Australian Journal of Basic and Applied Sciences, 3(4): 3671-3676, 2009 ISSN 1991-8178 2009, INSInet Publication

Chemical Analysis of Vermicomposts / Vermiwash of Different Combinations of Animal, Agro and Kitchen Wastes
1

Gorakh Nath, 2 Keshav Singh and 2 D.K .Singh

PhD Scholar Department of Zoology, D.D.U. Gorakhpur University, Gorakhpur -273009, U.P., INDIA 2 Department of Zoology, D.D.U. Gorakhpur University, Gorakhpur -273009, U.P., INDIA

Abstract: In India million tons of livestock excreta, agro and kitchen wastes are produced every year which are serious problems for society. This work to evaluate the potential of an epigeic earthworm Eisenia foetida to convert the different combination of variety of wastes in to rich nutrient vermicomposts/vermiwash and pre and post chemical analysis of feed mixtures. Vermicomposting results in significant decreased in pH, Total organic carbon (TOC), electrical conductivity (EC) and C:N ratio while significant increase in total Kjeldohl nitrogen (TKN) available phosphorus, exchangeable potassium and calcium in vermicomposts/vermiwash. The increased level of plant n utrients in final p rod ucts in different organic resources dem onstrated that the vermicompost/vermiwash of these wastes will be a valuable biofertilizer for sustainable land restoration practices. This study clearly indicates that vermicomposting of animal, agro/kitchen wastes not only produced a valuable vermicompost/vermiwash but also increased level of plant growth supplements in final vermicompost. Key words: W astes, Eisenia foetida, Vermicomposting, vermiwash, Chemical analysis INTRODUCTION The excess uses of chemical fertilizers and pesticides have made our soil sick and problematic and cause environmental hazards which affect the human health and environment. Million of tons of animal, agro and kitchen wastes are produced annually and have odor and pollution problems (Gupta, 2005; Reinecke et al., 1992; Garg et al., 2006). M uch attention has been paid in recent passed years to manage different organic wastes resources at low input as well eco-friendly basis. Vermicomposting, through earthworms, is an ecobiotechnological process that transforms energy rich and complex organic substances in to a stabilized vermicomposts (Bentize et al., 2000). Vermicomposting is stabilization of organic material involving in the joint action of earthworms and micro organisms. Although microbes are responsible for the biochemical degradation of organic matter, earthworms are the important derivers of the process, conditioning the substrate and altering the biological activity (Aira et al., 2002).The epigeic earthworm were utilize for organic wastes management (Suthar, 2006; Benitez et al., 2005; Garg and Kaushik, 2005; Benitez et al.,2005). The epigeic earthworm were utilize for organic wastes management (Suthar, 2006; Garg and Kaushik, 2005; Benitez et al., 2005). During vermicomposting the nutrients are released and converted into soluble and available forms to plants (Ndegwa and Thompson, 2001). The vermicomposting through different species of earthworm has been studied (Edwards et al., 1998; Kale et al., 1982). The earthworm Eisenia andrei converted paper-pulp mill sludge mixed with primary sludge in to the vermicompost ( Elvira et al.,1996). Suthar (2008) studied that the post harvest crop residues and cattle shed manure were recycle through vermicomposting by using the epigeic earthworm Eudrilus eugeniae. It has been well established that epigeic forms of earthworms can hasten the composting process to a significant extent with production of better availability of vermicomposts (Ndegwa and Thomson, 2001). The epigeic earthworm E. foetida is a suitable species for management of wastes which are utilized successfully in vermicomposting (Gunadi and Edwards, 2003; Chaudhari and Battacharjee, 2002; Garg et al., 2003; 2004). The aim of present study to explore the suitability and potential use of Eisenia foetida to management of animal, agro and kitchen wastes with the production of vermicompost/vermiwash.

