Rainforest transformation reallocates energy from green to brown food webs.

Terrestrial animal biodiversity is increasingly being lost because of land-use change1,2. However, functional and energetic consequences aboveground and belowground and across trophic levels in megadiverse tropical ecosystems remain largely unknown. To fill this gap, we assessed changes in energy fluxes across 'green' aboveground (canopy arthropods and birds) and 'brown' belowground (soil arthropods and earthworms) animal food webs in tropical rainforests and plantations in Sumatra, Indonesia. Our results showed that most of the energy in rainforests is channelled to the belowground animal food web. Oil palm and rubber plantations had similar or, in the case of rubber agroforest, higher total animal energy fluxes compared to rainforest but the key energetic nodes were distinctly different: in rainforest more than 90% of the total animal energy flux was channelled by arthropods in soil and canopy, whereas in plantations more than 50% of the energy was allocated to annelids (earthworms). Land-use change led to a consistent decline in multitrophic energy flux aboveground, whereas belowground food webs responded with reduced energy flux to higher trophic levels, down to -90%, and with shifts from slow (fungal) to fast (bacterial) energy channels and from faeces production towards consumption of soil organic matter. This coincides with previously reported soil carbon stock depletion3. Here we show that well-documented animal biodiversity declines with tropical land-use change4-6 are associated with vast energetic and functional restructuring in food webs across aboveground and belowground ecosystem compartments.

A review on the fate of micro and nano plastics (MNPs) and their implication in regulating nutrient cycling in constructed wetland systems.

This review discusses the micro-nano plastics (MNPs) and their interaction with physical, chemical and biological processes in a constructed wetland (CW) system that is typically used as a nature-based tertiary wastewater treatment for municipal as well as industrial applications. Individual components of the CW system such as substrate, microorganisms and plants were considered to assess how MNPs influence the CW processes. One of the main functions of a CW system is removal of nutrients like nitrogen (N) and phosphorus (P) and here we highlight the pathways through which the MNPs influence CW's efficacy of nutrient removal. The presence of morphologically (size and shape) and chemically different MNPs influence the growth rate of microorganisms important in N and P cycling, invertebrates, decomposers, and the plants which affect the overall efficiency of a CW treatment system. Certain plant species take up the MNPs, and some toxicity has been observed. This review focuses on two significant aspects: (1) the presence of MNPs in a significant concentration affects the efficiency of N and P removal, and (2) the removal of MNPs. Because MNPs reduce the enzyme activities in abundance and overproduction of ROS oxidizes the enzyme active sites, resulting in the depletion of proteins, ultimately inhibiting nitrogen and phosphorus removal within the substrate layer. The review found that the majority of the studies used sand-activated carbon (SAC), granular-activated carbon (GAC), rice straw, granular limestone, and calcium carbonate, as a substrate for CW treatment systems. Common plant species used in the CW include Phragmites, Arabidopsis thaliana, Lepidium sativum, Thalia dealbata, and Canna indica, which were also found to be dominant in the uptake of the MNPs in the CWs. The MNPs were found to affect earthworms such as Eisenia fetida, Caenorhabditis elegans, and, Enchytraeus crypticus, whereas Metaphire vulgaris were found unaffected. Though various mechanisms take place during the removal process, adsorption and uptake mechanism effectively emphasize the removal of MNPs and nitrogen and phosphorus in CW. The MNPs characteristics (type, size, and concentration) play a crucial role in the removal efficiency of nano-plastics (NPs) and micro-plastics (MPs). The enhanced removal efficiency of NPs compared to MPs can be attributed to their smaller size, resulting in a faster reaction rate. However, NPs dose variation showed fluctuating removal efficiency, whereas MPs dose increment reduces removal efficiency. MP and NPs dose variation also affected toxicity to plants and earthworms as observed from data. Understanding the fate and removal of microplastics in wetland systems will help determine the reuse potential of wastewater and restrict the release of microplastics. This study provides information on various aspects and highlights future gaps and needs for MNP fate study in CW systems.

