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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 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

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.

In this study, we isolated 28 strains of Bacillus spp. from water samples, catfish pond mud samples and earthworm manure (Perionyx excavates). By the cross-streak agar methods, 22 Bacillus strains showed the inhibition ability to Edwardsiella ictaluri, which caused Bacillary Necrosis Pangasius (BNP) in catfish (Pangasianodon hypophthalmus). Both Bacillus sp. Q16 and Q111 strains showed the highest inhibition to E. ictaluri by the double-layer agar methods. Finally, two Bacillus strains (Q16, Q111) were selected as a source of potential probiotic because of the ability of extracellular enzyme secretion (protease, amylase, cellulose) strong growth at 0,1-1% salt concentrations, survival within the pH range 6-8, resistance to low pH and low bile salts, inability to produce haemolysin enzyme, sensitivity to eight antibiotics in the three impacting groups (inhibition of wall synthesis, inhibition mechanism of protein synthesis, inhibition of nucleic acid synthesis). Two Bacillus strains (Q16, Q111) were identified that they belong to Bacillus subtilis by biochemical method and 16S rRNA gene sequencing method. This study indicated that two Bacillus strains (Q16, Q111) isolated from catfish pond can be applied as high potential probiotics that used to farm catfish.

Objective: The purpose of the experiment was to examine the effect of different food sources on the growth and reproduction performances of an epigeic earthworm Perionyx excavatus. Materials and Methods: The experiment was carried out in 18 cylindrical plastic containers for 10 weeks. The study was designed with six treatments, each having three replicates. In control treat¬ment (T1), cow dung was used as the only food source for the earthworm. In another five treat¬ments, water hyacinth (T2), chopped banana plant trunk (T3), vegetable scrap (T4), paddy straw (T5), and sugarcane bagasse (T6) were used as food sources with cow dung as bedding material. Results: The maximum weight gain of earthworm P. excavatus was 3,294.7 ± 4.5 mg for the food staff of vegetable scrap (T4). Earthworm P. excavatus fed with chopped banana plant trunk (T3) showed a very similar weight gain of 3,243.7 ± 3.8 mg. On the contrary, the minimum weight gain was 1,799.7 ± 3.5 mg for the food staff of paddy straw (T5). The maximum cocoon number of 137.33 ± 6.46 mg was observed in T3, whereas a minimum number of 36.67 ± 4.16 mg in T1. The highest number of hatchlings (12.33 ± 0.88 mg) was recorded in treatment T3, whereas the least number of hatchling (5.00 ± 0.58 mg) was observed in T1. Conclusion: It was concluded that, among the six different food sources, chopped banana plant trunk was preferable food source to the earthworm for growth and reproduction. [J Adv Vet Anim Res 2020; 7(2.000): 331-337]

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.

PURPOSE: Main insight of the study was to evaluate the potential of Perionyx excavatus, an indigenous earthworm of India towards bioconversion of toxic paper mill wastes. The study had considered a comprehensive study over succession of different enzymes and microbial biomasses alongside removal of heavy metals to reflect the qualitative enrichment of the toxic waste through valorisation procedure. METHODS: The process of vermicomposting was performed using ratio of paper mill sludge: cow dung: straw as 5:4:1 and Perionyx excavatus for a period of 60 days. Qualitative enrichment was considered through detailed evaluation of physicochemical parameters and succession of 19 enzymes using API ZYM technology. Heavy metal loads in terms of Zn, Cr, Cu, Pb along with bioaccumulation factor (BAF) were considered to establish functional role of earthworm in bioremediation. Microbial succession was measured in terms of PLFA biomarkers. RESULTS: Enzymatic enrichment of different isozymes of esterase, aminopeptidase and glycosyl hydrolase were found in the vermicompost. A significant reduction of heavy metals was noticed in vermicomposting of primary sludge as Zn > Pb > Cr > Cu and secondary sludge as Zn > Pb > Cu > Cr. An increase in density of Gram positive bacterial and fungal populations was found at the end phase of vermicomposting process which supports maturity of vermicompost. CONCLUSION: Presence of different enzyme activities, reduction of heavy metal loads along with rise of eukaryotic PLFA markers, had illustrated the efficiency of vermicomposting as a valorisation procedure in the qualitative enrichment of toxic organic paper mill sludge.

Incorporation of vermin culture in the composting system produces “vermicompost”, an enriched biofertilizer known to improve the physical, chemical, and biological properties of soil. It is applied in granular form and/or in liquid solution (vermiwash), and in both open fields and greenhouses. Vermicompost has been shown to contain plant growth hormones, which stimulate seed germination and improve crop yield, the ‘marketability’ of products, plant physiology, and their ability to fight against disease. In recent years, South Pacific island countries (SPICs) have placed an increasing emphasis on the importance of organic agricultural practices as a means of achieving more sustainable and environmentally friendly farming practices. However, vermiculture is not practiced in South Pacific island countries (SPICs) largely due to the lack of awareness of this type of application. We consider the inclusion of vermiculture in this region as a potential means of achieving sustainable organic agricultural practices. This study represents a systematic review in which we collect relevant information on vermicomposting and analyze the applicability of this practice in the SPICs based on these nations’ physical, socioeconomic, and climatic conditions. The tropical climate of the SPICs means that they meet the combined requirements of a large available biomass for composting and the availability of earthworms. Perionyx excavatus and Pontoscolex corethrurus have been identified as potential native earthworm species for vermicomposting under the conditions of the SPICs. Eisenia fetida, a well-known earthworm species, is also effectively adapted to this region and reported to be an efficient species for commercial vermicomposting. However, as a new input into the local production system, there may be unforeseen barriers in the initial stages, as with other advanced technologies, and the introduction of vermiculture as a practice requires a steady effort and adaptive research to achieve success. Further experimental research is required to analyze the productivity and profitability of using the identified native earthworm species for vermiculture using locally available biomass in the SPICs.

