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Abstract With the increasing demand for organic food production, the earthworm is used as a soil nutrient facilitator. The present study was conducted to assess the effect of epigeic earthworms Perionyx excavatus and Eisenia. fetida on soil nutrients and the consequent improvement of biomass productivity and yield of Capsicum chinense Jacq and Zea mays L. The experiment was conducted in 5 L and 15 L capacity plastic pots for C. chinense and Z. mays with 150 g and 300 g of half-decomposed cow dung, respectively. It was observed that the weekly harvest rate of ripened chili was 17.59 g, 13.91 g, and 9.24 g in P. excavatus, control, and E. fetida pot showing 26.49% higher in P. excavatus. Also, the total kernel count per corn was significantly different (F (2, 9) = 37.78, p

With the increasing demand for organic food production, the earthworm is used as a soil nutrient facilitator. The present study was conducted to assess the effect of epigeic earthworms Perionyx excavatus and Eisenia. fetida on soil nutrients and the consequent improvement of biomass productivity and yield of Capsicum chinense Jacq and Zea mays L. The experiment was conducted in 5 L and 15 L capacity plastic pots for C. chinense and Z. mays with 150 g and 300 g of half-decomposed cow dung, respectively. It was observed that the weekly harvest rate of ripened chili was 17.59 g, 13.91 g, and 9.24 g in P. excavatus, control, and E. fetida pot showing 26.49% higher in P. excavatus. Also, the total kernel count per corn was significantly different (F(2, 9) = 37.78, p < 0.05), with the highest kernel present in P. excavatus(333.5 ± 13.5), followed by E. fetida(261.5 ± 16.5) and control (235 ± 22). The impact of P. excavatus was more perceptible in C. chinense, indicated by higher leaf biomass (69.16%), root length (30.14%), and fruit harvest (71.03%). However, the effect of E. fetida was noticed more in Z. mays (stem length, 19.24%, stem biomass, 14.39%, root biomass, 20.9%, kernel count, 41.91%, and kernel weight, 95.07%). Enhanced plant productivity was also supported by an increasing soil nutrient turnover in organic carbon (OC) (25.76% and 23.4%), Phosphorus (P) (31.03% and 25.67%), and Potassium (K) (41.67% and 12.26) in P. excavatus and E. fetida worked soil respectively. The findings indicate that earthworms have a notable impact on plant biomass productivity by promoting the mineralization of soil nutrients and imply on possibility of organic cultivation of seasonal vegetables without using synthetic fertilizers. [ABSTRACT FROM AUTHOR]

Abstract Triphenyl phosphate (TPHP) is one of the most highly utilized organophosphorus flame retardants, and has been frequently detected in various environmental matrices, including soil. So far, limited information is known regarding the potential toxicity of TPHP to the earthworm-soil ecosystem. We investigated the metabolism of TPHP and the perturbation of the endogenous metabolome in the earthworm, Perionyx excavatus, using gas chromatography mass spectrometry (GC-MS) and liquid chromatography quadrupole time-of-flight (LC-QTOF)-based untargeted metabolomics approach after acute exposure to TPHP for one and two days through a filter paper contact test, as well as after chronic exposure for 28 days in a soil microcosm experiment. TPHP showed low bioaccumulation potential in the earthworm-soil ecosystem at concentrations of 10 mg/kg and 50 mg/kg. Identified phase I metabolites include diphenyl phosphate, mono-hydroxylated and di-hydroxylated TPHP. Two groups of phase II metabolites, thiol conjugates (including mercaptolactic acid, cysteine, cysteinylglycine, and mercaptoethanol conjugates) and glucoside conjugates (including glucoside, glucoside-phosphate, and C14H19O10P conjugates), were putatively identified. Only acute TPHP exposure caused significant perturbations of the endogenous metabolome in earthworms, featuring fluctuations in amino acids, glucose, inosine and phospholipids. These results reveal novel phase II metabolism and toxicity of TPHP in P. excavatus.

