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

Purpose 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 earthwormâPerionyx excavatus (Perrier, 1872) and produce quality vermifertilizer. Methods 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 activityâgrowth, reproductive performance (cocoon production and hatchling number) and recovery of vermicompost was also studied. Results The obtained results clearly showed that vermicompost from CLL admixed with cowdung at 2:2 ratio had lower pH, organic carbon, CâN ratio, CâP 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. Conclusion Through vermitechnology way of producing agronomic valid vermicompost using natural waste resources like CLL and animal dungs can be used as bioorganic 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.

Acetamiprid is a broad-spectrum insecticide, belonging to the neonicotinoid compounds group, which has been extensively applied throughout the globe. Recently, indiscriminate use of these compounds was reported to cause fatal impacts on non-targeted soil organisms. Hence, the present study aimed to examine the impact of acetamiprid on Indian indigenous earthworm, Perionyx excavatus. Acute toxicity revealed an LC50 concentration of 0.25 µg/cm2 for filter paper test/72 h and 400 µg/kg for artificial soil test/14 days. Oxidative stress (ROS) and various biomarkers including superoxide dismutase, catalase, glutathione S-transferase, malondialdehyde content and DNA damage were measured. The results of the biomarker responses confirmed the acetamiprid exposure can cause toxicity to P. excavatus. In addition, cell density (20 × 102 cell mL/mg) and cell viability (40%) were significantly (p < 0.05) reduced. Further, the ecotoxicological assessment made through this study can be utilized as good evidence to toxicity of neonicotinoids to non-targeted indigenous organisms. [ABSTRACT FROM AUTHOR]

The ideal condition of earthworm gut promotes growth and multiplication of beneficial soil microorganisms eliminating pathogens and converts organic wastes into nutrients rich compost. The present study has been carried out to determine the population dynamics of earthworm gut bacteria and to find out relative abundance of different functional bacterial groups in the foregut, midgut, and hindgut of earthworm Perionyx excavatus. To assess bacterial diversity, a viable plate count method was adopted. In the different gut region of earthworm, aerobic heterotrophic, amylolytic, Bacillus, Gram-negative, proteolytic, fat hydrolyzing, nitrate-reducing, nitrifying, asymbiotic nitrogen-fixing, Azotobacter, and phosphate solubilizing bacterial populations ranged from 22.2 to 241.6 × 106, 8.0 to 171.60 × 106, 1.83 to 2.79 × 106, 10.68 to 23.04 × 104, 3.70 to 5.52 × 104, 59.60 to 208.40 × 104, 1.86 to 7.34 × 104, 10.94 to 19.78 × 104, 0.80 to 3.42 × 104, 7.83 to 13.70 × 104, 1.31 to 2.67 × 104 cfu/ml gut suspension, respectively. The results of the one-way ANOVA revealed that the bacterial load of most of the bacterial groups was significantly higher (p < 0.05) in the hindgut region, followed by midgut and foregut. Only the density of the proteolytic group was significantly higher (p < 0.05) in the midgut region followed by foregut and hindgut. Starch hydrolyzing bacteria constitute the largest group of bacteria in the gut content. From principal component analysis, two components were extracted with the eigenvalues of 8.485 and 1.132. Agglomerative hierarchical cluster analysis revealed that the bacterial populations were clustered into four different groups. Quantitative variation among bacterial groups in earthworm's gut seems to determine the soil health and composting efficiency; from this point of view, the present study will provide a better understanding about different functional bacterial groups of earthworm's guts and might be helpful in sustainable agriculture and waste management. [ABSTRACT FROM AUTHOR]

[Display omitted] • A total of 55 known and 50 novel miRNAs are identified in the earthworm. • pex-miR-125, pex-miR-71, pex-miR-365, and pex-miR-36 are up-regulated during regeneration. • Utilizing miRanda tool, 50 novel miRNAs are predicted from 1938 mRNA targets. • Key regulatory genes Hox, MAPK, Wnt, SoxB, Notch, MYC are involved in regeneration. Earthworm, P. excavatus, is an ideal model organism for studying regeneration. Due to its prodigious regeneration capability, the amputated head part of the earthworm can regenerate completely within 22 days. MicroRNAs (miRNAs) regulate specific genes and are involved in essential biological processes, including regeneration. In this study, we conducted a comprehensive analysis of miRNA profiling of the earthworm, P. excavatus, during the process of anterior regeneration. Our investigation involved in the identification of 55 miRNAs from 30 distinct miRNA families that exhibit significant relevance to wound healing and regeneration. Notably, we have identified 50 novel miRNAs and predicted their pre-miRNA secondary structures using MIREAP. Both Known and Novel miRNAs are validated using qPCR. In addition, we employed the miRanda algorithm to predict the interactions between these miRNAs and their target mRNA transcripts. Based on the miRanda target prediction results, we identified the target genes such as Wnt , Myc , MAPK , SoxB , IHH , Hox , and Notch. These findings indicate that the potential targets of these miRNAs might play crucial roles in various functions related to wound healing, tissue restoration, and regeneration. Furthermore, the acquisition of these findings provides a unique perspective on understanding the molecular mechanisms driving epimorphosis regeneration in connection with miRNAs for the development of miRNA-based therapeutics. [ABSTRACT FROM AUTHOR]

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]

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.

