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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.

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).

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.

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.

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.

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

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.

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.

Methyl tert-butyl ether (MTBE) and tert-butyl alcohol (TBA) are among the major soil contaminants that threaten the health of soil ecosystems. Many MTBE-contaminated sites accumulate TBA, because TBA is the intermediate of MTBE biodegradation. To access the risk of MTBE and TBA in soil, we investigated the combined toxicities of MTBE and TBA using two earthworm species, Perionyx excavatus and Eisenia andrei, as well as the toxic effects via different exposure routes. The combined toxicity showed weak antagonistic effects (LC50mix values were slightly greater than 1.0), and sensitivity toward same pollutants differed in the two earthworm species. Moreover, the toxicity of MTBE and TBA was also affected by the exposure route; both filter paper and artificial soil tests showed that dermal-only exposure to MTBE had an even greater toxic effect than combined dermal and oral exposure. Thus, we suggest that diverse environmental factors including organic materials, the physicochemical properties of the contact media, and the exposure routes of the organism, should be taken into consideration when assessing the effects of pollutants on organisms in diverse environmental systems.

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.

In the present study, Eudrilus eugeniae and Perionyx excavatus were used for vermistabilization of textile mill sludge in different combinations with cowdung for 60 days. A higher percentage of metal removal was observed in earthworm treated mixtures for cadmium (54.5%) followed by copper (36.0%), chromium (37.0%) and zinc (35.9%). Vermistabilized textile mill sludge + cowdung (1:1) showed a maximum percentage increase in total NPK, a significant (P

This study describes a new and sensitive method for measuring the in vivo and in vitro cytotoxicity of 2 earthworm species, Eisenia andrei and Perionyx excavatus, exposed to copper. Specifically, we measured the number of coelomocyte cells that were affected by copper following in vivo and in vitro exposure by flow cytometry, after calcein acetoxymethyl ester (calcein-AM) staining. We found that the coelomocyte viability of both earthworm species was noticeably reduced in the in vivo cytotoxicity test at concentrations of 100mg/kg copper in dry soil. However, pathological symptoms, such as mucous secretion and bleeding, swelling, thinning, and fragmentation, and burrowing symptoms were not evident following exposure to copper levels of

A survey to know the diversity of endoparasitic protozoan parasites of the earthworms of West Bengal, revealed a new species of the genus Monocystis Stein, 1848, that resides in the seminal vesicle of the earthworm Perionyx excavatus Perrier. Monocystis kuidongae sp. nov. is characterized by ovoidal gamonts (trophozoites) with inconspicuous mucron and measure 69.53–102.25 (86.29 ± 11.48) μm × 24.54–61.35 (35.17 ± 12.82) μm. Shape of the gametocysts are almost rounded and measure 81.80–110.43 (96.11 ± 8.85) μm in diameter. Oocysts are navicular with pointed ends and measure 11.55–12.32 (11.78 ± 0.36) µm × 5.00–5.77 (5.39 ± 0.25) µm.

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.

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.

The earthworm provides an excellent model for investigating regeneration. Here we report the full-length cloning of three labial genes (Pex-lab01, Pex-lab02, and Pex-lab03) in the earthworm Perionyx excavatus. To analyze their expression pattern during head and tail regeneration, we used the reverse transcription-polymerase chain reaction. Our results indicate that the three labial genes were expressed only in the head-regenerating tissues. Also, we found that the expression of Pex-lab01 and Pex-lab02 is up-regulated, and this indicates their involvement in wound healing and the blastema formation processes during early head regeneration.

Three different earthworm species Eisenia fetida, Eudrilus eugeniae and Perionyx excavatus in individual (Monocultures) and combinations (Polycultures) were utilized to compare the suitability of worm species for vermicomposting of filter mud as well as the quality of the end product. The filter mud blended with saw dust can be directly converted into good quality fertilizer (vermicompost). Eight different reactors including three monocultures and four polycultures of E. fetida, E. eugeniae and P. excavatus and one control were used for the experiment. Vermicomposting resulted in significant reduction in C/N ratio, pH, total organic matter (TOC) but increase in electrical conductivity (EC), total nitrogen (TN), total phosphorus (TP) and macronutrients (K, Ca and Na). Oxygen uptake rate (OUR) dropped up to 1.64-1.95 mg/g (volatile solids) VS/day for monoculture reactors and 1.45-1.78 mg/g VS/day for polycultures reactors, respectively, after 45 days of vermicomposting. Cocoon production and the earthworm biomass increased as vermicomposting progressed. On an overall the mono as well as polyculture reactors produced high quality stable compost free from pathogens and no specific differentiation could be inferred between the reactors.

