida Associated Vermi-bacteria Involved in Heavy Metals Remediation and Retaining Plant Growth Promoting Traits.

Heavy metals contamination in the soil is a major threat to wildlife, the environment, and human health. Microbial remediation is an emerging and promising technology to reduce heavy metals toxicity. Therefore, the present research aimed to isolate and to identify the heavy metals tolerated bacteria from the Eisenia fetida for the first time, and to screen the bacto-remediation capabilities and plant growth promoting traits of vermi-bacterial isolates. Vermi-bacteria was isolated from the gut of E. fetida, identified through staining, culturing, biochemical tests, and ribotyping. Plant growth-promoting traits were also evaluated. Phylogenetic results revealed that isolated Vermi-bacterial strains showed resemblance with Bacillus thuringiensis, Bacillus aryabhattai, Staphylococcus hominis, Bacillus toyonensis, Bacillus cabrialesii, Bacillus tequilensis, Bacillus mojavensis, Bacillus amyloliquefaciens, Bacillus toyonensis, Bacillus anthracis, and Bacillus paranthracis. All identified Vermi-bacterial species are Gram-positive (rod and cocci) in nature, not only indicated the efficient biosorption of lead, cadmium, and chromium but also produce all plant growth stimulating traits such as indole acetic acid (IAA), amylase, protease, lipase, hydrogen cyanide, ammonia, and siderophore production, and also act as a phosphate solubilizers. Bacillus anthracis showed significant production of siderophore (33.0±0.0 mm), phosphate solubilizing (33.0±0.0 mm), proteolytic (15.0±0.0 mm), and lipolytic activities (20.0±0.0 mm) compared to other vermi-bacterial isolates. Bioaccumulation factor results revealed that Bacillus anthracis showed more accumulation of Cd (12.00±0.01 ppm), Cr (5.38±0.01 ppm), and Pb (4.38±0.01 ppm). Therefore, the current findings showed that all identified vermi-bacteria could be used as potential bactoremediation agents in heavy metals polluted environments and could be used as microbial biofertilizers to enhance crop production in a polluted area.

Vermicomposting of invasive weed Ageratum conyzoids: Assessment of nutrient mineralization, enzymatic activities, and microbial properties.

Ageratum conyzoids biomass was vermicomposted with cow dung in 25 (T25), 50 (T50), and 75 (T75) % (v/v) ratios and changes in physicochemical characteristics, enzymatic activities (proteases, dehydrogenases, ß-galactosidase and phosphatases), and microbial population (bacterial, fungal, and actinomycetes) was recorded. Vermicomposting caused a decrease in pH, OCtotal (27.3-35.3%), but an increase in Ntotal (59.6-69.9%), Paval (53.8-148.7%), Ktotal (32.2-92.43%), and Catotal (25.5-55.3%). The peaked enzymatic activities were recorded between 5 and 15 d. T50 and T75 showed the highest fold increase in bacteria (2.09-to-2.51), fungi (1.48-to-2.41), actinomycetes (1.52-to-1.79) population. The maximum biomass (883.67-1480 mg), cocoon production (85.33-145.33), and population build-up in earthworm were recorded in setups with a high content of Ageratum. Germination index (>80%) and soil respiration rate suggested the non-toxic impact of vermicomposted Ageratum. Results indicate that Ageratum could be biotransformed into toxic-free manure through vermitechnology.

Bioremediation of diesel and gasoline-contaminated soil by co-vermicomposting amended with activated sludge: Diesel and gasoline degradation and kinetics.

Present study aims to examine the efficiency of co-vermicomposting amended with activated sludge and E. fetida earthworm for bioremediation of diesel and gasoline from contaminated soil. The diesel and gasoline removal efficiency and degradation rates coefficients were estimated with gas chromatography (GC) analysis and first-order kinetics. The removal of gasoline and diesel in different co-vermicomposting processes with and without E. fetida ranged between 65-100% and 24.94-63.93%, respectively within 90- day experiment. Removal of gasoline and diesel increased in soil with addition of earthworm (E. fetida); higher degradation rate coefficients (k) were observed for co-vermicomposting with earthworm compared with co-vermicomposting processes. The highest k (0.014) for diesel degradation was estimated for microcosm reactor 4 (R4), where high numbers of E. fetida accelerate the less biodegradable organic contaminant from the soil matrices. The reasonable survival rates of earthworms in exposure to high concentration of petroleum-derivatives contaminated soils indicated increased activity of ligninolytic diesel-degrading earthworms and microorganisms. Therefore, co-vermicomposting amended with activated sludge is suggested as feasible and promising technologies for bioremediation of high content of organic contaminants from the soil matrices.

