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Present study deals with cocoon biology of tropical epigeic earthworm Perionyx excavatus (Perrier), Perionyx ceylanensis (Michaelsen) and Eudrilus eugeniae (Kinberg). Ultra structure observed under Scanning Electron Microscope (SEM) revealed that wall of the cocoon is porous (which helps in aeration) and the terminal part of the cocoon is softer than that of middle part which facilitates the hatching process. The size of the cocoon varies among the species and is related to the diameter and length of the clitellum. All the epigeic species under study are continuous breeder with high fecundity. Among the studied species shortest incubation period, highest fecundity and highest hatching rate were observed in P. ceylanensis. Cocoon production in these three epigeic species increases in summer and monsoon season and decreases in the winter season. Temperature is the limiting factor for cocoon production as well as incubation period of cocoon. Cocoon production was affected by temperature and body weight of the earthworm. Thus it can be considered that cocoon production in epigeic earthworm species depends on biological factor (body mass), as well as, ecological factor (temperature). Continuous breeding strategy, high fecundity, high rate of cocoon production, as well as high hatching rate, indicates greater usefulness of P. ceylanensis followed by E. eugeniae than P. excavatus in the vermiculture based biotechnology in North-East India.

Aim: The aim of the present study was to observe the role of cerebral ganglionic neurosecretory cells (NSCs) during cocoon production in native earthworm species Perionyx ceylanensis subjected to amputation and seasonal variations. Methodology: Histological studies (using Aldehyde Fuchsin and Chrome Alum Haematoxylin Phloxin stain) were carried out on brain NSCs in the two groups of earthworms (Group I and Group II) maintained in earthen culture pots (2L) with cowdung as food. Group I comprised of brain amputed earthworms was subjected to observe the role of brain NSCs in production of cocoon and Group II earthworms subjected to seasonal changes in the cerebral NSCs during cocoon production. Five replications were kept for Group I (1 individual per pot) and Group II (1 pair per pot). Results: Group I debrained earthworms started to lay cocoons from the 31st day following regeneration of cerebral ganglionic type A NSCs. In group II worms the highest neurosecretory activity was registered in the cerebral type A cells, especially during monsoon coinciding with the hike of cocoon generation. Interpretation: Appearance of type A NSCs in regenerated brain and peak of type A neurosecretory cell activity during peak reproductive period of earthworm species (as indicated by peak of cocoon production) indicates the possible role of cerebral type A NSCs in cocoon laying.

The data on growth and reproduction of composting earthworms may be beneficial for large scale earthworm production. The growth and reproduction of Perionyx excavatus were assessed in limited supplies of four experimental diets-cow dung alone and its mixture with acacia (Acacia auriculiformis) leaf litter, bamboo (Bambusa polymorpha) leaf litter and terrestrial weed Mikania micrantha under laboratory conditions to select suitable diet from our locally available resource for vermiculture. Growth performance of Perionyx excavatus was significantly (P

An experiment was carried out to determine the effect of replacing dietary fish meal protein with earthworm meal protein on growth performance, whole-body composition, and hematology of butter catfish (Ompok pabda). This experiment was conducted in 15 glass aquaria (200 L) with five treatments, each having three replications. Each of the aquaria was stocked with 40 fishes (average weight 0.62 g). The fish were fed with iso-nitrogenous diets having 35% protein (21% protein from fish meal and/or earthworm meal) twice a day up to satiation with the experimental diets, where fish meal protein was replaced by earthworm meal protein at 0% (control), 25%, 50%, 75%, and 100% in the treatments E0, E25, E50, E75, and E100, respectively. At the end of the 8-week rearing period, the best growth performances including weight gain (5.23 ± 0.14 g), percent weight gain (844.15 ± 6.38%), and specific growth rate (4.00 ± 0.11% day−1), and best feed utilization parameters, viz., feed conversion ratio (1.57 ± 0.03) and protein efficiency ratio (2.87±0.04), were found in treatment E75, where 75% of fish meal protein in the diet was replaced with earthworm meal protein. The hematological parameters showed no abnormality due to supplementation of earthworm meal in the diets. At the end of the experiment, whole-body carcass composition, viz., protein, lipid, and ash contents (15.93 ± 0.19%, 7.22 ± 0.60% and 2.14 ± 0.10%, respectively), of O. pabda was higher in treatment E75 compared to control (14.8 2± 0.34, 6.16 ± 0.27, 1.89 ± 0.10, respectively). The results imply that earthworm meal can be used as a protein-rich source to replace fish meal, the major protein source, in the diet of O. pabda up to 75% level without compromising growth, immunity, and body composition.

The ecological suitability of different leaf litter types in enhancing the biological diversity and micronutrient status in soil needs to be understood for deciding the plant species to be used for afforestration practices. The present research work was done to study the colonization of an epigeic earthworm, Perionyx excavatus in the decomposing leaf litter of five locally important tree species- Cashew, Shal, Mango, Acacia and Eucalyptus up to a period of 90 days. The major nutrient and anti-nutrient chemical parameters of the leaf litters were compared and rates of degradation of the selected nutrient and anti-nutrient parameters were studied in laboratory microcosms. P. excavatus showed maximum colonization for Cashew followed by Shal where both these tree species exhibited comparatively lower anti-nutrient content and significantly higher rates of degradation of the selected nutrient parameters. Lowest colonization by P. excavatus was observed in case of Eucalyptus which exhibited highest polyphenol and tannin content and significant lower rates of degradation of all the selected nutrient parameters and hence their utilization in microcosms with P. excavatus. The results indicated that lower content of tannins and polyphenols in case of Cashew and Shal can be related to the palatability of these leaf litters and thereby exhibiting maximum colonization by P. excavatus. Thus, Cashew and Shal can be considered as suitable tree species for afforestation practices for enhancing nutrient recycling and effective soil conservation.