Paddy is one of the most important food crops in India. In Tripura of north east India, paddy covers 78–90% of cultivable areas with hydromorphic clay-rich acidic soils. As earthworms have important role in above-ground crop production, the present study deals with the earthworm communities in the soils of wet land paddy agro-ecosystem of West Tripura (India). The study was conducted during May 2015 to August 2017. Earthworms were collected from the paddy fields by TBSF monolith (25 × 25 × 30 cm) digging and hand sorting method. Temperature, moisture, pH and organic matters were estimated from 0 to 15 cm depth soils. Biological parameters such as density, biomass, relative abundance and ecological parameters such as indices of diversity, dominance etc. were measured. A total of 7 earthworm species belonging to 4 families and 5 genera were recorded: [Almidae Glyphidrilus sp. (near to gangeticus)], Moniligastridae (Drawida assamensis, Drawida papillifer papillifer and Drawida sp.), Megascolecidae (Metaphire houlleti, Perionyx excavatus) and Glossoscolecidae (Pontoscolex corethrurus). The average density and biomass of earthworms in the paddy plantations were 163 ind. m−2 and 56 g m−2 respectively. Diversity and dominance indices of earthworms of paddy plantations were 0.46 and 0.60 respectively. Glyphidrilus sp. contributed more than 50% biomass and density of earthworm community and probably had a great role in paddy production due to enormous deposition of casts (‘cast hills’) in the rhizosphere zone of paddy plants.

Recovering the nutrients from organic solid residues is an ever demanding approach in terms of eco-friendly management. In this study, the efficiency of the earthworms, Perionyx ceylanensis and Perionyx excavatus in recycling and recovering plant nutrients from urban and agricultural biomass residues was investigated. Vermicomposting of four different types of organic biomass resources in combination with cowdung (1:1) for 50 days was conducted using the earthworms, P. ceylanensis, and P. excavatus. Physico-chemical, microbiological and maturity characteristics of the final product were analyzed. Results showed a decrease in pH, organic matter content (OMC), total organic carbon (TOC), cellulose, lignin, carbon to nitrogen (C/N) and carbon to phosphorus (C/P) ratios, and an increase in total Kjeldahl nitrogen (TKN), phosphorus (TP), potassium (TP), calcium and sodium in vermicomposts over composts (prepared without earthworms). TKN increase in vermicomposts over composts was insignificant between P. ceylanensis and P. excavatus, while it was significant for TP, TK, and total microbial population (P

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

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.

VERMIWASH is a dark-colored liquid fertilizer, which is collected after water passes via an earthworm culture column. It is rich in plant growth hormones, micro-and maco-nutrients like N, P, and K. An experiment was conducted using three earthworms’ types; Allopophora spp, Eisenia fetida and Perionyx excavates to produce vermiwash. Some chemical and microbial analyzes were performed (pH, EC, macro-and microelements, toxic metals, total count of aerobic bacterial, fungi, actinomycetes, total coliform, salmonella and shigella). The results indicated that pH of vermiwash was slightly alkaline and recorded 7.14 followed by 7.35 and 7.46 with the Allopophora spp, Eisenia fetida, and Perionyx excavatus, respectively. The electrical conductivity (EC) recorded an insignificant difference between Perionyx excavatus and Eisenia fetida it was 3.7 and 3.6 dS m-1. Using Perionyx excavatus and Eisenia fetida provided the highest total N values; 9044 and 9040 mg L-1 respectively, while the total P and K were the highest content (2627 and 2607mg L-1, respectively) when Perionyx excavatus was used. The concentration of Fe and Cu were the highest when using Perionyx excavatus with values of 16.4 and 0.21 mg L-1 respectively. The Mn recorded its highest concentration in M, Allopophora sppas 0.50 mg L-1. The vermiwash content of Cd, Ni, and Co was not detected, while the low Cr concentrations were recorded with the vermiwash resulting from the three earthworms. As well, the vermiwash obtained from the Perionyx excavatus was the highest in the total aerobic bacterial count, while Allopophora spp provided the best values for each total number of fungi and actinomycetes with an absence of total coliform, Salmonella and Shigella in all tested samples.