Histopathological effects and biomarker response of earthworms, Eisenia fetida, after exposure to crude oil contaminated soils.

Earthworms are the most abundant invertebrates in the soils and are permanently in close contact with soil particles. Therefore, they are significantly affected by the pollutants that reach the soil system. The study was aimed at evaluating the effect of exposure to crude oil contaminated soil on Eisenia fetida using cellular antioxidant enzymes and tissue organization as biomarkers. E. fetida were exposed to different concentrations of crude oil contamination of 1 mL, 2 mL. and 3 mL (0.25, 0.50 and 0.75%) for 14 days. The antioxidant/oxidant parameters were analysed in the muscle and liver tissues. The results showed that only the reduced glutathione (GSH) of earthworms exposed to 1 mL were not significant (p>0.05) from the control, while the other concentrations (2 mL - 0.50% and 3 mL - 0.75%) were significantly different (p<0.05) from the control. The activity of catalase (CAT) with respect to the total protein content was highest in the liver of earthworms exposed to 1 mL (0.25% conc.) on day 7 (51.84 µ mol/mg pro) while the least CAT activity with respect to the total protein content was reported in the liver of control earthworms after 14 days (19.51 µ mol/mL/min). A significant increase in the activity of superoxide dismutase at all the concentrations (0.25, 0.50 and 0.75%) after 14 days were also observed. Significant histopathological alterations were observed in E. fetida from the three concentrations. Severe disruptions in the arrangement of their body wall muscle layers, distorted internal viscera, as well as cellular degeneration, pigments, moderate to severe areas of lesion, and distortion of the shape of circular and longitudinal muscles, eroding of internal and external tissues leading to total destruction of body wall were observed. In conclusion, the study revealed that crude oil even at lower concentration induced biomarker responses in E. fetida such as higher levels of Malondialdehyde in E. fetida after exposure crude oil due its toxicity. Histopathological alterations such as cellular degeneration, moderate to severe areas of necrosis, areas of inflammation, inclusion bodies, pigments, and distortion of the shapes of circular and longitudinal muscles also showed the adverse impacts of crude oil pollution in the soils.

Recycling of lignocellulosic waste as vermicompost using earthworm Eisenia fetida.

Present study aimed to evaluate the vermicomposting of lignocellulosic wastes employing Eisenia fetida earthworms. The study examined the effectiveness of vermicomposting for 105 days by mixing lignocellulosic waste (LW) with cattle manure (CM) in five different proportions. Results revealed that TOC and C/N ratio decreased gradually till end and in vermicomposts varied between 268-320 g/kg and 12.26-16.85, respectively. Nutrient content (NPK) in the vermicomposts increased with time in all the mixtures. Heavy metals' content also increased in vermicomposts and benefit ratio for heavy metals ranged between 0.06 and 5.1. Increase in earthworm biomass (22.38-39.64 g) and reproduction (21.27-31.60 hatchlings/worm) was also satisfactory in all the waste mixtures. Based on results, it can be inferred that lignocellulosic waste can successfully be converted into good quality manure employing earthworms.

Effects of soil properties and aging process on the acute toxicity of cadmium to earthworm Eisenia fetida.

This study was undertaken to investigate the effects of soil properties and aging process on the acute toxicity of cadmium (Cd) to Eisenia fetida (E. fetida) in 18 Cd-spiked soils. Results showed that the Cd toxicity to E. fetida differed in the 18 soils with different characteristics, and median lethal concentration (LC50) values varied from 440.7 to 1520.4 mg/kg in freshly spiked soils. Soil pH and organic matter (OM) content were the two major factors associated with Cd toxicity. The increase in LC50 values and decreases in both exchangeable Cd in soils and tissue Cd concentrations in earthworm whole body indicated that aging (180 and 360 days) could reduce the acute toxicity and bioavailability of Cd to E. fetida. Cadmium concentrations in E. fetida were positively correlated with exchangeable Cd content in soils, and soil pH and OM were the key factors controlling the distribution and transformation of the exchangeable Cd. The results will provide useful reference information for the risk assessment of Cd in the terrestrial environment.