Abstract Fetal bovine serum (FBS) plays a pivotal role in animal cell culture. Due to ethical and scientific issues, searching for an alternative, comprising the three R’s (Refinement, Reduction and Replacement) gained global attention. In this context, we have identified the heat inactivated coelomic fluid (HI-CF) of the earthworm, Perionyx excavatus as a potential alternative for FBS. Briefly, we formulated HI-CF (f-HICF) containing serum free medium which can aid the growth, attachment, and proliferation of adherent cells, similar to FBS. In this study, we investigated the biochemical characterization, sterility, stability, formulation, and functional analysis of HI-CF as a supplement in culturing animal cells. Notably, vitamins, micronutrients, proteins, lipids, and trace elements are identified and compared with FBS for effective normalization of the serum free media. HI-CF is tested to be devoid of endotoxin and mycoplasma contamination thus can qualify the cell culture grade. The f-HICF serum free media was prepared, optimised, and tested with A549, HeLa, 3T3, Vero and C2C12 cell lines. Our results conclude that f-HICF is a potential alternative to FBS, in accordance with ethical concern; compliance with 3R's; lack of unintended antibody interactions; presence of macro and micronutrients; simple extraction; cost-effectiveness and availability.

Triphenyl phosphate (TPHP) is one of the most highly utilized organophosphorus flame retardants, and has been frequently detected in various environmental matrices, including soil. So far, limited information is known regarding the potential toxicity of TPHP to the earthworm-soil ecosystem. We investigated the metabolism of TPHP and the perturbation of the endogenous metabolome in the earthworm, Perionyx excavatus, using gas chromatography mass spectrometry (GC-MS) and liquid chromatography quadrupole time-of-flight (LC-QTOF)-based untargeted metabolomics approach after acute exposure to TPHP for one and two days through a filter paper contact test, as well as after chronic exposure for 28 days in a soil microcosm experiment. TPHP showed low bioaccumulation potential in the earthworm-soil ecosystem at concentrations of 10 mg/kg and 50 mg/kg. Identified phase I metabolites include diphenyl phosphate, mono-hydroxylated and di-hydroxylated TPHP. Two groups of phase II metabolites, thiol conjugates (including mercaptolactic acid, cysteine, cysteinylglycine, and mercaptoethanol conjugates) and glucoside conjugates (including glucoside, glucoside-phosphate, and C14H19O10P conjugates), were putatively identified. Only acute TPHP exposure caused significant perturbations of the endogenous metabolome in earthworms, featuring fluctuations in amino acids, glucose, inosine and phospholipids. These results reveal novel phase II metabolism and toxicity of TPHP in P. excavatus. [ABSTRACT FROM AUTHOR]

Fetal bovine serum (FBS) plays a pivotal role in animal cell culture. Due to ethical and scientific issues, searching for an alternative, comprising the three R's (Refinement, Reduction and Replacement) gained global attention. In this context, we have identified the heat inactivated coelomic fluid (HI-CF) of the earthworm, Perionyx excavatus as a potential alternative for FBS. Briefly, we formulated HI-CF (f-HICF) containing serum free medium which can aid the growth, attachment, and proliferation of adherent cells, similar to FBS. In this study, we investigated the biochemical characterization, sterility, stability, formulation, and functional analysis of HI-CF as a supplement in culturing animal cells. Notably, vitamins, micronutrients, proteins, lipids, and trace elements are identified and compared with FBS for effective normalization of the serum free media. HI-CF is tested to be devoid of endotoxin and mycoplasma contamination thus can qualify the cell culture grade. The f-HICF serum free media was prepared, optimised, and tested with A549, HeLa, 3T3, Vero and C2C12 cell lines. Our results conclude that f-HICF is a potential alternative to FBS, in accordance with ethical concern; compliance with 3R's; lack of unintended antibody interactions; presence of macro and micronutrients; simple extraction; cost-effectiveness and availability. [ABSTRACT FROM AUTHOR]

The present studies show the effect of the Venetin-1 protein-polysaccharide complex obtained from the coelomic fluid of the earthworm Dendrobaena veneta on Candida albicans cells. They are a continuation of research on the mechanisms of action, cellular targets, and modes of cell death. After the action of Venetin-1, a reduced survival rate of the yeast cells was noted. The cells were observed to be enlarged compared to the controls and deformed. In addition, an increase in the number of cells with clearly enlarged vacuoles was noted. The detected autophagy process was confirmed using differential interference contrast, fluorescence microscopy, and transmission electron microscopy. Autophagic vesicles were best visible after incubation of fungus cells with the Venetin-1 complex at a concentration of 50 and 100 µg mL−1. The changes in the vacuoles were accompanied by changes in the size of mitochondria, which is probably related to the previously documented oxidative stress. The aggregation properties of Venetin-1 were characterized. Based on the results of the zeta potential at the Venetin-1/KCl interface, the pHiep = 4 point was determined, i.e. the zeta potential becomes positive above pH = 4 and is negative below this value, which may affect the electrostatic interactions with other particles surrounding Venetin-1. [ABSTRACT FROM AUTHOR]