Earthworms are ecological engineers that can contribute to the displacement of biological control agents such as the entomopathogenic nematodes (EPNs) and fungi (EPF). However, a previous study showed that the presence of cutaneous excreta (CEx) and feeding behavior of the earthworm species Eisenia fetida (Haplotaxida: Lumbricidae) compromise the biocontrol efficacy of certain EPN species by reducing, for example, their reproductive capability. Whether this phenomenon is a general pattern for the interaction of earthworms-entomopathogens is still unknown. We hypothesized that diverse earthworm species might differentially affect EPN and EPF infectivity and reproductive capability. Here we investigated the interaction of different earthworm species (Eisenia fetida, Lumbricus terrestris, and Perionyx excavatus) (Haplotaxida) and EPN species (Steinernema feltiae, S. riojaense, and Heterorhabditis bacteriophora) (Rhabditida) or EPF species (Beauveria bassiana and Metarhizium anisopliae) (Hypocreales), in two independent experiments. First, we evaluated the application of each entomopathogen combined with earthworms or their CEx in autoclaved soil. Hereafter, we studied the impact of the earthworms' CEx on entomopathogens applied at two different concentrations in autoclaved sand. Overall, we found that the effect of earthworms on entomopathogens was species-specific. For example, E. fetida reduced the virulence of S. feltiae, resulted in neutral effects for S. riojaense, and increased H. bacteriophora virulence. However, the earthworm P. excavates increased the virulence of S. feltiae, reduced the activity of H. bacteriophora, at least at specific timings, while S. riojaense remained unaffected. Finally, none of the EPN species were affected by the presence of L. terrestris. Also, the exposure to earthworm CEx resulted in a positive, negative or neutral effect on the virulence and reproduction capability depending on the earthworm-EPN species interaction. Concerning EPF, the impact of earthworms was also differential among species. Thus, E. fetida was detrimental to M. anisopliae and B. bassiana after eight days post-exposure, whereas Lumbricus terrestris resulted only detrimental to B. bassiana. In addition, most of the CEx treatments of both earthworm species decreased B. bassiana virulence and growth. However, the EPF M. anisopliae was unaffected when exposed to L. terrestris CEx, while the exposure to E. fetida CEx produced contrasting results. We conclude that earthworms and their CEx can have positive, deleterious, or neutral impacts on entomopathogens that often coinhabit soils, and that we must consider the species specificity of these interactions for mutual uses in biological control programs. Additional studies are needed to verify these interactions under natural conditions., MC is supported by a Moroccan scholarship of the Ministry of National Education, Vocational Training, Higher Education and Scientific Research, and the travel assistance associated with the grant CSIC ICOOP + 2018 grant (COOPA20231). RBP is financed with an FPI contract (CAR-2018) by the Department of Economic Development and Innovation of The Government of La Rioja (Spain). IVD is supported with an FPI-UR fellowship by the Universidad de La Rioja (Spain). RCH is awarded by Ramon y Cajal contract award (RYC-2016-19939) of the Government of Spain. This study is supported by CSIC I-COOP + 2018 grant (COOPA20231).

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.

Regeneration is a biological process restoring lost or amputated body parts. The capability of regeneration varies among organisms and the regeneration of the central nervous system (CNS) is limited to specific animals, including the earthworm Perionyx excavatus. Thus, it is crucial to establish P. excavatus as a model system to investigate mechanisms of CNS regeneration. Here, we set up a culture system to sustain the life cycle of P. excavatus and characterize the development of P. excavatus, from embryo to juvenile, based on its morphology, myogenesis and neurogenesis. During development, embryos have EdU-positive proliferating cells throughout the whole body, whereas juveniles maintain proliferating cells exclusively in the head and tail regions, not in the trunk region. Interestingly, juveniles amputated at the trunk, which lacks proliferating cells, are able to regenerate the entire head. In this process, a group of cells, which are fully differentiated, reactivates cell proliferation. Our data suggest that P. excavatus is a model system to study CNS regeneration, which is dependent on the dedifferentiation of cells.

This communication deals with the morphology, taxonomy and life history of six new acephaline gregarines (Apicomplexa: Gregarinomorphea) of the genus Monocystis Stein (1848), Nematocystis Hesse (1909), Aikinetocystis Gates (1926) and Dirhynchocystis Cognetti de Martiis (1921) from oligochaete hosts. These are Monocystis molnari sp. nov. from Metaphire anomala (Clitellata: Opisthopora: Megascolecidae); Monocystis paneki sp. nov. from Lampito mauritii (Clitellata: Opisthopora: Megascolecidae); Monocystis bulbae sp. nov from Metaphire posthuma (Clitellata: Opisthopora: Megascolecidae); Nematocystis cylindrum sp. nov. from Perionyx excavatus (Clitellata: Opisthopora: Megascolecidae); Aikinetocystis perionyxae sp. nov. from Perionyx excavatus (Clitellata: Opisthopora: Megascolecidae) and Dirhynchocystis drawidae sp. nov. from Drawida nepalensis (Clitellata: Haplotaxida: Moniligastridae). Trophozoites of Monocystis molnari sp. nov. elongated and cylindrical. Gametocysts elliptoid. Oocysts fusiform. Trophozoites of Monocystis paneki sp. nov. elongated and hump like. Gametocysts orbicular. Oocysts fusiform. Trophozoites of Monocystis bulbae sp. nov. elongated and cylindrical. Gametocysts fusiform. Oocysts fusiform. Trophozoites of Nematocystis cylindrum sp. nov. cylindrical. Gametocysts orbicular. Oocysts fusiform. Trophozoites of Aikinetocystis perionyxae sp. nov. cylindrical. Gametocyst ovoid. Oocysts fusiform. Dirhynchocystis drawidae sp. nov. having a capsule like body. Gametocysts orbicular. Oocysts rhomboid.