De novo transcriptome assembly, functional annotation and differential gene expression analysis of juvenile and adult E. fetida, a model oligochaete used in ecotoxicological studies.

BACKGROUND: Earthworms are sensitive to toxic chemicals present in the soil and so are useful indicator organisms for soil health. Eisenia fetida are commonly used in ecotoxicological studies; therefore the assembly of a baseline transcriptome is important for subsequent analyses exploring the impact of toxin exposure on genome wide gene expression. RESULTS: This paper reports on the de novo transcriptome assembly of E. fetida using Trinity, a freely available software tool. Trinotate was used to carry out functional annotation of the Trinity generated transcriptome file and the transdecoder generated peptide sequence file along with BLASTX, BLASTP and HMMER searches and were loaded into a Sqlite3 database. To identify differentially expressed transcripts; each of the original sequence files were aligned to the de novo assembled transcriptome using Bowtie and then RSEM was used to estimate expression values based on the alignment. EdgeR was used to calculate differential expression between the two conditions, with an FDR corrected P value cut off of 0.001, this returned six significantly differentially expressed genes. Initial BLASTX hits of these putative genes included hits with annelid ferritin and lysozyme proteins, as well as fungal NADH cytochrome b5 reductase and senescence associated proteins. At a cut off of P = 0.01 there were a further 26 differentially expressed genes. CONCLUSION: These data have been made publicly available, and to our knowledge represent the most comprehensive available transcriptome for E. fetida assembled from RNA sequencing data. This provides important groundwork for subsequent ecotoxicogenomic studies exploring the impact of the environment on global gene expression in E. fetida and other earthworm species.

Nutrient changes and biodynamics of Eisenia fetida during vermicomposting of water lettuce (Pistia sp.) biomass: a noxious weed of aquatic system.

This paper reports the results of vermicomposting of water lettuce biomass (WL) spiked with cow dung at ratios of 20, 40, 60, and 80 % employing Eisenia fetida. A total of four treatments were established and changes in chemical properties of mixtures were observed. Vermicomposting caused a decrease in pH, TOC, volatile solids, and C/N ratio by 1.01-1.08-fold, 0.85-0.92-fold, 0.94-0.96-fold, 0.56-0.70-fold, respectively, but increase in EC, totN, totP, totK, totCa, totZn, totFe, and totCu, by 1.19-1.42-fold, 1.33-1.68-fold, 1.38-1.69-fold, 1.13-1.24-fold, 1.04-1.11-fold, 1.16-1.37-fold, 1.05-1.113-fold, 1.10-1.27-fold, respectively. Overall, the treatment with 60-80 % of WL showed the maximum decomposition and mineralization rates. The earthworm showed the growth and reproduction rate in considerable ranges in all treatment setups but setups with 60-80 % WL proportion exhibited the optimum results. Results reveal that biomass of water lettuce can be utilized effectively for production of valuable manure through vermicomposting system.

De novo transcriptome assembly, functional annotation and differential gene expression analysis of juvenile and adult E. fetida, a model oligochaete used in ecotoxicological studies

BACKGROUND: Earthworms are sensitive to toxic chemicals present in the soil and so are useful indicator organisms for soil health. Eisenia fetida are commonly used in ecotoxicological studies; therefore the assembly of a baseline transcriptome is important for subsequent analyses exploring the impact of toxin exposure on genome wide gene expression. RESULTS: This paper reports on the de novo transcriptome assembly of E. fetida using Trinity, a freely available software tool. Trinotate was used to carry out functional annotation of the Trinity generated transcriptome file and the transdecoder generated peptide sequence file along with BLASTX, BLASTP and HMMER searches and were loaded into a Sqlite3 database. To identify differentially expressed transcripts; each of the original sequence files were aligned to the de novo assembled transcriptome using Bowtie and then RSEM was used to estimate expression values based on the alignment. EdgeR was used to calculate differential expression between the two conditions, with an FDR corrected P value cut off of 0.001, this returned six significantly differentially expressed genes. Initial BLASTX hits of these putative genes included hits with annelid ferritin and lysozyme proteins, as well as fungal NADH cytochrome b5 reductase and senescence associated proteins. At a cut off of P = 0.01 there were a further 26 differentially expressed genes. CONCLUSION: These data have been made publicly available, and to our knowledge represent the most comprehensive available transcriptome for E. fetida assembled from RNA sequencing data. This provides important groundwork for subsequent ecotoxicogenomic studies exploring the impact of the environment on global gene expression in E. fetida and other earthworm species.