Alleviation of endothelial dysfunction of Pheretima guillemi (Michaelsen)-derived protein DPf3 in ponatinib-induced thrombotic zebrafish and mechanisms explored through ox-LDL-induced HUVECs and TMT-based proteomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Thrombus generation is one of the leading causes of death in human, and vascular endothelial dysfunction is a major contributor to thrombosis. Pheretima guillemi (Michaelsen), a traditional medicinal animal known as "Dilong", has been utilized to cure thrombotic disorders for many years. DPf3, a group of functional proteins extracted from P. guillemi, has been characterized and identified to possess antithrombotic bioactivity via in vitro and ex vivo experiments. AIM OF THE STUDY: This study is aimed to investigate the vascular-protection activity and related mechanism of antithrombotic protein DPf3 purified from Pheretima guillelmi systematically. MATERIALS AND METHODS: The antithrombotic activity and vascular endothelium protection effect of DPf3 was explored in vivo using ponatinib-induced vascular endothelial injury zebrafish thrombus model. Then, (hi) ox-LDL-induced HUVECs was applied to investigate the protection mechanism of DPf3 against the injury of vascular endothelium. In addition, TMT-based proteomics analysis was used to study the biomarkers, biological processes and signal pathways involved in the antithrombotic and vascular protective effects of DPf3 holistically. RESULTS: DPf3 exerted robust in vivo antithrombosis and vascular endothelial protection ability. DPf3 was identified to prevent HUVECs from damage by reducing ROS production, and to reduce monocyte adhesion by decreasing the protein content of adhesion factor VCAM 1. DPf3 was also observed to weaken the migration ability of injured cells and inhibit abnormal angiogenesis. The mechanism of DPf3's antithrombotic and vascular protective activity was mainly related to the regulation of lipid metabolism, energy metabolism, complement and coagulation system, ECM receptor interaction, MAPK signal pathway, etc. CONCLUSIONS: This study demonstrates that DPf3 has strong antithrombotic and endothelial protective effects. The endothelial protective ability and related mechanisms of DPf3 provide a scientific reference for the traditional use of earthworms in the treatment of thrombosis.

The Protective Effect of Lithium Against Rotenone may be Evolutionarily Conserved: Evidence from Eisenia fetida, a Primitive Animal with a Ganglionic Brain.

Chronic exposure to stress is a non-adaptive situation that is associated with mitochondrial dysfunction and the accumulation of reactive oxygen species (ROS), especially superoxide anion (SA). This accumulation of ROS produces damage-associated molecular patterns (DAMPs), which activate chronic inflammatory states and behavioral changes found in several mood disorders. In a previous study, we observed that an imbalance of SA triggered by rotenone (Ro) exposure caused evolutionarily conserved oxi-inflammatory disturbances and behavioral changes in Eisenia fetida earthworms. These results supported our hypothesis that SA imbalance triggered by Ro exposure could be attenuated by lithium carbonate (LC), which has anti-inflammatory properties. The initial protocol exposed earthworms to Ro (30 nM) and four different LC concentrations. LC at a concentration of 12.85 mg/L decreased SA and nitric oxide (NO) levels and was chosen to perform complementary assays: (1) neuromuscular damage evaluated by optical and scanning electron microscopy (SEM), (2) innate immune inefficiency by analysis of Eisenia spp. extracellular neutrophil traps (eNETs), and (3) behavioral changes. Gene expression was also evaluated involving mitochondrial (COII, ND1), inflammatory (EaTLR, AMP), and neuronal transmission (nAchR α5). LC attenuated the high melanized deposits in the circular musculature, fiber disarrangement, destruction of secretory glands, immune inefficiency, and impulsive behavior pattern triggered by Ro exposure. However, the effects of LC and Ro on gene expression were more heterogeneous. In summary, SA imbalance, potentially associated with mitochondrial dysfunction, appears to be an evolutionary component triggering oxidative, inflammatory, and behavioral changes observed in psychiatric disorders that are inhibited by LC exposure.

Bioremediation of polycyclic aromatic hydrocarbons in crude oil by bacterial consortium in soil amended with Eisenia fetida and rhamnolipid.