Background Earthworm communities are generally very sensitive to physico-chemical properties of the soil in different agro-ecosystem i.e. cultivated or non-cultivated which directly or indirectly influence the earthworm survival. The difference in physico-chemical properties of soil at different sites contributed to the formation of population patches for earthworm species. Understanding the physico-chemical properties of soil at a particular site could facilitate the prediction of earthworm species at that site. The objective of the present study was to investigate the diversity, abundance, and distribution of earthworms in cultivated and non-cultivated agroecosystems and their physico-chemical properties affecting the earthworm diversity and abundance. Results Total 10 species of earthworms i.e. Amynthas alexandri, Amynthas morrisi, Eutyphoeus incommodus, Eutyphoeus waltoni, Metaphire birmanica, Metaphire houlleti, Metaphire posthuma, Octochaetona beatrix, Perionyx excavatus, and Polypheretima elongata, were reported. Out of all the reported species, Metaphire posthuma was found to be the most abundant earthworm species in both cultivated and non-cultivated agroecosystems with the occurrence at 56.81% sites. The Shannon-Wiener index (H), Margalef species richness index (DMg) and Pielou species evenness (E) was ranged from 0 to 0.86, 0 to 0.64 and 0.78 to 1 respectively. The principal component analysis resulted in four principal components i.e. PC1, PC2, PC3 and PC4 which contributing variance (%) of 22.96, 19.37, 14.23 and 10.10 respectively. The principal component analysis also showed that physico-chemical parameters of soil such as EC, pH, TDS, texture, OC, moisture, etc. play a critical role in earthworm distribution. Conclusion The conventional farming system has a negative effect on the earthworm diversity in the soil while the physico-chemical properties of soil also have a determinant effect on the same. Earthworms abundance in the present study have significant direct relation with soil properties at a particular site and vice versa. The diversity indices also change due to the conventional farming system which directly affects the earthworm abundance.

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.

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.

Vermicomposting is usually performed with one earthworm species. However, use of a consortium of two or more species (Eisenia fetida, Eudrilus eugeniae, and Perionyx excavatus) is rare. Research on optimization of vermicomposting duration is also insufficient. Three earthworm species in various combinations were fed with cowdung mixed biowaste feedstock. Organic C, C/N ratio, compost respiration, coliform count, toxic metals, and alkalinity reduced at maturity; whereas, earthworm biomass, NPK availability, enzyme activity, microbial growth, and humic substances remarkably improved under consortium systems. Thus, the harvest quality was optimized sooner under vermicomposting (40–60 days) than composting (>100 days). Phospho-lipid fatty-acid (PLFA) analyses revealed the enriched variations in microbial community structure and fatty-acid profiles in consortium treated vermicomposts. Overall, the Eisenia+Eudrilus+Perionyx consortium produced the best quality compost in the shortest duration (40–50 days) followed by Eisenia+Eudrilus and Eisenia+Perionyx consortia. Tomato seed germination and root-shoot vigors were significantly greater in consortia mediated systems. Moreover, economic assessment confirmed the advantage of consortium mediated 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 current study was carried out for the purpose of comparative evaluation of vegetable market waste degradation potential of three earthworm species (Eisenia fetida, Perionyx excavatus and Eudrilus eugeniae). The vegetable waste used as substrate in this study was collected from Mission Chariali vegetable market in Tezpur, Assam, India. Non-biodegradable materials such as polythenes, plastic etc. were segregated and organic waste was sun-dried. Cow dung was mixed with the substrate in 1:3 proportion and subjected to vermicomposting with three earthworms species separately. The data on physicochemical properties of the compost bed along with earthworm activity were collected every 10 days interval. The experiment was continued until 70 days. Analysis of the data indicated that E. fetida posed maximal nutrient mineralization and reduction in C:N ratio when compared with the data of day 0 with of day 70. Also, the values of vermicomposting coefficient were maximum for E. fetida. Moreover, the mean individual weight of E. fetida was also maximum during the decomposition period. Although Eu. eugeniae increased in total biomass much more rapidly than E. fetida, nevertheless, the cocoon production rate was greatest (78.67) in case of E. fetida, indicating the differential resource allocation with trade-off between sex alloca-tion and growth by Eu. eugeniae in a non-conducive environment during its life cycle. Overall, the results indicate suitability of E. fetida for vegetable waste degradation.

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.