Uptake route and resulting toxicity of silver nanoparticles in Eisenia fetida earthworm exposed through Standard OECD Tests.

Despite the increasing interest in silver nanoparticles toxicity still few works dealt with the hazards of nanosized Ag in soils (either dissolved in pore water or coupled to colloids) although disposal of biosolids in landfills has been reported as the major source of silver nanoparticles in terrestrial environments. Presently, Eisenia fetida was used to assess the toxicity of 5 nm sized PVP-PEI coated silver nanoparticles in soil through the implementation of different exposure media Standard Toxicity Tests (Paper Contact and Artificial Soil -OECD-207- and Reproduction -OECD-222- Tests) together with cellular biomarkers measured in extruded coelomocytes. In order to decipher the mode of action of silver nanoparticles in soil and the uptake routes in earthworms, special attention was given to the Ag accumulation and distribution in tissues. High Ag accumulation rates, weight loss, and mortality due to the disruption of the tegument could be the result of a dermal absorption of Ag ions released from silver nanoparticles (Paper Contact Test). However, autometallography showed metals mainly localized in the digestive tract after Artificial Soil Test, suggesting that Ag uptake occurred mostly through soil ingestion. That is, silver nanoparticles attached to soil colloids seemed to be internalized in earthworms after ingestion of soil and transferred to the digestive gut epithelium where at high doses they have triggered severe effects at different levels of biological complexity.

Adaptability comparison of E. fetida in vermicomposting against sludge from livestock wastewater treatment plant based on their several growth stages.

Vermicomposting is a low-cost, eco-efficient process to deal with organic wastes. Mixtures of swine manure (SM), cow dung (CD), and animal wastewater treatment plant sludge (S) were applied as feeds, and Eisenia fetida was employed in this study to investigate the vermicomposting efficiency based on their several growth stages. The hatching test resulted in a 100 % hatching rate in S60SM40 (60 % S + 40 % SM) mixture, while 4.40 hatchlings per cocoon were observed. The growth of infancy performed best in 0-20 % CD mixtures (0.05 ± 0.002 g), followed by in SM + CD (0.04 ± 0.003 g). The highest growth rate of young and adult E. fetida was noticed in CD + S mixtures (11.14 ± 0.01 and 6.00 ± 0.02 mg/d/worm, respectively), while the higher cocoon production of adults was noticed in S + SM mixtures especially in S40SM60 (537 ± 5 worms). Moreover, the conversion of solids; the modified pH value; the reduction in total organic carbon (TOC); total Kjeldahl nitrogen (TKN), NH4-N, NO3-N, and C:N ratio; and the rich in total available phosphorus (TAP) and total potassium (TK) content by young and adult E. fetida were related to the growth of worms. Such work would benefit understanding and to increase the efficiency of vermicompost processing of different wastes.

Dynamics of microbial communities related to biochemical parameters during vermicomposting and maturation of agroindustrial lignocellulose wastes.

Scarce information is available on the changes in abundance of microbial taxa during vermicomposting. Quantitative PCR and DGGE analysis were used to monitor variations in the microbial structure, relative abundance of four bacterial classes and fungi over the vermicomposting and maturation period of wet olive cake (O) and vine shoots (W). Multivariate correlation analysis between microbial structure and abundance, earthworm biomass and enzyme activities revealed similar and divergent interactions in both processes. Although Eisenia fetida development was different, significant correlations were found with ß-glucosidase activity and with bacterial and fungal structure. In the vermicomposting period of O and W, a decline was found in bacteria (94% and 77%), fungi (93% and 94%), and Gammaproteobacteria (56% and 71%) but an increase in Betaproteobacteria and Actinobacteria (62-79%). Alphaproteobacteria increased only in O (26%). Despite the different initial lignocellulose wastes, the mature vermicomposts were similar in microbial and biochemical properties.