Graphical abstract, Highlights • Coelomic fluid of 3 earthworms exhibited cytotoxicity on oral cancer cells. • Coelomic fluid induced DNA strand breakage in SCC-9 cells. • Coelomic fluid of Perionyx excavatus showed highest percentage of G2M cycle arrest. • Coelomic fluid of Eudrilus eugeniae caused a 37-fold rise in apoptosis percentage., The current protocol of cancer management includes surgery, radiotherapy and chemotherapy. However, these modalities have significant adverse effects and affect the quality of life. Further intensification of treatment is hindered as maximal toxicity levels are reached impeding improvement. Hence researchers are in the quest for adjunctive naturally available therapies that can alter tumor proliferation without causing significant adverse reactions. The present study aims to explore the cytotoxic potential of earthworm coelomic fluid (ECF) of Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) on oral cancer cell line SCC-9. The effect of ECF on cell cycle analysis and mechanism of cell death have also been investigated. All experiments reported in this paper were performed as 3 replicates per experiment. The results indicated that ECF of EE, EF and PE have potent variable cytotoxic effect on SCC-9 cells demonstrated through LDH, clonogenic and comet assay. An effective cell cycle arrest was observed at the G2M phase of cell cycle with apoptotic induction that was observed through an Annexin V – FITC/PI assay. ECF of EE was found to be superior in its cytotoxic action closely followed by ECF of PE. The present findings provide evidence for the first time that ECF of EE, EF and PE have potent cytotoxic effect on oral cancer cells in vitro. They significantly induce G2M cell cycle arrest and promote apoptosis in SCC-9 cell line. Gene expression studies have been planned to ascertain the pathways of cell death.

Fifteen days pre-decomposed seaweeds, Halimeda gracilis, Gracilaria corticata, Sargassum wightii and Sargassum swartzii spiked with cowdung (1:1) were vermicomposted using Perionyx excavatus for 60 days. The pH in the vermicompost showed insignificant reduction while electrical conductivity showed significant enhancement (P 0.05). The reduction of organic carbon in vermicomposts ranged from -37.78 to -50.97% over worm-unworked composts. Total NPK contents showed significant increment (26.72-78.17%) in vermicompost over worm-unworked composts. The difference in percentage increase/decrease between physicochemical parameters was statistically significant (P 0.001) and the same pattern was found between substrates. The total microbial population in vermicomposts was significantly higher than that of initial and composts of all seaweed + cowdung combinations (P 0.001). The growth and reproduction of Perionyx excavatus in seaweed + cowdung combinations showed equivalent or higher rates when compared with cowdung signifying that Perionyx excavatus is well suited to convert seaweed and cowdung combinations into nutrient rich vermicompost.

The seaweeds, Gracilaria edulis, Turbinaria ornata and Ulva reticulata collected from Mandapam East Coast region of Tamil Nadu, India has been subjected to vermicomposting in combination with cowdung (CD) in 1:1 combination for 60 days under laboratory conditions. The percentage decomposition was ranged between 83.70 and 87.00 in the vermicompost of seaweed + CD combinations while it was 30.71 and 35.12% in worm-unworked compost where the decomposition rate of substrates with earthworms is significantly (P

The present study demonstrates the vermistabilization of paper mill wastewater sludge (PMS) spiked with cow dung (CD) employing indigenous epigeic earthworm Perionyx excavatus Perrier. A total of six treatments were prepared along with a positive control (PC) and negative control (NC). Twenty earthworms were released into each treatment including PC and NC without earthworms. The different proportions viz., T1 (CD – 100%), T2 (PMS:CD – 1:3), T3 (PMS:CD – 1:2), T4 (PMS:CD – 1:1), T5 (PMS:CD – 3:1), T6 (PMS:CD – 2:1), PC (PMS – 100%) and NC (PMS – 100%) and changes in chemical parameters and microbial properties were recorded during the course of 60 days. Vermistabilization caused a significant decrease in the level of heavy metals: Cd (2.9–27.8%), Cu (0.22–42.3%), Pb (1.3–56.3%) and Cr (0.8–46.2%). The bioconcentration factor (BCFs) was also calculated and great amount of heavy metals accumulated in their body (mg kg−1) that ranged from 0.31 ± 0.003–0.45 ± 0.007 for Cd, 0.12 ± 0.005–0.24 ± 0.003 for Cu, 0.15 ± 0.005–0.31 ± 0.006 for Pb and 0.29 ± 0.007–0.56 ± 0.001 mg kg−1 for Cr, accumulation of heavy metals are in the order: Cr > Cd > Pb > Cu. The physicochemical parameters of earthworm treated substrate such as electrical conductivity, total nitrogen, total phosphorus and total potassium were significantly increased; whereas, pH, total organic carbon, C:N ratio (carbon: nitrogen) and C:P ratio (carbon: phosphorus) were reduced after 60 days of vermistabilization. The vermistabilized materials also had a higher population of bacteria (98.90 ± 0.30 CFU × 106 g−1), fungi (43.75 ± 0.55 CFU × 103 g−1) and actinomycetes (67.65 ± 0.45 CFU × 105 g−1) than initial mixtures. Moreover, several histopathological changes were observed in earthworm tissues viz., disintegration of cells, irregular surface of epidermis, cellular debris, irregular cellular compartmentation, and oval-shaped nucleus. Higher level of histopathological abnormalities was recorded in PC (PMS-100%) while none were detected in lower concentrations of PMS. Furthermore, the study concludes that the paper mill sludge in a mixture of cow dung (1:1 ratio) can be a useful proposition for utilizing this hazardous waste through the adoption of vermitechnology.