Agricultural utilisation is one of the most promising uses of sewage sludge in Hungary. Sewage sludge can be applied to agricultural fields in two ways: the injection of dewatered sewage sludge and the application of sewage sludge after composting. Vermicomposting is a special type of composting, where the organic residues are broken down by earthworms. The worms facilitate the decomposition process both by mixing the sludge and by physically degrading it. Earthworm species have various morphotypes requiring different habitats. Compost worms have great adaptability to extreme conditions and are capable of exploiting organic matter in a state of decomposition. Eisenia sp., Eudrilus eugeniae and Perionyx excavatus are important species for vermicomposting. When examining the role and possibilities of vermicomposting, it is important to compare it with traditional composting methods. The most important aspect of producing vermicompost is to ensure optimum environmental conditions for the earthworms, especially in terms of temperature, humidity and aeration, which requires constant attention. An important feature of traditional composting is the thermophilic phase, during which the pathogenic organisms in sewage sludge are destroyed. The thermophilic phase is omitted during vermicomposting due to the thermal sensitivity of the earthworms, but the presence and activity of the earthworms results in similar sterility. Regarding its nutrient content, vermicompost contains larger quantities of total and plant-available macroelements than conventional composts. A further advantage is the presence of the plant hormone agents excreted by earthworms. From the environmental point of view, the ability of earthworms to accumulate heavy metals and the role of their special gut flora in the decomposition of organic pollutants could contribute to the wider use of vermicomposting to dispose of sewage sludge. While vermicompost has many advantages, a number of obstacles need to be overcome before it can be routinely used in Hungary. Many landowners regard sewage sludge compost as hazardous waste that could contaminate their soil and crops rather than as a nutrient and soil amendment. Although numerous studies have been published on sewage sludge, the assessment of long-term effects, including the issues currently of most concern in Hungary, is still lacking. Vermicomposting is therefore a promising, innovative technology for sewage sludge recycling. Sewage sludge and sewage sludge composts with pollutant contents greater than the limits laid down in Government Regulation 50/2001. (IV.3.) can be made suitable for agricultural use by vermicomposting.

Environmental improvement is an accepted national goal. Under the present condition of the acute energy crisis and environmental degradation with rise in population, it has become essential to develop an appropriate technology for the recovery of energy from non-conventional sources like organic wastes. India produces 350 million tonnes of organic wastes from different agricultural sources. Out of these, several million tonnes of plant nutrients are produced. Earthworm (verm) is known to be a good biological source for the recovery of vermifertilizer and vermiprotein from the organic wastes to be used in agro-ecosystem and aquaculture and poultry, respectively. Vermicomposting is a process of conversion of organic waste through the synergistic actions of epigeic earthworm and bacteria. When earthworms (endogeic or anecic) are applied for breakdown and stabilization of municipal solid wastes, the process is termed ‘vermistabilization’. The ultimate goal of both the processes is the production of plant-available nutrients and earthworm biomass (vermiprotein). Earthworm species to be selected for the process (1) should have great adaptability with respect to environmental factors, (2) should be capable of inhabiting high percentage of organic matter, (3) should be a prolific breeder (high fecundity) with high hatching success, and (4) should have short life cycle. Recent studies indicated that under Indian conditions epigeic species, such as Perionyx excavatus, Perionyx ceylanensis, Eudrilus eugeniae, Eisenia fetida, Eisenia andrei, can complete their life cycle in organic waste and produce vermicompost. Lampito mauritii, Polypheretima elongata, Drawida nepalensis, Drawida willsi, Metaphire posthuma, Metaphire houlleti can degrade organic matter in municipal sewage sludge which contains considerable amounts of sand particles. This chapter deals with the growth and reproduction of these organic waste degrading earthworms. Based on the study, suitable species for organic waste degradation are being suggested.

Spent mushroom substrate (SMS) is a recalcitrant lignocellulosic waste. Recycling of SMS through composting has been reported; however, the process is lengthy due to its complex biochemical composition. Although vermitechnology is known for its high efficiency, it has rarely been applied to recycle SMS. In this study, the qualitative value of vermicomposted SMS mediated by three earthworm species (i.e., Eisenia fetida, Eudrilus eugeniae, and Perionyx excavatus) was evaluated on the basis of nutrient availability, microbial activity, phospholipid fatty acid (PLFA) profiles, and seed germination assays. Degradation profiles of the lignocellulosic substrate in the vermireactors were assessed by monitoring the changes in crystallinity and distribution of functional groups using X-ray diffraction (XRD) and Fourier transform infrared spectroscopy, respectively. Total organic carbon decreased by 1.4–3.5 folds with approximately 2.1–2.4 folds increase in nitrogen and phosphorus availability in all vermibeds. Interestingly, pH declined in the Eisenia and Eudrilus systems but increased in the Perionyx-vermibeds. XRD-derived crystallinity index was reduced significantly by 1.37 folds in Perionyx-vermicompost with concurrent microbial enrichment. Further, profuse abundance of vital functional groups (C O, NH, and OH) was clearly observed in the vermicompost with Perionyx followed by that with Eisenia. Moreover, PLFA illustrated significant variations in fatty acid distributions and microbial communities of the three vermicomposting systems. The seed germination assay showed that the germination index and relative root-shoot vigor of Perionyx-vermicompost treated seeds were 1.05–1.30 times greater than those of Eisenia and Eudrilus vermicompost treated ones. The results suggest that SMS degradability was affected by the growth of a healthy microbial community through vermicomposting.