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.

The present paper is aimed at safe reuse and recycling of sewage sludge (SS) and production of good quality compost using vermicomposting. Three different earthworm species Eiseniafetida (E. fetida), Eudrilus eugeniae (E. eugeniae), Perionyx excavatus (P. excavatus) in individual and combinations were utilized to compare the suitability of worm species for composting of sewage sludge as well as the quality of the end product. The sewage sludge without blending can be directly converted into good quality fertilizer (vermicompost). Vermicomposting resulted in reduction in C/N ratio 25.6 to 6-9, TOC (25%) but increase in electrical conductivity (EC) (47-51%), total nitrogen (TN) (2.4-2.8 times), potassium (45-71%), calcium (49-62%), sodium (62-82%) and total phosphorous (TP) (1.5-1.8 times), which indicated that sewage sludge can be recycled as a good quality fertilizer. The present study also inferred that the application of sewage sludge in the agricultural fields after vermicomposting would not have any adverse effect as the heavy metals (Cu, Mn, Pb and Zn) are now within the permissible limits.

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.

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.

Recycling of guar gum industrial waste through vermitechnology was studied under laboratory conditions by using composting earthworm Perionyx excavatus (Perrier). Three different combination of guar gum industrial waste namely guar gum industrial waste:cow dung:saw dust in 40:30:30 ratio (T1), guar gum industrial waste:cow dung:saw dust in 60:20:20 ratio (T2), and guar gum industrial waste:cow dung:saw dust in 75:15:10 ratio (T3) were used for vermicomposting experiments. Chemical changes during vermicomposting were measured and comparatively T2 showed great increase (from its initial level) for total N (25.4%), phosphorus (72.8%) and potassium (20.9%) than the other treatments. T2 also showed higher vermicomposting coefficient (VC), higher mean biomass for P. excavatus (146.68 mg) and higher cocoon production (about 21.9% and 645.5% more than the T1 and T3, respectively). Maximum earthworm mortality during vermicomposting was recorded with T3 treatment while zero mortality was recorded for T2 treatment after 150 days. Overall, T2 treatment appeared to be an ideal combination for enhancing maximum biopotential of earthworms to management guar gum industrial waste as well as for earthworm biomass and cocoon production.

To establish the use of Metaphire posthuma as a sensitive model for ecotoxicological studies, the comparative effects of five metals on the hatching profiles of the cocoons of the earthworms, Metaphire posthuma, Eisenia foetida and Perionyx excavatus, were studied. The cocoons of the three species of earthworms were exposed to copper, chromium (III), chromium (VI), lead and zinc at 1.25, 2.5, 5.0 and 10.0ppm. Viable cocoons were incubated at 20 ± 1°C by using the immersion method. The results indicated that the inhibition of cocoon hatching was concentration dependent. The normal hatching, delayed hatching and non-viability of cocoons were recorded. At a concentration of 1.25ppm, there was almost no effect on the hatching of the cocoons of all three species of earthworms, except when exposed to chromium (VI), but higher concentrations (2.5 and 5.0ppm) caused severe effects. It was concluded that M. posthuma was more sensitive than the other two species, and that it is a suitable model for use in eco-toxicity testing.

An earthworm bioassay was conducted to assess ecotoxicity in methyl tert-butyl ether (MTBE)-amended soils. Ecotoxicity of MTBE to earthworms was evaluated by a paper contact method, natural field soil test, and an OECD artificial soil test. All tests were conducted in closed systems to prevent volatilization of MTBE out of test units. Test earthworm species were Perionyx excavatus and Eisenia andrei. Mortality and abnormal morphology of earthworms exposed to different concentrations of MTBE were examined. MTBE was toxic to both earthworm species and the severity of response increased with increasing MTBE concentrations. Perionyx excavatus was more sensitive to MTBE than Eisenia andrei in filter papers and two different types of soils. MTBE toxicity was more severe in OECD artificial soils than in field soils, possibly due to the burrowing behavior of earthworms into artificial soils. The present study demonstrated that ecotoxicity of volatile organic compounds such as MTBE can be assessed using an earthworm bioassay in closed soil microcosm with short-term exposure duration.