Maintaining genomic stability is inevitable for organism survival and it is challenged by mutagenic agents, which include ultraviolet (UV) radiation. Whenever DNA damage occurs, it is sensed by DNA-repairing proteins and thereby performing the DNA-repair mechanism. Specifically, in response to DNA damage, H2AX is a key protein involved in initiating the DNA-repair processes. In this present study, we investigate the effect of UV-C on earthworm, Perionyx excavatus and analyzed the DNA-damage response. Briefly, we expose the worms to different doses of UV-C and find that worms are highly sensitive to UV-C. As a primary response, earthworms produce coelomic fluid followed by autotomy. However, tissue inflammation followed by death is observed when we expose worm to increased doses of UV-C. In particular, UV-C promotes damages in skin layers and on the contrary, it mediates the chloragogen and epithelial outgrowth in intestinal tissues. Furthermore, UV-C promotes DNA damages followed by upregulation of H2AX on dose-dependent manner. Our finding confirms DNA damage caused by UV-C is directly proportional to the expression of H2AX. In short, we conclude that H2AX is present in the invertebrate earthworm, which plays an evolutionarily conserved role in DNA damage event as like that in higher animals. [ABSTRACT FROM AUTHOR]

In the present research, the effect of a protein-polysaccharide complex Venetin-1 obtained from the coelomic fluid of Dendrobaena veneta earthworm on Candida albicans cells was characterized. The compound destroyed fungal cells without showing cytotoxicity to human skin fibroblasts, which was demonstrated in earlier studies. Since it had an effect on the fungal cell wall and membrane, this complex was compared with the known antifungal antibiotic fluconazole. Both preparations disturbed the division of yeast cells and resulted in the formation of aggregates and chains of unseparated cells, which was illustrated by staining with fluorochromes. Fluorescent staining of the cell wall with Calcofluor white facilitated comparison of the types of aggregates formed after the action of both substances. The analysis performed with the use of Congo red showed that Venetin-1 exposed deeper layers of the cell wall, whereas no such effect was visible after the use of fluconazole. The FTIR analysis confirmed changes in the mannoprotein layer of the cell wall after the application of the Venetin-1 complex. Staining with Rhodamine 123 and the use of flow cytometry allowed comparison of changes in the mitochondria. Significantly elongated mitochondria were observed after the Venetin-1 application, but not after the application of the classic antibiotic. Phase contrast microscopy revealed vacuole enlargement after the Venetin-1 application. The flow cytometry analysis of C. albicans cells treated with Venetin-1 and fluconazole showed that both substances caused a significant decrease in cell viability. [ABSTRACT FROM AUTHOR]

Himalayan ecosystem is characterized by its fragile climate with rich repositories of biodiversity. Waste collection and disposal are becoming increasingly difficult due to topographical variations. Aporrectodea caligenosa, a versatile psychrophillic soil dweller, is a useful biocatalyst with potent bio-augmented capability for waste treatment at low temperatures. Microcosm experiments were conducted to elucidate the comprehensive nature of biogenic nitrogen transformation to NH4+ and NO3− produced by coupling of earthworm-microbes. Higher biogenic recovery of NH4+-N from coprolites of garden soil (47.73 ± 1.16%) and Himalayan goat manure (86.32 ± 0.92%) with an increment of 14.12 and 47.21% respectively over their respective control (without earthworms) with a linear decline beyond 4th week of incubation was reported. NO3–-N recovery progressively sustained in garden soil and goat manure coprolites during entire incubation with highest 81.81 ± 0.45 and 87.20 ± 1.08 µg-N g−1dry weight recorded in 6th and 5th week of incubation respectively and peak increments as 38.58 and 53.71% relative to respective control (without earthworms). Declined NH4+–N in coprolites at low temperature (15.0 ± 2.0 °C) evidenced increased nitrification rates by taking over the process by abundant nitrifying microbes. Steady de-nitrification with progressive incubation on an average was 16.95 ± 0.46 ng-N g−1 per week and 21.08 ± 0.87 ng-N g−1 per week compared to 14.03 ± 0.58 ng-N g−1 per week and 4.50 ± 0.31 ng-N g−1 per week in respective control treatments. Simultaneous heterotrophic nitrification and aerobic denitrification (SHNAD) was found to be a prominent bioprocess at low temperature that resulted in high and stable total nitrogen and nitrate accumulation from garden soil and goat manure with relative recovery efficiency of 11.12%, 14.97% and 14.20%; 19.34%. A. caligenosa shows promising prospects for mass applicability in biogenic N removal from manure of Himalayan goat. [ABSTRACT FROM AUTHOR]