Carbon to nitrogen (C/N) ratio influences substrate combinations and earthworm performance in vermicomposting systems. To elucidate these factor effects, a comparative evaluation of species, C/N ratio combined with feed rate, was conducted on three local earthworm species: Perionyx excavatus, Eudrilus eugeniae, and Dichogaster annae. Earthworms were stocked at similar densities and fed shredded paper (SP), cattle manure (CM), and lawn clippings (LC) combined to form C/N ratios of 28, 36, and 53. Earthworms were fed at rates of 1, 1.25, and 2 g feed (dry wt.)/g worm/day for a period of 8 weeks. Percent vermiconversion, earthworm adult and juvenile biomass, and vermicast quality were measured. Vermicast production was significantly affected by the combination of C/N ratio and feed rate and varied among species. All treatment combinations resulted in > 70% conversion, except E. eugenaie fed at the medium rate. Vermiconversion increased for P. excavatus and D. annae with increasing C/N ratio but decreased with increasing the feed rate. Vermicast EC, pH, and C/N ratio was strongly affected by species, relative to other experimental factors. D. annae showed the greatest change in biomass, which peaked at the highest feed rate and lowest C/N ratio. Average adult biomass decreased for P. excavatus with increasing feed rate, while differences were nonsignificant for E. eugenaie and D. annae. Significant increases in average juvenile biomass were only evident for D. annae in response to increasing feed rates. Feed rate had a greater influence on earthworm population dynamics and vermicast quality compared to initial feedstock C/N ratio.

The earthworm Perionyx excavatus is a species highly capable of bidirectional regeneration. Pair-rule genes are thought to have an ancestral function in arthropod segmentation. However, orthologs in annelids (i.e. Capitella teleta) do not exhibit segmental expression in the ectoderm or mesoderm. Their role in regeneration is currently unclear. Here, we report the expression profile of primary pair-rule genes (Pex-EvxA, Pex-EvxB, Pex-RuntA, Pex-RuntB, Pex-Hes1A, Pex-Hes1B, Pex-Hes4A, and Pex-Hey) found in P. excavatus using semi-quantitative reverse transcription-polymerase chain reaction. Our results indicated these genes showed variable expression during bidirectional regeneration. Six of these genes might play diverse and potentially critical roles in head and/or tail regeneration.

Tri-n-butyl phosphate (TBP) is widely used in various industrial processes and has been detected in all environmental matrices. So far, little work has been done regarding the metabolism of TBP on terrestrial invertebrates. We investigated the metabolism of TBP in the earthworm, Perionyx excavatus, after acute exposure to TBP for one and two days in filter paper contact test, as well as after chronic exposure for 28 days in soil experiment. Biotransformation products were identified by using liquid chromatography coupled with quadrupole time-of-flight mass spectrometry, and by exploiting the information dependent acquisition in tandem mass spectrometry. TBP exhibited low accumulation in earthworm-soil ecosystem at 10 mg/kg and 50 mg/kg. The presence of earthworms significantly enhanced TBP degradation at 50 mg/kg in soil. Dibutyl phosphate and hydroxylated TBP were the major phase I metabolites. Three novel phase II metabolites were identified: ethanol dibutyl phosphate and its sulfate conjugate, and the phosphate conjugate of hydroxylated TBP. Hydroxylation and further phosphorylation dominated metabolism in chronic exposure. An extensive metabolic pathway of TBP in earthworm was proposed. This is the first report of TBP metabolism in terrestrial invertebrates and highlights the necessity to identify metabolites of contaminants when evaluating their bioaccumulation and toxicity.

In this study, an analysis of organic fertilizer of an agro-waste (Sesame straw) plus cow dung was carried out using an epigeic earthworm species Eisenia fetida and Perionyx sansibaricus. Sesame straw is abundantly produced after each harvesting of the crop in Kharif season in arid region of tropical India. The compost produced in presence and absence of earthworm exhibit significant (P0.05) changes in physicochemical properties respectively. In control bedding, the values of water holding capacity enhanced significantly (P

The present study is incorporated with agrowaste management and biofertilizer production in tropical India. For this, an exotic earthworm species Eisenia fetida and a local species Perionyx sansibaricus were used to decompose cumin straw and cow dung and evaluated vermicomposting efficacy. Physicochemical parameters of the vermibed such as pH, electrical conductivity, water holding capacity, organic carbon, C/N ratio, total nitrogen, phosphorus and potassium were evaluated in the vermicompost. The values of all the above physicochemical parameters of E. fetida worked bedding material varied significantly (PPerionyx sansibaricus also varied significantly (PE. fetida was better than P. sansibaricus for cumin plus cow dung bedding. Though E. fetida is better agrowaste converter but both the earthworm species may be used for organic waste management in desert region. It may provide the support in the form of biofertilizer to enhance the organic farming in region of the Thar Desert.

Biological and behavioral responses of the tropical earthworm Perionyx excavatus towards different concentrations of abamectin were evaluated. Abamectin significantly reduced the biomass and reproduction (cocoon production) of P. excavatus as well as inducing histopathological alterations in the cuticle. Biomass loss was recorded in P. excavatus exposed to abamectin at a concentration as low as 0.1 mg active ingredient (a.i.) kg−1, while atrophy, another physiological response, was observed at an abamectin concentration of 0.21 μg cm−2 in a filter paper test. Cocoon production was significantly reduced in the presence of abamectin, and no cocoons were produced at doses of 20 mg a.i. kg−1 or higher, while abamectin at 50 mg a.i. kg−1 induced extreme pathology, characterized by the loss of the integrity of the whole body wall and intestine of P. excavatus. Histopathological alterations can be used as a biomarker to evaluate the toxicological impact of exposure to abamectin.

Fetal Bovine Serum (FBS) is used as a major supplement in culturing animal cells under in vitro conditions. Due to ethical concern, high cost, biosafety, and geographical as well as batchwise result variations, it is important to reduce or replace the use of FBS in animal cell culture. The major objective of this work is to evaluate the feasibility of heat-inactivated coelomic fluid (HI-CF) of the earthworm, Perionyx excavatus as a possible alternative for FBS in animal cell culture experiments. The coelomic fluid (CF) was extruded from the earthworm using electric shock method and used for the experiments. Electric shock method is a simple non-invasive technique, which has no harmful effect on earthworms. Mouse primary fibroblast and HeLa cell lines were used in this study. Among HI-CF, autoclaved CF and crude CF, the supplement of medium with HI-CF shows positive results. The processed HI-CF (90°C for 5 min) at 10% supplement in cell culture medium promote maximum cell growth but cells need the initial support of FBS for the attachment to the culture flask. Microscopic observation and immunofluorescence assay with actin and lamin A confirm that the cellular and molecular morphology of the cells is maintained intact. The HI-CF of earthworm, P. excavatus has shown better cellular viability when compared with FBS and making it possible as an alternative supplement to minimize the use of FBS.