The present study investigated the concerted effort of Eisenia fetida and rhamnolipid JBR-425 in combination with a five-member bacterial consortium exhibiting elevated degradation levels of low and high molecular weight polycyclic aromatic hydrocarbons (PAH) from soil contaminated with Digboi crude oil. Application of bacterial consortium (G2) degraded 30-89% of selected PAH from the artificial soil after a 45-day post-exposure, in which chrysene showed the highest level of degradation with 89% and benzo(a)pyrene is the lowest with 30%, respectively. Moreover, an acute exposure study observed that earthworm biomass decreased, and mortality rates increased with increasing crude oil concentrations (0.25 to 2%). Earthworms with a 100% survival rate at 1% crude oil exposure suggest the tolerance potential and its mutual involvement in the bioremediation of crude oil with selected bacterial consortia. Bacterial consortium assisted with E. fetida (G3) showed 98% chrysene degradation with a slight change in benzo(a)pyrene degradation (35%) in crude oil spiked soil. Besides, the most dominant PAH in crude oil found in the current work, fluoranthene, undergoes 93% and 70% degradation in G3 and G5 groups, respectively. However, rhamnolipid JBR-425 coupled with the bacterial consortium (G5) has resulted in 97% degradation of chrysene and 33% for benzo(a)pyrene. Overall, bacterial consortium assisted with earthworm group has shown better degradation of selected PAH than bacterial consortium with biosurfactant. Catalase (CAT), glutathione reductase (GST) activity and MDA content was found to be reduced in earthworms after sub-lethal exposure, suggesting oxidative stress prevalence via reactive oxygen species (ROS). Hence, the findings of the present work suggest that the application of a bacterial consortium, along with earthworm E. fetida, has huge potential for field restoration of contaminated soil with PAH and ecosystem sustainability.

Toxicity of methylparaben and its chlorinated derivatives to Allium cepa L. and Eisenia fetida Sav.

Methylparaben, chloro-methylparaben, and dichloro-methylparaben were evaluated in Allium cepa at 5, 10, 50, and 100 µg/L and in Eisenia fetida at 10 and 100 µg/L. In A. cepa roots, 100 µg/L methylparaben and 50 and 100 µg/L chlorinated methylparabens reduced cell proliferation, caused cellular changes, and reduced cell viability in meristems, which caused a reduction in root growth. Furthermore, they caused drastic inhibition of catalase, ascorbate peroxidase, and superoxide dismutase; activated guaiacol peroxidase and promoted lipid peroxidation in meristematic root cells. In earthworms, after 14 days exposure to the three compounds, there were no deaths, and catalase, ascorbate peroxidase, and superoxide dismutase were not inhibited. However, guaiacol peroxidase activity and lipid peroxidation were observed in animals exposed to dichloro-methylparaben. Soils with dichloro-methylparaben also caused the escape of earthworms. It is inferred that the recurrent contamination of soils with these methylparabens, with emphasis on chlorinated derivatives, can negatively impact different species that depend directly or indirectly on soil to survive.

Uptake, distribution, and elimination of selenite in earthworm Eisenia fetida at sublethal concentrations based on toxicokinetic model.

Natural and anthropogenic causes have promoted the rapid increase in environmental selenium (Se) levels, and the complex Se metabolism and dynamic in organisms make it challenging to evaluate the toxicity and ecological risks. In this study, the kinetics of selenite in earthworm Eisenia fetida were investigated based on toxicokinetic (TK) model (uptake-elimination phases: 14-14 days). The results showed the highest sub-tissue Se concentrations in pre-clitellum (PC), post-clitellum (PoC) parts, and total earthworms were 95.71, 70.40, and 79.94 mg/kg, respectively, which indicates the distinctive Se uptake capacities of E. fetida. Se kinetic rates in PCs were faster than that of the total E. fetida for both uptake (Kus = 0.30-0.80 mg/kg/day) and elimination phases (Kee = 0.024-0.056 mg/kg/day). Longer half-life times (LT1/2) were observed in the total earthworms (17.85-47.15 d) than PCs (12.28-29.22 d), while non-significant difference was found for the kinetic Se bioaccumulation factor (BAFk) in PC and total earthworms (12-19), which demonstrates that Se can be efficiently bioaccumulated and eliminated in earthworm PC part. Besides, the significant increase Se concentration in PoC with rapid elimination in PC also illustrates that earthworms can alleviate the Se stress by the transformation strategy of Se from the head to tail tissues. In conclusion, the investigation of Se kinetic accumulation and elimination characteristics in this study is helpful for understanding the metabolism and detoxification processes of Se in earthworms, and also providing a theoretical basis for further Se risk assessment using TK model.