Beauveria bassiana is one of the most widely studied and used entomopathogenic fungus as biopesticide. In the biological control of pests, B. bassiana will persist in the soil after application, and will inevitably contact with earthworms, especially the epigeic earthworm species. So, what are the effects of earthworm and its epidermal mucus on the activity of B. bassiana? We employed the epigeic earthworm Eisenia fetida, B. bassiana TST05 strain, and the insect Atrijuglans hetaohei mature larvae to study the impact of earthworm epidermal mucus on the vitality and pathogenicity of B. bassiana to insect. Methods included scanning electron microscope observation, detection of spore germination, fungal extracellular enzyme activity, and infection testing to A. hetaohei. The results showed that the B. bassiana spores may attach to the cuticle of E. fetida but they could be covered by the epidermal mucus and became rough and shrunken. After treatment with the epidermal mucus, the spore germination and extracellular enzymes of B. bassiana was significantly inhibited. Inoculation of A. hetaohei larvae with a mixture of B. bassiana and mucus showed that the mucus could reduce the pathogenicity of B. bassiana to the insect, resulting in a slower disease course and lower mortality. It was concluded that the epidermal mucus of the earthworm E. fetida can inhibit the activity of B. bassiana, as well as the infectivity and pathogenicity of fungus to target insects. However, after treatment with epidermal mucus the surviving B. bassiana still had certain infectivity to insects. This is of great significance for the application of B. bassiana in biological control of pests. [ABSTRACT FROM AUTHOR]

In recent years, animals and plants have received increasing attention as potential next-generation protein production systems, especially for biopharmaceuticals and animal proteins. The aim of the present study was to develop the earthworms Eisenia fetida Waki and Eisenia andrei Sagami as next-generation animal protein production hosts. These earthworms have been approved as model animals for acute toxicity tests by the Organization for Economic Co-operation and Development, and they have post-translational modification systems. However, so far, none of the studies have used earthworm transfection techniques. Thus, we developed a transfection method for E. fetida and E. andrei using microinjection and electroporation systems. The maximum survival rates and transfection efficiencies were 79.2% and 29.2% for E. fetida, and 95.8% and 50.0% for E. andrei, respectively. Furthermore, human erythropoietin was detected in the transformed earthworm tail fragments using an enzyme-linked immunosorbent assay. These results contribute to the development of a potential earthworm-based novel animal protein production system. [ABSTRACT FROM AUTHOR]

Rice (Oryza sativa L.) straw, an agricultural waste of high yield, is a sustainable source of fermentable sugars for biofuel and other chemicals. However, it shows recalcitrance to microbial catalysed depolymerization. We herein describe development of thermotolerant microbial consortium (RSV) from vermicompost with ability to degrade rice straw and analysis of its metagenome for bacterial diversity, and lignocellulolytic carbohydrate active enzymes (CAZymes) and their phylogenetic affiliations. RSV secretome exhibited cellulases and hemicellulases with higher activity at 60 °C. It catalysed depolymerization of chemical pretreated rice straw as revealed by scanning electron microscopy and saccharification yield of 460 mg g−1 rice straw. Microbial diversity of RSV was distinct from other compost habitats, with predominance of members of phyla Firmicutes, Proteobacteria and Bacteroidetes; and Pseudoclostridium, Thermoanaerobacterium, Chelatococcus and Algoriphagus being most abundant genera. RSV harboured 1389 CAZyme encoding ORFs of glycoside hydrolase, carbohydrate esterase, glycosyl transferase, carbohydrate binding module and auxiliary activity functions. Microorganisms of Firmicutes showed central role in lignocellulose deconstruction with importance in hemicellulose degradation; whereas representatives of Proteobacteria and Bacteroidetes contributed to cellulose and lignin degradation, respectively. RSV consortium could be a resource for mining thermotolerant cellulolytic bacteria or enzymes and studying their synergism in deconstruction of chemically pretreated rice straw. [ABSTRACT FROM AUTHOR]