The process of bioconversion of solid organic wastes through vermicomposting justifies the environmental message for sustainability such as reduce, recycle and reuse of wastes. In the present study, wastes derived from two different types of paper mill sludge (primary and secondary), was used for their bioconversion through the vermicomposting process using an indigenous species of earthworm (Perionyx excavatus). The maturity and stability stage of vermicompost production was assessed using FT-IR, GC-MS and TG analyses. During vermicomposting, different biochemical functional groups present in the wastes have shown differential chemical alteration and turnover as revealed by FT-IR spectroscopy. This study has also confirmed the trend of biodegradation of complex substances like lignin, cellulose, proteins etc. and thereby demonstrates the extent of mineralization. TG spectral analysis had revealed a mass loss of 80% and 71% in vermicompost produced from primary and secondary sludge respectively. GC-MS studies have also shown the presence of several humic acids like octadecanoic acid, heptadecanoic acid etc. in the decomposing substances demonstrating as an indicator of the maturity of products. This was further confirmed by the decrease of humification index which focuses the combined action of both earthworms and microbes in the degradation of organic wastes. The present study has highlighted the role of an indigenous earthworm in converting specific industrial wastes especially by recording the point of maturation using humic acids an indicator of the quality of decomposing of wastes following several instrumental applications.

Introduction: The earthworm coelomic fluid (ECF) has shown proven antiproliferative effect against breast, liver, gastrointestinal, and brain cancer, but it is least explored in oral cancer. The present in vitro study is an attempt to investigate the antiproliferative activity of ECF on oral cancer cell line squamous cell carcinoma (SCC)-9. Materials and Methods: ECF was collected from the species Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) stored at −80°C. Percentage inhibition of ECF on squamous cell carcinoma-9 cells in vitro was recorded at 24 h. Protein estimation was done using Bradford protein assay validated by the biuret method. Cytotoxicity was tested at 2.5, 5, 10, 20, 40, and 80 μg/ml concentrations by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay in SCC-9 cells in vitro. GraphPad Prism 7.0 software was used to calculate the inhibitory concentration (IC50). Chi-square test was used to analyze the difference between samples. Results: The test samples EE, EF, and PE inhibited the growth of SCC-9 cells significantly in a dose-dependent manner, and the IC50 values were found to be 4.6, 44.69, and 5.27 μg/ml, respectively. The antiproliferative effect was found to be variable among the three earthworm species with EE showing the most promising effect followed by PE and EF. Conclusion: Establishing the antiproliferative effect of ECF on oral cancer cells could be an initial step toward drug development and future anticancer research. The preliminary investigation has shown that ECF has a promising antiproliferative effect on oral cancer cells in vitro. SUMMARY The present pilot study evaluated the in vitro antiproliferative effect of earthworm coelomic fluid (ECF) of Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) on squamous cell carcinoma-9 cell line. The ECF inhibitory activity was promising at inhibitory concentration values of 4.6, 44.69, and 5.27 μg/ml, respectively. Further studies pertaining to antiproliferative mechanism of EE, EF, and PE have been planned. Abbreviations Used: ECF: Earthworm coelomic fluid, EE: Eudrilus eugeniae, EF: Eisenia foetida, PE: Perionyx excavatus, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, SCC: Squamous cell carcinoma, BSA: Bovine serum albumin, PBS: Phosphored buffered saline, ATCC: American Type Culture Collection

We aimed to recycle the agroindustrial waste resources - pressmud(PM), sugarcane trash (ST) and biomethanated distillery effluent(BE) and produced agronomic valid vermicompost using earthworm, Perionyx excavatus for maintaining natural soil organic and sustainable agricultural activity. Therefore, a series of studies were carried out to convert PM-ST-BE in different proportion vermibeds (T1-1000g PM+0g ST+790ml BE), (T2- 900g PM+100g ST+740ml BE), (T3-800g PM+200g ST+696ml BE), (T4-700g PM +300g ST+655ml BE), (T5-600g PM+400g ST+625ml BE) and (T6-500g PM+500g ST+542ml BE) into vermicompost. The study was to examine the activity of earthworm- growth, reproduction, vermicompost recovery and its nutrient status. The pronounced and better worm activity was found in all vermibeds, especially more in T1 and T3 vermibeds followed by others. This seems to be due to rich cellulose, OC, N, P, microbial activity, enhanced water holding capacity and palatability of the substrates. Enhanced microbial activity, humic acid content, NPK, normalized pH, declined OC, C-N, and C-P ratio, lignin, cellulose, hemicellulose and phenol in vermicompost than normal compost and control. The increased microbial-enzymatic activities contribute an increase in nutrients through nitrification, phosphate solubilization and mineralization. Reduction of OC, C-N, C-P ratio, lignin, cellulose, hemicellulose and phenol in the vermicompost are due to combined action of gut microflaura and earthworm during the vermicomposting process and utilization of these contents by the worm for their growth and reproduction. Finally, our study recommended for the production and application of vermifertilizer from lignocellulosic wastes using vermitechnology for sustainable activity.

Vermicomposting is a dependable waste recycling technology which greatly augments N and P levels mainly through microbial action. This paper aims to identify efficient N-fixing (NFB) and P-solubilizing (PSB) bacteria from earthworm intestines. Various combinations of vegetable market waste, rice straw, and cowdung were fed to two earthworm species (Eisenia fetida and Perionyx excavatus). Total organic C decreased, pH shifted towards neutrality, and NPK availability, and microbial (NFB, PSB, and total bacteria) population increased remarkably during vermicomposting with E. fetida. Therefore, 45 NFB and 34 PSB strains isolated from Eisenia gut were initially screened, their inter-dominance assessed, and 8 prolific strains were identified through 16SrRNA sequencing. Interestingly, two novel N-fixing strains of Kluyvera ascorbata emerged as an efficient biofertilizer candidate. Moreover, both N-fixing and P-solubilizing strains of Serratia and Bacillus were isolated from earthworm gut. All the isolated strains significantly improved soil health and facilitated crop growth as compared to commercial biofertilizers.