Effects of hydrolyzed polymaleic anhydride addition combined with vermicomposting on maturity and bacterial diversity in the final vermicompost from the biochemical residue of kitchen waste.

A large amount of kitchen waste is produced all over the world. Biochemical disposal is an effective method for the reduction and safe utilization of kitchen waste. However, high salinity, low maturity and poor biocompatibility were encountered when utilizing the biochemical residue of kitchen waste (BRKW) as a kind of soil amendment. To reduce the high salinity, accelerate the maturity and improve the biocompatibility in the BRKW, this study used the BRKW as the main feedstock for earthworms after hydrolyzed polymaleic anhydride (HPMA) was added and focused on revealing the effect of HPMA addition combined with the vermicomposting process on the growth of earthworms and on the basic physicochemical properties and the microbial diversity of the derived vermicompost. The results showed that HPMA addition can promote earthworm growth and reproduction. The pH, electric conductivity, organic matter content, C/N and NH4+-N/NO3--N were decreased in the final vermicompost, while total nitrogen, total phosphorus and total potassium contents, and the seed germination index were increased. Scanning electron microscopy analysis showed that there was more disintegration in the final vermicompost. Meanwhile, adding the HPMA also helped to decrease the total number of fungi while increasing the populations of nitrogen-fixing bacteria, phosphorus-solubilizing bacteria and potassium-solubilizing bacteria as well as amount of total bacteria and actinomycetes. The vermicomposting process increased the bacterial phyla that promote the degradation of OM, such as Actinobacteria, Firmicutes and Acidobacteria, decreased the pathogenic Enterobacter and increased the bacterial genera that promote the maturity and quality, such as Cellvibrio and Pseudomonas. Thus, HPMA addition combined with vermicomposting can promote the growth of beneficial bacteria that promote the degradation of lignocelluloses and accelerate maturity while inhibiting some potential bacterial pathogens, which helps guarantee the safety of vermicomposting products from BRKW. Hence, employing HPMA to promote BRKW vermicomposting can possibly reduce salt content and improve the maturity and biocompatibility of the final vermicompost. This approach may help realize the safe utilization of BRKW and further promote the biochemical disposal of kitchen waste.

Bioremediation of hydrocarbon contaminated soil using local organic materials and earthworms.

Bioremediation technologies have demonstrated significant success on biological quality recovery of hydrocarbon contaminated soils, employing techniques among which composting and vermiremediation stand out. The aim of this study was to evaluate the efficiency of these processes to remediate diesel-contaminated soil, employing local organic materials and earthworms. During the initial composting stage (75 days), the substrate was made up using contaminated soil, lombricompost, rice hulls and wheat stubbles (60:20:15:5% w/w). Diesel concentration in the contaminated substrate was about 5 g kg-1, equivalent to a Total Petroleum Hidrocarbons (TPH) experimental concentration of 3425 ± 50 mg kg-1. During the later vermiremediation stage (60 days), the earthworm species Eisenia fetida and Amynthas morrisi were evaluated for their hydrocarbon degradation capacity. Physicochemical and biological assays were measured at different times of each stage and ecotoxicity assays were performed at the end of the experiments. TPH concentration reduced 10.91% after composting and from 45.2 to 60.81% in the different treatments after vermiremediation. Compared with TPH degradation in the treatment without earthworms (16.05%), results indicate that earthworms, along with indigenous microorganisms, accelerate the remediation process. Vermiremediation treatments did not present phytotoxicity and reflected high substrate maturity values (>80% Germination Index) although toxic effects were observed due to E. fetida and A morrisi exposure to diesel. Vermiremediation was an efficient technology for the recovery of substrate biological quality after diesel contamination in a short period. The addition of organic materials and suitable food sources aided earthworm subsistence, promoted the decontamination process and improved the substrate quality for future productive applications.