The performance of four species of earthworm––Eudrilus eugeniae, Kinberg, Drawida willsi Michaelsen, Lampito mauritii, Kinberg and Perionyx excavatus, Perrier––born and grown in vermireactors fed with paper waste was studied over six months, in terms of vermicast output per unit feed, production of offspring, and increase in worm zoomass. These were compared with the performance of the previous generation which had been raised to adulthood on cowdung as principal feed before shifting them to vermireactors operating on cowdung-spiked paper waste. The results indicated that except with D. willsi of which the second generation performed only a shade better than the first, there was significant improvement in vermicast output, animal growth, and reproduction in the second generation compared to the first. The results indicated that cowdung-spiked paper waste can be an adequate food for successive generations of earthworms and that reactors can be operated indefinitely on this feed. The results also indicated that the earthworm generations born and raised in vermireactors operated on this feed become better vermiconverters of this feed than the parent earthworms.

An experiment was conducted during 1998-1999, in a deciduous forest located in the semi-arid tropics of central India, to evaluate the suitability of different forest litters as food material for the tropical epigeic earthworms i.e. Eisenia fetida (Savigny), Perionyx excavatus (Perrier) and Dicogaster bolaui (michaelsen). The aim was to examine the influence of these earthworms on the decomposition processes of three types of forest litters i.e. Tectona grandis (teak), Madhuca indica (mahua) and Butea monosperma (palas), on the maintenance of quality in a vermicomposting system, and to assess the effect of applications of in situ prepared vermicomposts on the growth of forest trees. The results indicated that T. grandis litter was the most suitable food material for the earthworms possibly because it contained high reserves of mineral nutrients. Comparisons of the survival and reproduction rates of the three epigeic earthworm species indicated that a higher reproduction rate was maintained for E. fetida compared to P. excavatus and D. bolaui in the decomposition of these forest litters. The rates of growth and population increases of E. fetida approximately doubled after 12 weeks of litter decomposition. The litter decomposition process was associated strongly with the quality of the materials and their chemical composition. Irrespective of earthworm inoculations, the levels of available nutrient such as NH(4)-N, NO(3)-N, available P and K increased significantly (p/=0.05) in the order T. grandis litter compostM. indica litter compostB. monosperma litter compost. The mature decomposed litter had lower C/N ratios (11.3-24.8:1), water-soluble carbon (0.30-0.58%), water-soluble carbohydrates (0.35-0.71%) and larger cation exchange capacity/total organic carbon ratios than the values in the parent forest litter. The lignin content increased with maturation with a concomitant decrease in cellulose resulting in higher lignin/cellulose ratios. Application of all three vermicomposts to forest trees significantly improved their heights and diameters over those of control trees, although the increases were lower than those resulting from the chemical fertilizer applications. However, soil biological activities i.e. soil respiration, soil microbial biomass carbon and dehydrogenase activity were greater by application of vermicomposts over that after application of inorganic fertilizer in a new plantation of T. grandis.

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.

The recently developed concept of high rate vermicomposting was successfully used to enable direct vermicomoposting of neem leaves—without any pre-composting or cow dung supplementation as previously reported processes had necessitated. All the three epigeic species of earthworms that were explored, Eudrilus eugeniae, Eisenia fetida and Perionyx excavatus, provided efficient vermicast production with no mortality, persistent gain in body mass and good fecundity over the 16 months long period of reactor operation. In this period, all reactors were pulse-fed at the solid retention time of 20 days and were operated in the pseudo discretized continuous operation protocol developed earlier by the authors. With this, it was possible to almost completely dampen the influence of natural biodegradation of the feed or grazing by the earthworm born in the vermireactors. The findings, thus, conclusively prove that, all-through, the brisk vermicomposting was caused almost entirely by the action of the ‘parent’ earthworms on fresh feed.