Alternative plant growing inputs have been increasingly sought out to reduce the economic-environmental impacts of intensive use of synthetic fertilizers in conventional agriculture. This study aimed to evaluate the production components of lettuce in response to the use of earthworm compost in substrate compositions. The experiment was carried out in a greenhouse using a completely randomized design in a 3 x 5 factorial scheme, with six replications, testing three earthworm species (Eisenia andrei, Eudrilus eugeniae, and Perionyx excavatus) and five drilocompost ratios (0%, 25%, 50%, 75%, and 100%), produced by these species and added to the composition of substrates by mixing them with soil. Production components evaluated comprised total and commercial numbers of leaves, commercial fresh weight, as well as shoot and total fresh and dry weights. Lettuce production components increased as the drilocompost ratio in substrates was raised, regardless of the earthworm species. Moreover, the addition of 25% this drilocomposted material in the dystrophic soil was enough to improve its fertility condition. In general, substrates prepared with worm castings from Perionyx excavatus and Eudrilus eugeniae were more efficient in increasing lettuce production components than that from Eisenia andrei. [ABSTRACT FROM AUTHOR]

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<0.05) higher in cow dung (maximum weight mg worm-1 629 and growth rate mg worm-1 day-1 22.91). The rate of reproduction (0.3 cocoons worm-1 day-1 and 3.31 juveniles adult-1 week-1) was significantly higher (P<0.05) in acacia-cow dung mixture. The cocoon production was maximum on the 30th day in all experimental diets (cow dung, acacia-cow dung, bamboo-cow dung, mikania-cow dung). All the diets showed a maximum peak of juvenile production on the 45th day. The lowest rate of biomass increase as well as the rate of reproduction were observed in mikania-cow dung mixture. Present result indicates that cow dung and acacia leaf litter can be used as vermiculture substrate for P. excavatus. [ABSTRACT FROM AUTHOR]

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 treatment (T1), cow dung was used as the only food source for the earthworm. In another five treatments, 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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

• Seaweeds in combination with cowdung (1:1) were subjected to vermicomposting for 60 days. • Earthworms significantly increased the EC and nutrients-NPK and reduced total organic carbon and C/N. • Vermicomposting by Perionyx excavatus increased the total microbial population. • Growth and reproduction of the earthworms shows the acceptability of seaweed substrates. 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 < 0.05) higher than the substrates without earthworms after 60 days. The physico-chemical characteristics of the vermicompost showed increased levels of electrical conductivity, NPK, while organic carbon, C:N ratio and organic matter content showed reduction in all seaweed + CD substrates. A higher amount of NPK of 1.38, 0.69 and 2.41% respectively in the vermicompost of Turbinaria ornata + CD was recorded with the respective percentage increase of 50.00, 68.29, and 34.64 over compost, and the values were significantly higher (P < 0.05) than in the initial substrate as well as the worm-unworked compost of Turbinaria ornata + CD. The activity of earthworms during vermicomposting of seaweed substrates significantly increased the total microbial population. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Vermicomposting of seaweeds + cowdung in 1:1 ratio was studied. • NPK showed significant increase while organic carbon and C/N showed reduction. • Total microbial population invariably increased by the action of earthworms. • Earthworms strongly accelerated vermiconversion of seaweed + cowdung substrates. • Growth and reproduction of earthworms showed acceptability of seaweed substrates. Abstract 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. [ABSTRACT FROM AUTHOR]

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

Earthworm, Perionyx Excavatus, has been studied and the potential of this epigeic earthworm species for processing organic waste is well known. The substrate and feeding material were considered as effector of the growth, life cycle of earthworms. The current study carried on the evaluation of various cultivating and feeding materials that influenced on growth of earthworms, by calculating biomass, the total number of worms and relative growth rate. As the results, the cow manure fed in cultured and feeding materials showed the suitable in the growth of earthworm, with significant differences from other group which based on worm manure as the substrate and feeding materials. For further study, the adding component, including coir, straw and soil have to be studied to find out the better materials in earthworm cultivation.

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.

Graphical abstract Highlights • Concentrations of heavy metals were significantly reduced in paper mill sludge by earthworm. • Heavy metals were accumulated by earthworm internal body. • High concentration of paper mill sludge showed the highest incidence of damaged tissues in earthworms. • Vermistabilization is a low-cost eco-friendly technology to combat land pollution. Abstract 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. [ABSTRACT FROM AUTHOR]