A Strategy for Rapid Discovery of Marker Peptides Associated with Fibrinolytic Efficacy of Pheretima aspergillum Based on Bioinformatics Combined with Parallel Reaction Monitoring.

Quality control of animal-derived traditional Chinese medicines has improved dramatically as proteomics research advanced in the past few decades. However, it remains challenging to identify quality attributes with routine proteomics approaches since protein with fibrinolytic activity is rarely reported in pheretima, a typical animal-derived traditional medicine. A novel strategy based on bioinformatics combined with parallel reaction monitoring (PRM) was developed here to rapidly discover the marker peptides associated with a fibrinolytic effect. Potential marker peptides were found by lumbrokinase sequences' alignment and in silico digestion. The fibrinogen zymography was used to visually identify fibrinolytic proteins in pheretima. As a result, it was found that the fibrinolytic activity varied among different portions of pheretima. Fibrinolytic proteins were distributed regionally in the anterior and anterior-mid portion and there was no significant fibrinogenolytic activity observed in the mid-posterior and posterior portion. Finally, PRM experiments were deployed to validate and quantify selected marker peptides and a total of 11 peptides were identified as marker peptides, which could be potentially used in quality control of pheretima. This strategy provides a robust workflow to benefit the quality control of other animal-derived traditional medicines.

Commonwealth of Soil Health: How Do Earthworms Modify the Soil Microbial Responses to CeO2 Nanoparticles?

Soil ecotoxicological assays on nanoparticles (NPs) have mainly investigated single components (e.g., plants, fauna, and microbes) within the ecosystem, neglecting possible effects resulting from the disturbance of the interactions between these components. Here, we investigated soil microbial responses to CeO2 NPs in the presence and absence of earthworms from the perspectives of microbial functions (i.e., enzyme activities), the community structure, and soil metabolite profiles. Exposure to CeO2 NPs (50, 500 mg/kg) alone decreased the activities of enzymes (i.e., acid protease and acid phosphatase) participating in soil N and P cycles, while the presence of earthworms ameliorated these inhibitory effects. After the CeO2 NP exposure, the earthworms significantly altered the relative abundance of some microbes associated with the soil N and P cycles (Flavobacterium, Pedobacter, Streptomyces, Bacillus, Bacteroidota, Actinobacteria, and Firmicutes). This was consistent with the pattern found in the significantly changed metabolites which were also involved in the microbial N and P metabolism. Both CeO2 NPs and earthworms changed the soil bacterial community and soil metabolite profiles. Larger alterations of soil bacteria and metabolites were found under CeO2 NP exposure with earthworms. Overall, our study indicates that the top-down control of earthworms can drastically modify the microbial responses to CeO2 NPs from all studied biological aspects. This clearly shows the importance of the holistic consideration of all soil ecological components to assess the environmental risks of NPs to soil health.

Vermicomposting of textile mill sludge employing Eisenia fetida: Role of cow dung and tea waste amendments.

Vermicomposting of textile mill sludge (TMS) with cow dung (CD) and tea waste (TW) as amendments was done in seven different combinations using Eisenia fetida for 90 days. Results revealed that pH decreased from 7.68-8.63 to 7.09-7.59. TOC content and C/N ratio reductions were in range of 15.71-20.08% and 39.33-50.05%, respectively (P < 0.05). The macronutrients in the form of TN, TP, and TK increased 0.38-0.64, 1.07-2.27, and 0.56-1.98 times respectively after end of bioconversion process (P < 0.05), among increases in ash content and EC. The biomass and cocoon production of E. fetida increased significantly (P < 0.05), while high mortality rate of earthworms was observed in treatments containing 50% or more TMS content. The bacterial population beneficial for degradation of organic matter increased significantly over initial substrates (0th day) (P < 0.05). Increased humification index in end-product indicated better maturity of vermicompost as observed in treatments containing higher proportions of amendments. The addition of amendments favored earthworm activity which significantly decreased the heavy metal concentration (Fe, Cu, Pb, Zn) in the end-product. The study concluded that sustainable utilization of TMS could be achieved for cleaner and enriched vermicompost production by addition of amendments CD and TW in proportions of 50% and above.