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.

Eisenia foetida and Perionyx excavatus are potent vermicomposting earthworms having immense importance in organic matter recycling under tropical conditions, particularly in India. Comparative assessment of the cultivable gut microbiome of these two epigeic earthworms after growth on lignocellulosic biomass, revealed populations of 3.2-8.3 × 10(9) CFU. Diversity analyses using 16S rDNA sequences revealed that the major dominating classes were Firmicutes (50-60%), followed by Actinobacteria (26.7-33%), and Alphaproteobacteria (5.6-6.7%). Despite exhibiting similar diversity indices and species richness, Betaproteobacteria (6.7%) and Gammaproteobacteria (11.1%) were solely present in E. foetida and P. excavatus, respectively. A set of 33 distinct morphotypes, including 18 from E. foetida and 15 from P. excavatus were selected. Carbohydrate utilization profiles generated using Hi-Carbo™ kits revealed that the isolates from the gut of P. excavatus - Arthrobacter pascens IARI-L13 and Bacillus subtilis IARIC were able to utilize 54 and 51.4% of the carbohydrates tested. Sorbose was not utilized, while unusual carbohydrates - adonitol and methyl-d-mannoside were utilized only by members from the gut of P. excavatus, while melizitose was utilized by those uniquely by E. foetida microbiome. Functional characterization revealed that β-glucosidase activity was most prevalent in the culturable microbial community. Alkaline and acid phosphatase activity was more widespread in the E. foetida gut microbiome. All the culturable gut bacterial isolates produced ammonia, but IAA was detected only in five cultures. The unique functional attributes of the two culturable microbiomes, grown on a similar diet, reveals the significance of proper selection of earthworm substrate combinations for effective vermicomposting.

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.

The aim of this work was to study nutrient recovery and vermicompost production from livestock solid wastes of indigenous and exotic cow breeds with epigeic earthworms. Fourteen days pre-decomposed dung of Vechur native (CD1) and exotic Jersey (CD2) breeds were vermicomposted with Perionyx excavatus and Eudrilus eugeniae for 45 days (Cycle I) and 90 days (Cycle II) including respective controls without earthworms. Vermicomposts from CD1 and CD2 substrates during Cycle I and II showed increase in NPK, Ca and micro-nutrients (P 0.05); whereas, pH, total organic carbon, C/N and C/P ratios disclosed decrease (P 0.05) over initial levels for both the earthworms. E. eugeniae was found efficient in vermicomposting cattle solid wastes in 45 days and CD1 yielded nutrient rich vermicompost. The study concludes that Cycle I is suitable for nutrient recovery and vermicompost production, in addition to mass multiplication of earthworms in Cycle II.

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.

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.

Two new earthworm species, one of Glossodrilus Cognetti, 1905 and the other of Martiodrilus (Maipure) Righi, 1995, are described from material collected from the coffee-growing region located in central-western Colombia: Glossodrilus chaguala sp. nov., and Martiodrilus (Maipure) quimbayaensis sp. nov. New records for this region are also reported for Glossodrilus griseus, Glossodrilus lacteus, Glossodrilus panikita, Aptodrilus fuhrmanni, Periscolex columbianus, Periscolex coreguaje, Pontoscolex (Pontoscolex) corethrurus, Dichogaster (Diplothecodrilus) affinis, Dichogaster (Diplothecodrilus) bolaui, Dichogaster (Diplothecodrilus) saliens, Amynthas (Amynthas) gracilis, Amynthas minimus, and Perionyx excavatus.

The Green Revolution in India made the country self-sufficient in food grain production. However, indiscriminate use of chemical fertilizers and pesticides during last five decades has made the soil unproductive and caused environmental pollution too. Environmentalists are now planning for a second Green Revolution, where reduced use of chemical fertilizers, pesticides, by recovery of soil health to increase in food grain production through organic farming along with conservation of biodiversity, is in the focus. Present paper deals with adoption of earthworm technology which could be the best choice for second Green Revolution. Earthworm technology is of two types: in-soil and ex-soil. In-soil technology involves mass culture of suitable geophagous and phytogeophagous earthworm species and their direct inoculation in soil with proper organic input. Ex-soil technology, popularly known as vermicomposting, allows quick transformation of organic wastes into plant-nutrient-rich compost through activities of phytophagous earthworm species.