Disease outbreaks are being increasingly recognized as a significant constraint on aquaculture production and trade affecting the economic development of the sector in many countries. Extracting and using biologically active compounds from earthworms has traditionally been practiced by indigenous people throughout the world. The aim of the present study was to shown antimicrobial activity through earthworm extract against fish bacterial pathogens. In total, 8 bacterial strains i.e. 6 gram negative viz. Aeromonas hydrophila, Pseudomonas aeruginosa, P. fluorescens, E.coli, Enterobacter aerogens and Shigella sp. and 2 gram positive viz. Staphylococcus aureus and Micrococcus luteus were identified. The extract of earthworm Perionyx excavatus, Pheretima posthuma were prepared and antimicrobial activity of the extract was determined by antimicrobial well diffusion assay. After 24 hrs of incubation period, it was observed that earthworm extract showed antibacterial activity against isolated bacterial strains. Among earthworm extract of two different species, the maximum zone of inhibition was shown against A. hydrophila by Perionyx excavatus (18.33± 0.66 mm) and P. posthuma (16.66±0.33). P. excavatus showed antibacterial activity against all pathogenic bacteria except Shigella spp. However on the other hand, P.posthuma showed antibacterial activity against A. hydrophila, P. fluorescens, E.coli, and S. aureus. The study has proved that earthworm extract can be effectively used for suppression of bacterial infection in fishes and that it can used as potential antimicrobial drug against commercial antibiotic resistance bacteria.

The aim of this study was to recycle and reuse the enormously available unutilized lignocellulosic solid organic waste resource, cashew leaf litter (CLL) admixed with various animal dungs, cowdung, sheepdung and horsedung by employing predominantly available indigenous epigeic earthworm—Perionyx excavatus (Perrier, 1872) and produce quality vermifertilizer. Four different combinations of each [(100 % dung alone, 3:1 (75 % dung + 25 % CLL), 2:2 (50 % dung + 50 % CLL) and 1:3 (25 % dung + 75 % CLL)] vermibeds were allowed for vermicomposting process under laboratory conditions. After 60 days, the worm worked vermicompost and worm unworked normal compost were harvested and characterized. The earthworm activity—growth, reproductive performance (cocoon production and hatchling number) and recovery of vermicompost was also studied. The obtained results clearly showed that vermicompost from CLL admixed with cowdung at 2:2 ratio had lower pH, organic carbon, C–N ratio, C–P ratio, lignin, cellulose, hemicellulose and phenol content, and higher nitrogen, phosphorus, potassium dehydrogenase and humic acid content than the raw substrates and worm unworked normal compost. In addition, pronounced and better earthworm activity was found in the above combination. Through vermitechnology way of producing agronomic valid vermicompost using natural waste resources like CLL and animal dungs can be used as bio-organic fertilizer. These vermiresources have vast and diversified potential for maintaining sustainable soil health, fertility, productivity, waste degradation, soil reclamation, land restoration practices and environment health.

Globally, 1.3 billion tonnes of food produced for human consumption are wasted each year and are usually discarded in landfills where the greenhouse gas, methane, is produced during decomposition. Vermicomposting is a possible organic waste management strategy, which may reduce the amount of food waste disposed in landfills and produce compost, which is a usable end product. This study aimed to evaluate how efficiently the Perionyx excavatus can convert organic waste into quality vermicompost.

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.

Water hyacinth is well known for accumulating heavy metals from the water bodies and its uncontrolled growth; due to which it is posing serious threat to the water bodies and the environment. Therefore, the present paper aimed at the safe treatment and recycling of nutrients by vermicomposting of water hyacinth. Three different earthworm species E. fetida, E. eugeniae, P. excavatus were compared for its higher biomass growth, organic matter degradation and heavy metal changes during the study. The nutrient concentration and stability parameters (OUR and CO2 evolution) were evaluated during the degradation of water hyacinth in five different trials. Results showed that nutrients (Na, K, Ca, Mg, total Nitrogen (TN) and available Phosphorous (AP)) were increased and the stability parameters were reduced significantly at the end of vermicomposting. It was observed that E. eugeniae accumulated higher concentration of heavy metals in its body tissue during transit of waste through worm’s gut. On an overall, E. fetida proved to be the best on account of biomass increment as well as juveniles hatched when compared with the other earthworm species during the treatment.

Epigeic earthworms can significantly hasten the decomposition of organic matter, which is known to be mediated by gut associated microflora. However, there is scanty information on the abundance and diversity of the gut bacterial flora in different earthworm genera fed with a similar diet, particularly Eisenia foetida and Perionyx excavatus. In this context, 16S rDNA based clonal survey of gut metagenomic DNA was assessed after growth of these two earthworms on lignocellulosic biomass. A set of 67 clonal sequences belonging to E. foetida and 75 to P. excavatus were taxonomically annotated using MG-RAST and RDP pipeline servers. Highest number of sequences were annotated to Proteobacteria (38-44%), followed by unclassified bacteria (14-18%) and Firmicutes (9.3-11%). Comparative analyses revealed significantly higher abundance of Actinobacteria and Firmicutes in the gut of P. excavatus. The functional annotation for the 16S rDNA clonal libraries of both the metagenomes revealed a high abundance of xylan degraders (12.1-24.1%). However, chitin degraders (16.7%), ammonia oxidizers (24.1%) and nitrogen fixers (7.4%) were relatively higher in E. foetida, while in P. excavatus; sulphate reducers and sulphate oxidizers (12.1-29.6%) were more abundant. Lignin degradation was detected in 3.7% clones of E. foetida, while cellulose degraders represented 1.7%. The gut microbiomes showed relative abundance of dehalogenators (17.2-22.2%) and aromatic hydrocarbon degraders (1.7-5.6%), illustrating their role in bioremediation. This study highlights the significance of differences in the inherent microbiome of these two earthworms in shaping the metagenome for effective degradation of different types of biomass under tropical conditions.