Northeastern Thailand relies on agriculture as a major economic activity, and has used high levels of agrochemicals due to low facility, and salty sandy soil. To support soil recovery and sustainable agriculture, local farmers have used organic fertilizers from farmed animal feces. However, knowledge about these animal fecal manures remains minimal restricting their optimal use. Specifically, while bacteria are important for soil and plant growth, an abundance and a diversity of bacterial composition in these animal fecal manures have not been reported to allow selection and adjustment for a more effective organic fertilizer. This study thereby utilized metagenomics combined with 16S rRNA gene quantitative PCR (qPCR) and sequencing to analyze quantitative microbiota profiles in association with nutrients (N, P, K), organic matters, and the other physiochemical properties, of the commonly used earthworm manure and other manures from livestock animals (including breed and feeding diet variations) in the region. Unlike the other manures, the earthworm manure demonstrated more favorable nutrient profiles and physiochemical properties for forming fertile soil. Despite low total microbial biomass, the microbiota were enriched with maximal OTUs and Chao richness, and no plant pathogenic bacteria were found based on the VFDB database. The microbial metabolic potentials supported functions to promote crop growth, such as C, N and P cyclings, xenobiotic degradation, and synthesis of bioactive compounds. Pearson's correlation analyses indicated that the quantitative microbiota of the earthworm manure were clustered in the same direction as N, and conductivity, salinity, and water content were essential to control the microbiota of animal manures. [ABSTRACT FROM AUTHOR]

Abstract The role of earthworms in soil carbon dynamics is a recent avenue of research which is less studied in India. Three plots of 1 m3 size were laid in Jeevaka live laboratory (JLL)- a biodiversity rich area within the University campus. A control plot (CP) of same dimension was maintained outside JLL. Out of three plots within JLL, one was operated with native earthworm Perionyx ceylanensisMichaelson (100 numbers), water and cattle dung as feed (Jeevaka test plot- JT) and fenced with nylon mesh. Remaining two plots were operated as controls within JLL (JC1 and JC2). JC1 (Jeevaka control 1) was provided with cattle dung and water, while JC2 and CP (outside JLL) were operated without any supplements. Throughout the experiment native earthworm species have maintained their dominancy in all plots except CP where no earthworms were observed. At the end of a year-long study, JC1 with maximum diversity of earthworms showed better soil organic carbon (SOC) and particulate organic carbon (POC)-which is relatively a stable form of SOC. Overall findings indicate better the diversity of earthworms better is the carbon storage in the soil.

In the present study rice straw (R, control) was mixed with Cowdung (C), Azolla (A) and cellulolytic fungus Aspergillus terreus (F) in different combinations viz. RC, RA, RF, RCF, RCA, RFA and RCFA and subjected to aerobic composting (Acom) and vermicomposting (Vcom - with Eisenia fetida). It was found that addition of azolla and cattledung to two parts straw(RCA-666: 314:20 g) caused fastest degradation (105 days), gave maximum population buildup of E. fetida (cocoons, hatchlings and worm biomass), highest decline in pH, EC, TOC and C/N ratio and maximum increase over control in N(17.72%), P(44.64%), K(43.17%), H (7.93%), S (14.85%), Ca(10.16%), Na(145.97%), Fe(68.56%), Zn(12.10%) and Cu(32.24%). Rice straw (R) took longest time for degradation i.e. 120 and 140 days and had lowest content of nutrients in Vcom as well as Acom group. RCFA was also converted into Vcom at the same time but other parameters were less than RCA except for highest content of B (19.87%), Mg(21.27%) and Mn (5.58%). Bioconversion of three parts straw (RCA-735:245:20 g) was also faster (110 days) with vermicomposting than all the mixtures of Acom group (130-140 days) but nutrient content was slightly less than RCA with 2 parts straw. The results show that azolla reduces dependence on cattledung for recycling the carbon rich rice straw and enhances its agronomic value. [ABSTRACT FROM AUTHOR]