A rapid and simple signature peptides-based method for species authentication of three main commercial Pheretima.

Pheretima with various activities is a commonly used animal-derived traditional medicine in Asia countries. However, almost half of them are non-pharmacopoeia species in the market due to the similar morphological characteristics between medicinal and non-medicinal species. This study aims to establish an effective method based on signature peptides for species authentication of three main commercial Pheretima, including two major Pheretima species (Amynthas aspergillum, Metaphire vulgaris) and one main adulteration (Metaphire magna). Firstly, the species of 52 batches of commercial Pheretima were authenticated based on DNA barcodes. Secondly, proteomic analysis was performed for protein characterization of three main commercial Pheretima. Furthermore, their signature peptides were screened and validated using ultra-high performance liquid chromatography coupled with mass spectrometry (UPLC-MS/MS) in multiple reaction monitoring (MRM) mode. Moreover, a simplified sample processing method was developed. Finally, large quantities of commercial Pheretima samples were analyzed for further verifying the feasibility of the signature peptides-based method. The result showed that the established method had a great application potential for authenticity identification of commercial Pheretima. SIGNIFICANCE: The authenticity assessment of medicinal materials is a main issue in the quality control process as deceptive practices could imply severe health risks. In this study, a rapid and simple method based on signature peptides was established for species authentication of three main commercial Pheretima, which can be an effective alternative to complex DNA barcoding and difficult morphological identification, and provided a reference for improvement of Pheretima quality standards.

Toxic mechanism on phenanthrene-induced cytotoxicity, oxidative stress and activity changes of superoxide dismutase and catalase in earthworm (Eisenia foetida): A combined molecular and cellular study.

Phenanthrene (PHE) is an important organic compound, which is widespread in the soil environment and exhibits potential threats to soil organisms. Toxic effects of PHE to earthworms have been extensively studied, but toxic mechanisms on PHE-induced cytotoxicity and oxidative stress at the molecular and cellular levels have not been reported yet. Therefore, we explored the cytotoxicity and oxidative stress caused by PHE in earthworm coelomocytes and the interaction mechanism between PHE and the major antioxidant enzymes SOD/CAT. It was shown that high-dose PHE exposure induced the intracellular reactive oxygen species (ROS) generation, mediated lipid peroxidation, reduced total antioxidant capacity (T-AOC) in coelomocytes, and triggered oxidative stress, thus resulted in a strong cytotoxicity at higher concentrations (0.6-1.0 mg/L). The intracellular SOD/CAT activity in cells after PHE exposure were congruent with that in molecular levels, which the activity of SOD enhanced and CAT inhibited. Spectroscopic studies showed the SOD/CAT protein skeleton and secondary structure, as well as the micro-environment of aromatic amino acids were changed after PHE binding. Molecular docking indicated PHE preferentially docked to the surface of SOD. However, the key residues Tyr 357, His 74, and Asn 147 for activity were in the binding pocket, indicating PHE more likely to dock to the active center of CAT. In addition, H-bonding and hydrophobic force were the primary driving force in the binding interaction between PHE and SOD/CAT. This study indicates that PHE can induce cytotoxicity and oxidative damage to coelomocytes and unearthes the potential effects of PHE on earthworms.

Marker peptide screening and species-specific authentication of Pheretima using proteomics.