Helicases are known to be a proteins that use the chemical energy of NTP binding and hydrolyze to separate the complementary strands of double-stranded nucleic acids to single-stranded nucleic acids. They participate in various cellular metabolism in many organisms. DEAD-box proteins are ATP-dependent RNA helicase that participate in all biochemical steps involving RNA. DEAD-box3 (DDX3) gene is belonging to the DEAD-box family and plays an important role in germ cell development in many organisms including not only vertebrate, but also invertebrate during asexual and sexual reproduction and participates in stem cell differentiation during regeneration. In this study, in order to identify and characterize DDX3 gene in the earthworm, Perionyx excavatus having a powerful regeneration capacity, total RNA was isolated from adult head containing clitellum. Full length of DDX3 gene from P. excavatus, Pe-DDX3, was identified by RT-PCR using the total RNA from head as a template. Pe-DDX3 encoded a putative protein of 607 amino acids and it also has the nine conserved motifs of DEAD-box family, which is characteristic of DEAD-box protein family. It was confirmed that Pe-DDX3 has the nine conserved motifs by the comparison of entire amino acids sequence of Pe-DDX3 with other species of different taxa. Phylogenetic analysis revealed that Pe-DDX3 belongs to a DDX3 (PL10) subgroup of DEAD-box protein family. And it displayed a high homology with PL10a, b from P. dumerilii.

The aim of the study was to examine safe reuse and recycling of organic waste digestate obtained from a biogas plant (5 % total solid) with enhanced nutrient value through vermitechnology. Two indigenous epigeic earthworm species Perionyx excavates and Perionyx sansibaricus were tried individually for this purpose. The results clearly show a significant increase in pH values from 6.5–7.4, electrical conductivity (21.3–21.7 %), total N (84.5–94.6 %), total P (35.9–47.1 %), total K (49.8–52.6 %), Ca (41.9–41.9 %) and a significant decrease in total organic C (17.1–22.4 %), C/N ratio (7.2–6.9), C/P ratio (20.3–20.6), COD (51.9–55.7 %), BOD (85.5–91.2 %). Similarly, indicator organisms (fecal coliforms and fecal streptococci) showed decrement at the end of the composting period (60 days). Fecal Coliforms reduce to nil, while in fecal streptococci a 6 log reduction was observed. Oxygen uptake rate dropped to 67.4–70 % for vermireactors. Overall, the aforementioned findings highlighted that the indigenous earthworm species could enhance the nutrient value of the anaerobic digestate, which could be utilized as an efficient soil conditioner.

India has a large network of pulp and paper mills of varying capacity. On an industrial scale the sludge from paper and pulp mills is disposed of either as landfill or incinerated. Both methods result in the loss of a valuable resource and have obvious environmental and economic disadvantages. The solid waste from pulp and paper mills is a source of organic matter and its proper disposal and management is the responsibility of the industry. As composting/vermicomposting could be used to transform this waste trials were carried out to determine the feasibility of converting dewatered sludge (DS) into a value added end product using an earthworm, Perionyx excavatus. The vermicomposting of the waste resulted in an increase in its electrical conductivity (EC), ash content, total nitrogen (TN), total phosphorous (TP) and available phosphorous (AP), respectively, and a decrease in total organic carbon (TOC), biochemical oxygen demand (BOD), chemical oxygen demand (COD), oxygen uptake rate (OUR) and evolution of carbon dioxide (CO2). Overall, the best treatment was T5 in which there was a 76.1% increase in TP, 58.7% in TN, 74.5% decrease in TOC , and a reduction of 6.7 fold in the production of CO2 and 10.7 fold in BOD, respectively. Our trials demonstrate that vermicomposting using an epigeic earthworm, Perionyx excavatus, is an alternate and environmentally safe way of recycling paper mill sludge if it is mixed with an appropriate amount of cow dung and food processing waste. Overall T5 was the best combination of paper mill sludge and waste for vermicomposting followed by T3, T2, T4 and T1, respectively.

Epigeic earthworms (Oligochaeta) have been appeared as key organisms to convert organic waste resources into value-added products, i.e., vermicompost and worm biomass. The assessment of reproduction potential of composting earthworm may be beneficial for large-scale earthworm production. Although, the waste minimizing potential of Perionyx excavatus and Perionyx sansibaricus is well proved, but little information is available about their fecundity rate. In this study, the efforts have been made to explore the growth and reproduction biology of P. excavatus and P. sansibaricus, using cattle waste solid as culture substrate, under laboratory conditions. Earthworms were weighed weekly and number of cocoons produced per week assessed. Biomass productions, fecundity, maturation, natality all were significantly different between P. excavatus and P. sansibaricus. The highest mean individual biomass was 767.7 ± 18.4 mg and 612.6 ± 20.6 mg, respectively in P. sansibaricus and P. excavatus. However, the highest cocoon numbers occurred in P. excavatus (492.3 ± 13.6), significantly higher than P. sansibaricus (269.6 ± 17.1). Fecundity was slightly different in both species: 1.38 ± 0.77 cocoons adult worm−1 week−1 (P. excavatus) and 1.58 ± 0.74 cocoons adult worm−1 week−1 (P. sansibaricus). The hatchling success rate (%) was highest in P. excavatus. Overall natality (juveniles adult−1 week−1) was highest in P. sansibaricus (1.52) than P. excavatus (1.26), which suggests that P. sansibaricus may be a better candidate for rapid propagation of earthworms in cattle waste solid.