Agriculture is an important part of the economy of the undivided Midnapore district. Agricultural land is its asset and most importantly its means of sustenance as well as survival. Earthworms are invertebrates that play a key role in recycling organic matters in soils. Since the intestines of earthworms harbor wide ranges of microorganisms, enzymes, hormones etc., these half digested materials decompose rapidly and are transformed into a stabilized material called vermicompost which is very useful for increasing the soil fertility. One has to look for these characters before recommending any species for vermiculture. In the present study, Perionyx excavatus specimens were collected from the undivided Midnapore district and from the Earthworms gut, bacteria, fungus, actinobacteria, and yeast were isolated and identified using various morphological and biochemical tests. All the bacterial isolates were identified using morphological study, staining techniques, and different biochemical tests such as catalase test, KOH test, H2 SO4 test, Starch hydrolysis test, oxidase test, and sucrose hydrolysis test. All the fungal, actinobacteria, and yeast isolates were subjected to staining and morphological characterization (color and texture of fungal colony). Bacterial isolates of genus Bacillus sp., Staphylococcus sp., Enterococci, Micrococcus sp., Enterobacter sp., and Citrobacter sp. were identified. Among the fungal isolates Aspergilus sp., and P. boydii were identified. Streptomyces sp., Nocardia sp. among the actinobacteria and Candida sp. among yeast were also found to be present in earthworm gut and these might play an important role along with the earthworm to increase the quality and fertility of soil.

Present study revealed the presence of 16 earthworm species belonging to 11 genera and four families viz. Megascolecidae (Amynthus alexandri, Metaphire houlleti, Lampito mauritii, Kanchuria sp1, Perionyx excavatus), Octochaetidae (Eutyphoeus gigas, Eutyphoeus comillahnus, Eutyphoeus orientalis, Octochaetona beatrix, Dichogaster bolaui, Lennogaster chittagongensis, Lennogaster yeicus), Moniligastridae (Drawida papillifer papillifer, Drawida assamensis, Drawida nepalensis) and Glossoscolecidae (Pontoscolex corethrurus) in the soils of five bamboo species [Bambusa balcooa (Sil Barak), Melocanna baccifera (Muli), Bambusa polumorpha (Bari), Bambus cacharensis (Bom) and Bambus bambus (Katabarak)] of West-Tripura. While four earthworm species viz. Metaphire houlleti, Drawida assamensis, Drawida papillifer papillifer and Pontoscolex corethrurus were common to all species of bamboo plantations, the rest showed restricted distribution. Among the earthworm species 4 were exotic (Amynthus alexandri, Metaphire houlleti, Dichogaster bolaui and Pontoscolex corethrurus) and the others were native to the Indian sub-continent. In general, earthworms under the bamboo plantations occurred within temperature range of 21.6 °C–28.0 °C, pH 4.0–7.0, organic matter 0.56–5.99 %, moisture 9.6–31.7 %, water holding capacity 14.6–43.9 % and bulk density 0.7–1.8 g cm−3. The average density and biomass of the earthworms in the studied places were 108 ind m−2 and 44 g m−2 respectively. Earthworm diversity, dominance and evenness indices showed the values 1.00, 0.47 and 0.70 respectively. Earthworm density and biomass showed a negative correlation with temperature whereas those had a strong positive correlation with pH, moisture and organic matter of the soils.

The aim of this study was to investigate the biochemical responses in three earthworm species, Eudrilus eugeniae, Perionyx ceylanensis and Perionyx excavatus induced by exposure to pesticide (carbaryl) and metal (lead) in soil. Earthworms were exposed to increasing concentrations of carbaryl (12, 25 and 50 mg kg −1) and Pb (75, 150 and 300 mg kg −1) in the soil for different periods of time and the biochemical changes were determined. The protein content was slightly increased in E. eugeniae, P. ceylanensis when exposed to the lower concentration of Pb (75 mg kg−1). But decreased protein content was observed in P. excavatus at the higher concentration of carbaryl (50 mg kg−1), and with exposure to 300 mg kg−1 Pb in P. ceylanensis. The changes in the levels of antioxidant enzymes glutathione-S-transferase (GST), glutathione peroxidase (GPx), reduced glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) were used as functional biomarkers to determine the effect of pesticides and metals on earthworms. Among these, GST exhibited more sensitive response to both carbaryl and Pb as compared to the control. Other antioxidant enzymes showed little variation at the time of exposure. The results of our study suggested that both carbaryl and Pb induced the production of Reactive oxygen species (ROS) by causing oxidative damage to cells. Due to an adaptive response of earthworms in contaminated soils, they could be used as significant biomarkers to asses the toxicity in the environment of the soil ecosystem. This is the first report on the comparative study of antioxidant enzyme activities in three earthworm species under normal and stress induced conditions.

Diafenthiuron, an insecticide widely used in the management of pests of cardamom and cotton, was assessed for its toxicity-effect on beneficials commonly found in these ecosystems. Diafenthiuron was found to be toxic to honey bees, the prime pollinators of crop plants. Diafenthiuron at the highest tested dose caused 40% mortality to the coccinelid grubs at 48 h after treatment so, diafenthiuron was found to be slightly harmful. Monocrotophos, on the other hand has been found to be a highly toxic pesticide. Diafenthiuron is moderately harmful to the adults of Chelonus blackburni L. The testing was done using the insecticide diafenthiuron. An insecticidecoated vial (scintillation) bioassay was performed. It was found that there was 86.67% mortality in 48 h, at the recommended dose. Diafenthiuron is highly toxic to the silkworm, killing more than 80% of the caterpillars in 24 h, at all the doses tested. Diafenthiuron, even in the highest dose tested, is non-toxic to the earthworm, Perionyx excavatus, which was found to have a 3.33% mortality. As far as fish are concerned, the common carp, Cyprinus carpio L., is found to be highly susceptible to diafenthiuron and even doses 10 times lower than the field dose can kill the fish within 6 h.

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.