Pheretima is a common and valuable animal-derived medication used in traditional Chinese medicine. There are four species of Pheretima specified in the Chinese Pharmacopoeia (2015 edition), i.e. Pheretima aspergillum, P. vulgaris, P. guillelmi, and P. pectinifera. A recent report revealed ~ 55% of Pheretima in the commercial marketplace may be adulterated by other species, contrary to the Pharmacopoeia standard. The safety, efficacy, and authenticity of Pheretima is an important issue. Currently, the availability of specific quality-markers for the various species and effective identification methods are still limited. In this study, label-free quantification proteomics of species from Pheretima and Amynthas was carried out using nanoscale liquid chromatography coupled to tandem mass spectrometry (nano LC-MS/MS), and marker peptides were identified based on their ion intensities using multivariate data analysis (principal component analysis and supervised partial least-squares discriminant analysis). A total of 48,476 peptides with high confidence corresponding to 13,397 proteins were identified from all samples. The marker peptides were validated by comparison with synthetic peptide reference standards using LC-MS/MS operating in a multiple-reaction monitoring mode. A multiple-peptide identification strategy was proposed for the authentication of Pheretima and subsequently applied to samples obtained from retail outlets in various regions of China. The results showed that eight out of the 15 samples tested were deemed authentic Pheretima.

Vermicomposting of Organic Waste with Eisenia fetida Increases the Content of Exchangeable Nutrients in Soil.

BACKGROUND AND OBJECTIVE: Vermicomposting is a green technique used to produce organic compost from organic waste with the aid of specific earthworm species. The resulting compost is rich in nutrients that can improve plant health and fertility. This study was conducted to produce organic compost using a developed vermicomposting technique. And that is to enhance and increase the exchangeable nutritional content in the soil for utilization in sustainable agriculture. MATERIALS AND METHODS: The experiment was carried out with Eisenia fetida worms in a treatment technique using sugarcane straw, remnants of garden tree leaves, kitchen wastes and cow manure. The physiochemical parameters, namely temperature, moisture, acidity, electrical conductivity and 10 different nutrients, were assessed in vermicompost samples and garden soil sample (without earthworms). RESULTS: The results revealed higher N, C and P content in the vermicompost than in the garden soil sample. CONCLUSION: This study confirms that vermicompost has a high nutrient value and can be considered a promising method for safely disposing organic waste, improving crop production and achieving long-term food security.

Ecotoxicological effects of petroleum-contaminated soil on the earthworm Eisenia fetida.

Petroleum is an important industrial raw material that enters the soil during production and use and is harmful to soil organisms. To evaluate the toxicity of petroleum-contaminated soil, earthworms (Eisenia fetida) were used as model organisms for soil ecotoxicity studies. We found that earthworm weight and cocoon production decreased significantly after exposure to petroleum-contaminated soil. In addition, soil contaminated with high concentrations of petroleum can cause damage to the DNA within earthworm seminal vesicles. Superoxide dismutase (SOD), catalase, and peroxidase activities were significantly inhibited when earthworms were exposed to petroleum-contaminated soil, indicating that oxidative stress was induced by petroleum pollutants. The mRNA levels of annetocin precursor, a reproduction-related gene, was significantly inhibited after petroleum exposure. The mRNA levels of translationally controlled tumour protein (TCTP) and SOD exhibited a concentration-dependent relationship, and their relative expression increased with petroleum concentration.

Transcriptomic-proteomics-anticoagulant bioactivity integrated study of Pheretima guillemi.