The impact of organic material quality on biomass production and reproduction potential of commercial composting earthworm species: Eudrilus eugeniae, Perionyx excavatus and Perionyx sansibaricus were studied, by using three different type of culture material namely Jbs: (Jowar straw (Sorghum vulgare ) + bajra straw ( Pennisetum typhoides ) + sheep manure) (1:1:2), fym (farmyard manure), and Kw + Ll (Kitchen waste + leaf litter of Magifera indica ) (1:1), under laboratory conditions for 150 days. The above substrate or culture materials have different palatability, particle size and physio- chemical composition. Kitchen waste (C-to-N = 26.7) as well as farmyard manure (C-to-N = 27.4) is a high quality material with fast decomposition rates, while crop residues are low quality materials with slow decomposition rates (C-to-N = 45.6). All the three studied earthworm species showed maximum biomass production rate in Kw + Ll culture ( E. eugeniae = 9.80 ± 0.01 mg worm -1 day -1 , P. excavatus = 3.75 ± 0.01 mg worm -1 day -1 , and P. sansibaricus = 3.77 mg worm -1 day -1 ). Individual cocoon production rate varied drastically, and maximum value (worm -1 week -1 ) of it was observed in Kw + Ll for E. eugeniae (1.88 ± 0.15) and P. sansibaricus (1.77 ± 0.14), while P. excavatus showed maximum cocoon production rate in Fym (1.79 ± 0.17). The hatchling success of cocoons obtained from different beddings was also observed and cocoon obtained from Kw + Ll culture exhibited maximum hatchling success (%) in all the three species studied. The cocoons of both E. eugeniae and P. sansibaricus , obtained from Fym culture showed the highest number of hatchlings (cocoon -1 ) i.e. 1.59 ± 0.04 and 1.81 ± 0.10, respectively, whereas cocoons of P. excavatus showed the highest hatchling number (1.77 ± 0.06) in Kw + Ll. In this present study, there was a consistent trend of decreasing individual biomass as well as cocoon production rate, followed by their peak values with ageing of the culture materials. The relationship between different earthworm parameters and N-content or C-to-N of culture material was also evaluated. The biomass production rate and hatching success in all the three species studied showed direct correlation (p

The research of applying Ganoderma residue in earthworm Perionyx excavatus cultivation have promised in reducing environmental pollutions from mushroom farm and brought high economic efficiency. A 9 m square with 9 experimental formulas were applied to investigate the effect of Ganoderma residue to earthworm growth. The results showed that Perionyx excavatus increased mass in all formulas added cultivated Ganoderma residues, in which the largest increased in formula of 75 % cultivated Ganoderma residues treated with Trichoderma, from 0.59 kg/m2 initial to 2.62 kg/m2. In addition, Perionyx excavatus growth and development completely was seen on the medium with 100 % cultivated ganoderma residues, reached 1.89 kg/m2. Although the growth rate of the Perionyx excavatus cultivated with Ganoderma residues was slower as compared to farming entirely by cow manure (2.94 kg/m2), in the long run, this process will bring farmers higher profits (56.500 VN dongs /m2 of culturing). This cultivation process might be promising to be applied in regions with mushrooms farms for economic and environmental reasons.

In this study, (1)H nuclear magnetic resonance (NMR)-based metabolomics approach was used to characterize the metabolic response of the earthworm Perionyx excavatus in continuous vermifiltration for two months under hydraulic loading rates of 1m(3)m(-2)d(-1) (VF1) and 1.5m(3)m(-2)d(-1) (VF1.5). Both VF1 and VF1.5 showed higher removal of chemical oxygen demand and total nitrogen than the biofilter without earthworms. Principal component analysis of the NMR spectra of earthworm metabolites showed significant separations between those not subjected to wastewater filtration (control) and VF1 or VF1.5. Temporal variations of earthworm biomass, and the identified metabolites that are significantly different between control, VF1 and VF1.5 revealed that worms underwent increasing metabolic activity within 20days in VF1 and 14days in VF1.5, then decreasing metabolic activity. The use of NMR-based metabolomics in monitoring earthworm metabolism was demonstrated to be a novel approach in studying engineered vermifiltration systems.

Vermicomposting is proven to be an effective way for nutrient cycling, converting large quantities of organic waste into value added product (vermicast). In order to harness the potential of earthworms in vermicomposting, selection of earthworm species that are able to consume large quantity of waste, and moreover, produce vermicast with high nutrient content is important. This experiment was carried out to compare the efficacy of Perionyx excavatus (PE) and Eudrilus eugeniae (EE) in vermicomposting rice straws. Ten earthworms were introduced into each vermibin containing grinded rice straw. The vermicast produced was collected periodically. The experiment was terminated when 70% of rice straw had decomposed. The plant nutrient contents and humic acids in vermicast were analysed. Vermicast of PE contained higher concentrations of total and available N, P, K and Mg while EE vermicast contained higher total and available Ca. Humic acid content was also found to be higher in EE vermicast. EE took 134 days while PE took 171 days to complete vermicomposting, thus, plant nutrient content generated per day in vermicast EE was higher compared to PE. Using EE in vermicomposting would contribute significantly towards practicing sustainable agricultural by recycling large amount of organic waste rice straw into value added high plant nutrient content vermicast. Key words: Earthworm biomass, decomposition rate, humic acid, agronomy.

Annelida is a lophotrochozoan phylum whose members have a high degree of diversity in body plan morphology, reproductive strategies and ecological niches among others. Of the two traditional classes pertaining to the phylum Annelida (Polychaete and Clitellata), the structure and function of the Hox genes has not been clearly defined within the Oligochaeta class. Using a PCR-based survey, we were able to identify five new Hox genes from the earthworm Perionyx excavatus: a Hox3 gene (Pex-Hox3b), two Dfd genes (Pex-Lox6 and Pex-Lox18), and two posterior genes (Pex-post1 and -post2a). Our result suggests that the eleven earthworm Hox genes contain at least four paralog groups (PG) that have duplicated. We found the clitellates-diagnostic signature residues and annelid signature motif. Also, we show by semi-quantitative RT-PCR that duplicated Hox gene orthologs are differentially expressed in six different anterior–posterior body regions. These results provide essential data for comparative evolution of the Hox cluster within the Annelida.