OBJECTIVE: To investigate SOD-liked, GPx-liked and cellular anti-oxidation activities of fractionated mucus proteins from Perionyx excavatus (Pe) and Eudrilus eugeniae (Ee) in the MC3T3 osteoblast precursor cell line. MATERIALS AND METHODS: The crude mucus proteins were extracted from Pe and Ee and fractionated using the anion exchange chromatography method with FPLC. The fractionated proteins were studied to determine their SOD-liked and GPx-liked activities. The most efficient fractions were studied for cellular anti-oxidation activities in the MC3T3 cell line including scavenging, protecting and repairing conditions. RESULTS: The results showed that the highest SOD-liked activities of EeANX1 were found at 4.17 µg/ml and GPx-liked activities of EeANX4 were found at 5.26 µg/mL. EeANX1 had no cytotoxic effect on MC3T3 at 500 µg/mL, and the IC50 of EeANX4 was over 300 µg/mL. When both EeANX1 and EeANX4 were investigated for cellular anti-oxidation activity in MC3T3 cells at 20 µg/mL, the cellular superoxide and total ROS production of were significantly (p<0.05) lower than those of 250 μM pyocyanin (ROS inducer) in ROS scavenging, protecting and repairing conditions. The 20 µg/mL of EeANX1 and EeANX4 demonstrated increased SOD and GPx activities in MC3T3. Investigation of the scavenging, protection and repairing conditions of MC3T3 treated with 20 µg/mL each of EaANX1, EaANX4 and 250 μM pyocyanin demonstrated that the proteins had significantly lower SOD activities and higher GPx activities than the 250 μM pyocyanin. CONCLUSIONS: The EeANX1 and EeANX4 fractions demonstrate SOD-liked and GPx-liked activities, as well as cellular anti-oxidation activities. These fractions could be developed as a natural anti-oxidant. This research could provide benefit to the study of cellular anticancer. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The feasibility of vermicomposting water hyacinth (WH) [Eichhornia crassipes (Mart.) Solms] mixed with pig manure (PM) in different proportions was tested using tropical composting earthworm Perionyx excavatus. Earthworms grew and reproduced normally until the incorporation of 50% WH in initial substrate. Higher water hyacinth proportions induced earthworms' mortality and significantly affected the numbers of hatchlings and cocoons produced during vermicomposting period. The influence of the application of compost/vermicompost obtained from water hyacinth mixed with pig manure was also studied on seeds germination. Only water hyacinth substrate with 25% WH + 75% PM enhanced seeds germination for Oryza sp. and Nasturtium officinale. At the end of experiments, a significant decrease was observed in organic carbon content for each tested substrates (S1 to S8), in total nitrogen (N) for substrates containing 70% to 100% of water hyacinth (S5 to S3) and compost substrates (S1 and S2). An important decrease was also noted in total potassium for all vermicompost substrates (S3 to S8), in total magnesium for composted substrates (S1 and S2), and in C/N ratio for substrates containing 0% to 50% of water hyacinth (S8 to S6). Whereas total N in vermicompost containing 0% to 50% of water hyacinth (S8 to S6), total phosphorus, total potassium in composted substrates (S1 and S2), total magnesium in vermicompost substrates (S3 to S8) and C:N ratio in substrates containing 70% to 100% of water hyacinth (S5 to S3) expressed a significant increase after eight weeks. The result suggested that water hyacinth could be potentially useful as raw material in vermicomposting and biofertilizing if mixed with 75% of pig manure.

OBJECTIVE: This study aimed to investigate the enzymatic anti-oxidative, hemolytic and cytotoxic activities of crude mucus proteins from the two earthworms including Eudrilus eugeniae (African night crawler) and Perionyx excavatus (Blue worm). MATERIALS AND METHODS: The bioactivities were determined by hemolytic activity, cytotoxic activity using HepG2 human hepatocellular carcinoma cells, L-929 mouse fibroblast and TK6 human lymphoblast cell lines and antioxidant activity. RESULTS: The results indicated that the hemolytic activity of mucus proteins of P. excavatus was higher than that of E. eugeniae.The cytotoxic activity of the mucus proteins of E. eugeniae could inhibit the growth of HepG2 with the IC50 at 144.2 ± 0.18 μg/mL but showed no effect on L-929 and TK6. On the other hand, the crude proteins of P. excavatus decreased cell viability of both L-929 and HepG2 with their IC50 respectively were 6.87 ± 0.16 and 174.3 ± 0.19 μg/mL but they did not reduce the growth of TK6 cell line. The SOD-like activity of P. excavatus and E. eugeniae crude proteins were found with the IC50 at 149 μg/mL and 386.2 μg/mL, respectively. For GPx-like activity, crude proteins of P. excavatus exhibited significant (p ≤ 0.05) greater activity than those of E. eugeniae when tested at the concentration 100 μg/mL. CONCLUSIONS: This study revealed the bioactivities of crude mucus of earthworms which served as alternative natural proteins for the prophylaxis or treatment of free radical-related diseases as well as development of dietary supplements and cosmetics. [ABSTRACT FROM AUTHOR]

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 1 m 3 m −2 d −1 (VF1) and 1.5 m 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 20 days in VF1 and 14 days 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. [ABSTRACT FROM AUTHOR]

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 CO 2 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. [ABSTRACT FROM AUTHOR]

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 × 109 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. [ABSTRACT FROM AUTHOR]

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, H2SO4 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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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 (LC50 mix 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. [ABSTRACT FROM AUTHOR]