ETHNOPHARMACOLOGICAL RELEVANCE: Earthworms, a type of animal drugs from traditional Chinese medicine, have been used to treat coagulation for many years with less adverse effects and similar anticoagulant effects compared to the commonly used anticoagulants. There are four species of earthworms recorded in Chinese Pharmacopoeia, while few of them were studied and deficient information were involved in the NCBI and UniProt earthworm protein database. We have adopted a transcriptomic-proteomics-anticoagulant bioactivity integrated approach to investigate a seldom-studied Chinese Pharmacopoeia recorded species, Pheretima guillelmi. AIM OF THE STUDY: In the present study, we aimed to reveal the anticoagulant bioactivity of Pheretima guillelmi, and identify its functional proteins via LC-MS/MS-transcriptome cross identification. METHODS AND RESULTS: With the aid of fibrinogen-thrombin time assay, Pheretima guillelmi was found to possess strong anticoagulant activity, and the bioactivity was quite stable under 30-50 °C and near-neutral conditions. A comprehensive non-reference transcriptome assembly of P. guillelmi was first established to supplement the currently inadequate earthworm protein database and to illustrate the active proteins. Illumina RNA sequencing generated 25,931,175 of clean reads with over 97% high-quality clean reads (Q20) and assembled an average of 133,228 of transcript and 106,717 of unigenes. A total of 11,259 coding sequences were predicted via ESTScan (3.0.3). The P. guillelmi unigenes were searched and annotated against public database. The bioactive proteins in P. guillelmi were with broad distribution of molecular weight. With bottom-up proteomics analysis, ten proteins were identified against UniProt and NCBI earthworm database; and 31 proteins with high-confidence were matched against transcriptomic established P. guillelmi database. CONCLUSION: This study illuminated the therapeutic potency of P. guillelmi for antithrombus and provide a new strategy to investigate animal drugs of Chinese materia medica.

Bio-Optimization of Chemical Parameters and Earthworm Biomass for Efficient Vermicomposting of Different Palm Oil Mill Waste Mixtures.

The present study reports mathematical modelling of palm oil mill effluent and palm-pressed fiber mixtures (0% to 100%) during vermicomposting process. The effects of different mixtures with respect to pH, C:N ratio and earthworms have been optimized using the modelling parameters. The results of analysis of variance have established effect of different mixtures of palm oil mill effluent plus palm press fiber and time, under selected physicochemical responses (pH, C:N ratio and earthworm numbers). Among all mixtures, 60% mixture was achieved optimal growth at pH 7.1 using 16.29 C:N ratio in 15 days of vermicomposting. The relationship between responses, time and different palm oil mill waste mixtures have been summarized in terms of regression models. The obtained results of mathematical modeling suggest that these findings have potential to serve a platform for further studies in terms of kinetic behavior and degradation of the biowastes via vermicomposting.

An ecological microsystem to treat waste oil contaminated soil: Using phytoremediation assisted by fungi and local compost, on a mixed-contaminant site, in a cold climate.

As a result of anthropization and industrialization, northern remote communities face issues of soil contamination by mixtures of organic and inorganic contaminants. Soil bioremediation in cold environments is particularly challenging because of slower degradation rates, slower production of biomass for phytoextraction of trace elements (TEs), and remoteness, which can complicate logistics and inflate costs. This study evaluated a decontamination approach integrating indigenous willows, fungi and compost in a northern community. The site was a waste oil pit and its soil was initially contaminated with petroleum hydrocarbons (PHC) exceeding 200 g kg-1 and TEs including As, Cd, Co, Cr, Cu, Pb and Zn. In under five years, 65 and 75% of PHC (C6-C50 and >C50) were degraded, compared to 27 and 13% for the untreated control soil. We found contrasting TE translocation patterns to the aboveground biomass for the willow species used (Salix planifolia and Salix alaxensis), as well as distinctive rooting strategies. Hazard quotients were calculated to assess the risk plant material could pose to local wildlife. The highest TE concentration measured was Zn in S. planifolia, which exceeded Canadian soil guidelines. Results indicate toxicity risks to animals linked to TEs in Salix spp. leaves is generally unlikely. The fungus Trametes versicolor inoculated into the soil did not fruit, however fruiting bodies of Psathyrella sp. were observed consistently (four out of five years). Biological tests indicated that in five growing seasons soil toxicity significantly decreased compared to the untreated soil used as control. This was demonstrated by vegetation cover (137 vs 11% cover), toxicity assays on earthworms (Eisenia andrei) (0 vs 33% mortality) and barley seed germination (Hordeum vulgare) (86 vs 62% germination). The proposed decontamination approach, without the use of synthetic fertilizers, is promising for the PHC remediation of mixed-contaminants on cold climate sites.