Terrestrial avoidance behavior is proposed as a fast and cost-effective method for assessing effects of pesticides on earthworms. Tropical species however, have rarely been used in avoidance tests. Avoidance tests were performed with Perionyx excavatus, a tropical species, and Eisenia andrei as the standard species, using chlorpyrifos and carbofuran in artificial and natural soil. Earthworms were exposed to concentrations of 1–900 (chlorpyrifos) and 1–32 (carbofuran) mg a.i. kg−1 dry soil in a two-chamber system under tropical conditions (26 ± 2 °C, 48 h). No significant difference was found in the control tests comparing the two soils used, suggesting soil type did not affect the distribution of the worms. The results suggest a higher sensitivity of E. andrei, with EC50S for the effect on avoidance behavior for both pesticides being a factor of 2–3 lowers than for P. excavatus. Earthworm avoidance tests with local species should therefore be used with caution when applied as a tool for pesticide risk assessment in the tropics. Endpoints generated through avoidance tests in this study are shown to be less sensitive than reproduction and more sensitive than survival. This was further confirmed by literature data available. Earthworm avoidance tests therefore can only replace survival tests as an initial screening tool for risk assessment.

A serine collagenolytic protease was purified from the earthworm Perionyx excavates by DEAE-Sephadex A-50 and DEAE-Sephadex A-50 column. The molecular mass of the earthworm serine protease was estimated to be 35 kDa. The purified serine protease was optimally active at pH 6.8-7.8 and 55°C. In vitro studies on collagenolytic activity of this enzyme was also carried out using type I collagen. Hydrolysis of collagen using serine protease was characterized by the collagenolytic activity. The amount of hydroxyproline released (μg/ml) on untanned and tanned leather was monitored. Zymographic analysis was carried out using Collagen as substrate.

The lignocellulosic wastes, cashew leaf litter (CLL) admixed with various animal dungscowdung (CD), sheepdung (SD) and horsedung (HD) have been subjected to vermicomposting by using indigenous earthworms, Perionyx excavatus and Lampito mauritii and to produce value added vermicompost. Four different combinations of each [100% dung alone, 3 : 1 (75% dung + 25% CLL), 2 : 2 (50% dung + 50% CLL) and 1 : 3 (25% dung + 75% CLL)] vermibeds were prepared for vermicomposting process. After 60 days, the vermicompost and worm-unworked normal compost were harvested and characterized and growth (biomass), reproductive performance of earthworms-cocoon production, hatchling number and recovery of vermicompost were also studied. The obtained results clearly showed that vermicompost from CLL admixed with CD at 2 : 2 ratio had lower pH, organic carbon, C–N ratio, C–P ratio, lignin, cellulose, hemicellulose and phenol content, and higher nitrogen, phosphorus, potassium, dehydrogenase and humic acid content than the raw substrates and normal compost. Also pronounced and better earthworm activity was found in the above combination for both species. Hence organic farming practices using vermicompost promise an alternative to the existing practices and proved as a sustainable approach.

The composting potential of two epigeic earthworms (P. excavatus and P. sansibaricus) was studied in 2002 to breakdown the domestic waste under laboratory conditions. The experimental container with P. sansibaricus showed maximum mineralization and decomposition rate than that of P. excavatus. Except for exchangeable K (it was higher (P = 0.004) in a container with P. excavatus), the domestic waste processed by P. sansibaricus showed about 6% more total nitrogen (P = 0.002) and about 7% more available P (P = 0.269) at the end than by P. excavatus. As compared with the initial level organic C content as well as C:N ratio showed a considerable reduction that was noted higher in substrate with P. sansibaricus than those by P. excavatus (organic C, t-test: P = 0.870; C:N ratio, t-test: P = 0.002). The growth (biomass increase) and reproduction parameters such as mean individual live weight, maximum individual growth rate (mg wt./worm/day), number of cocoons and reproduction rate (cocoon/worm/day) were higher in bedding with P. sansibaricus. The maximum earthworm mortality was in vermibed having P. sansibaricus (∼50% higher than by P. excavatus) (t-test: P = 0.423), since both species did not show a drastic difference in waste mineralization rate, but comparatively, P. excavatus exhibited better growth and reproduction performance, which further support the suitability of the species for large scale vermiculture operations.

An earthworm assay was used to assess the toxicity of two main gasoline components. Ecotoxicities of toluene and methyl tert -butyl ether (MTBE) were compared for Asian earthworm Perionyx excavatus separately and in combination. Sum of toxic unit (TU) at 50% mortality for the mixture (LC50 mix ) was estimated from the dose (TU-based)–response relationships by the Trimmed Spearman–Karber method. Toluene was shown to be about five and two times more toxic to P. excavatus than MTBE in filter paper contact test and natural soil tests, respectively. This is primarily due to the adsorption of toluene to soil particles, leading to less bioavailability of toluene to the earthworm in soil. The combined effect of toluene and MTBE was investigated using toxic unit model, and it was found to be almost additive and synergistic responses to P. excavatus in filter paper contact test and soil tests, respectively. The combined effect of toluene and MTBE in filter paper contact test was not consistent with the results in soil toxicity tests. This phenomenon may be associated with the interaction of soil salts with pollutants. Treatment with toluene and MTBE also affected the behavior and morphology of P. excavatus . It indicates that the VOCs induce metabolic and functional damages in earthworms. Combined effects of gasoline components should be taken into account to soil risk assessment.

In view of the growing awareness about vermicomposting technology in recycling different types of organic wastes, this study was conducted to investigate the effect of initial substrate pH on vermicomposting. The substrate pH and ash content were evaluated as a function of time. The data showed an exponential relationship between substrate pH and time of vermicomposting while a phase Bode plot of a single zero system relationship between the ash content and time of vermicomposting. The model parameters of these relationships also had very good correlation with the initial substrate pH. On the basis of obtained correlation between model parameters and initial substrate pH, generalized predictive models for the substrate pH and ash content have been evolved in terms of the duration of vermicomposting and the initial substrate pH. Plots of the predictive and experimentally observed values indicated a high robustness of predictive models. The study also revealed that the earthworm species Perionyx excavatus performs well in a wide range of substrate pH. Near neutral initial substrate pH was found to be optimal for stabilization of waste with minimal processing time. The substrates having strong acidic initial pH were found to be less suitable for vermicomposting.

A laboratory scale study was conducted to assess the suitability of rubber leaf litters as vermiculture substrates for Perionyx excavatus, Eudrilus eugeniae and Eisenia fetida. Earthworm mortality, their rate of body weight increase and reproduction were measured during the investigation. In E. eugeniae, E. fetida and P excavatus, the rate of increase of body weight was 28.8, 6.2 and 5.04 mg worm - 1 day - 1 , respectively, in rubber leaf litter. Mortality of the adults was very high (50 %) in P. excavatus when compared with that (17 %) of both Eisenia and Eudrilus. Rate of reproduction in rubber leaf litter diet for both E. eugeniae (1.4 young worm - 1 week - 1 ) and E. fetida (1.3 young worm - 1 week - 1 ) was significantly higher (P E. fetida > P. excavatus.

Summary A laboratory scale study was conducted to assess the suitability of rubber leaf litters as vermiculture substrates for Perionyx excavatus, Eudrilus eugeniae and Eisenia fetida. Earthworm mortality, their rate of body weight increase and reproduction were measured during the investigation. In E. eugeniae, E. fetida and P. excavatus, the rate of increase of body weight was 28.8, 6.2 and 5.04 mg worm–1 day–1, respectively, in rubber leaf litter. Mortality of the adults was very high (50%) in P. excavatus when compared with that (17%) of both Eisenia and Eudrilus. Rate of reproduction in rubber leaf litter diet for both E. eugeniae (1.4 young worm–1 week–1) and E. fetida (1.3 young worm–1 week–1) was significantly higher (P E. fetida > P. excavatus.

Biomass production and reproduction of a group of four adult Perionyx excaratus were studied in limited supplies of four experimental diets; cowdung alone and its mixtures with straw, bamboo leaf litter or kitchen waste, in order to select a suitable diet medium for vermiculture. P. excavatus showed maximum rate of biomass increase and reproduction in the mixtures with straw and bamboo leaf litter. In spite of achieving the highest final biomass value. P. excavatus showed the lowest rate of biomass increase and reproduction in the mixture with kitchen waste. Cowdung, a natural food of P. excavatus, was marginally better than the mixture with kitchen waste with regard to the rate of biomass increase and reproduction.

A study was conducted to evaluate the ash content as a function of time of vermicomposting and the moisture content which plays a significant role in the process. The data revealed a linear relationship between the ash content and time in the effective range of the vermicomposting duration. The model parameters of this linear relationship also have linear correlation with the moisture content. Through sub- stitutions, a generalized predictive model for the ash content has been evolved in terms of the duration of vermicomposting and the moistures contents. A plot of the predictive and experimentally observed values indicated a high robustness of predictive model. The study also showed that a moisture content of 80% is optimum for stabilization of waste in minimum processing time.

A comparative study was performed to evaluate the breakdown of leaf litter of two broad-leaved tree species, i.e. Polyalthia longifolia and Rhododendron arboreum by an exotic earthworm species Eisenia foetida and an indigenous earthworm species Perionyx excavatus under laboratory condition. Methods of Haimi and Huhta (Biol Fertil Soil 10:178–183, 1990) were followed for the present study. The rate of decay, percent cellulose, hemicelluloses and lignin contents of the decomposing leaf litter of P. longifolia and R. arboreum were found to be higher in the sets treated with the two different earthworm species as compared to that of untreated sets where no earthworm was added. Though there was not much difference in the rate of leaf litter breakdown by two different earthworm species, the rate of litter breakdown by the exotic earthworm, E. foetida, was slightly higher as compared to the indigenous earthworm P. excavatus. Both the selected leaf litters treated with the different earthworm species exhibited significant positive correlations between percent biomass remaining, cellulose, hemicelluloses and lignin contents (p ≤ .001). Significant variation was observed in hemicelluloses of leaf litters in sets treated with the two different earthworm species and in the untreated sets. It can be concluded that the indigenous earthworm species P. excavatus also has a high potential for leaf litter breakdown. It can act as a potential candidate in vermicompost technology—which will be of immense help in wastes management, resource recovery and environmental conservation.

Perionyx excavatus is a potential earthworm species for vermicomposting, and of live feed for fish as it contains high quality protein with excellent amino acid profile. Its limited seasonal abundance in natural as well as the associated risk of bioaccumulation of toxic products in wild earthworm inflate large scale vermiculture of this species. To establish a suitable food composite for mass culture of that species, the present study evaluated the production performance and nutritional composition of P. excavatus fed on four different organic food composites namely, C, T 1, T 2 and T 3. Initially five adult wild earthworms were stocked in cemented tanks, and reared them with feeding respective organic composites at the rate of 10% of their body weight twice a week for 120 days. To determine the growth rate, newly hatched larvae of 0.12-0.16 mg collected from respective treatments was reared in the separate tubs at the stocking rate of 5 larvae tub -1 for 60 days. Temperature, pH, moisture, total nitrogen and organic carbon contents of rearing beds, and growth rate of earthworms were recorded at every fortnight. The treatment T 3 was found to be effective for production performance (2.508 ± 0.75 larvae week -1 ) while the T 2 gave higher growth rate (0.16 g·worm -1 ·week -1 ) to this earthworm species. The highest protein (48.36%) and moisture content (81.85%) were recorded in T 2 but T 3 contained the lowest protein (39.65%) and the highest lipid level (4.82%). The ash content was found not to be significantly different among all treatments. The results of this study demonstrated T 3 composition as good medium for reproduction, and T 2 composition as better food composite for rearing P. excavatus . The outcomes of this study may provide essential information towards the mass culture of this species.