Corresponding Author: Dr. Keshav Singh, Department of Zoology, D.D.U. Gorakhpur University, Gorakhpur 273009, U.P., INDIA Phone no. 09450433313 Email: [email protected] 3671

Aust. J. Basic & Appl. Sci., 3(4): 3671-3676, 2009 M ATERIALS AND M ETHODS Collection of Wastes: Animal wastes (cow, buffalo, sheep, horse, goat dung) were collected from different cattle farms and different agro/ kitchen wastes were collected from rural and urban parts of Gorakhpur district. After the collection, the different combinations of organic wastes sprayed in a layer of 1-2 feet and exposed to sun light for 5 to 10 days to removing the various harmful organisms and noxious gases (Bhatnagar and Palta 1996). Collection of Earth Worm : Earth worms Eisenia foetida were collected from U.P.agro, states industrial area, Gorakhnath, Gorakhpur. The collected earthworms acclimatized in the laboratory conditions. Experim ental Design for Verm icom posting: The experiment for vermicomposting was conducted on cemented earth surface. For the vermibed of each different combination of animal, agro/ kitchen wastes in 1:1 ratio, in the size of 3m x 1m x 9cm. After this moistened it and inoculated at least 2kg of cultured Eisenia foetida in each vermibed, then covered the bed by jute pockets. Moistened the vermibed daily up to 40 to 45 days for maintaining of moisture. After one week interval turned the mixture of bed manually up to 3 weeks. After 50 to 60 days granular tea like vermicompost appear on the upper surface of beds. Extraction of Verm iwash: Vermiwash extracted through vermiwash collecting device which is made up of plastic or metals drum having capacity of 5 liter and a tap at the bottom the drum filled with broken breaks, about 10cm thickened which is followed by sand layer of 2-3cm thickness lastly with filled with vermicompost with heavy population of earth worms simultaneously added fresh water in to drum and a container kept bellow the tap of drum. The watery yellowish to black extract of vermicompost, vermiwash drainage out off drum. After 1to 2 days the process of extraction has been completed. The different concentrations of collected vermiwash were used for chemical analysis. Chem ical Analysis: The chemical analysis of raw organic wastes and final vermicompost/ vermiwash of different wastes were observed. Total organic carbon (TOC) was measured using the method of Nelson and Sommers (1982). Total Kjeldahl nitrogen was determined by Bremmer and Mulvaney (1982) procedure. Total available phosphorus (TAP) was determined by colorimetric method (Bansal and Kapoor 2000). Total Potassium, Sodium and Calcium was determined by flame photometer (Bansal and Kapoor 2000). The pH and electrical conductivity (EC) were determined by the method of Garg et al. (2006) a double distilled waster suspension of vermiwash/ vermicomposts in ratio of 1:10 (w/v) that had been agitated mechanically for 30 minute and filtered through whatman No.1 filter paper. Statistical Analysis: The data have been expressed as mean SE of 6 replicates. Two way analysis of variance (ANOVA) was applied to determined any significant (P<0.05) difference among the parameters observed in vermicomposts bed (Sokal and Rohlf, 1973). RESULTS AND DISCUSSION The vermicompost was much darker in color than originally and had been processed more or less in to homogenous mixture after 50 to 60 days of earthworms activity. The vermicomposting significantly changed the physio-chemical properties of different feed mixture tested. There were little changed in pH of feeds of all vermibed (Table 1, 2). The pH decreased from alkaline to acidic or neutral (7.60.04- 6.5 0.05). The pH shift toward acidic condition was attributed to mineralization of nitrogen and phosphorus in to nitrates /nitrites and orthro-phosphates; bioconversion of organic materials in to intermediate species of organic acids (Ndegwa et al., 2000). Production of CO 2 and organic acid by microbial decomposition during vermicomposting lowers the pH of substrate (Garg, 2004; Haimi and Hutha, 1986; Elvira et al.,1998; Hartenstien and Hartenstien, 1981).

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Aust. J. Basic & Appl. Sci., 3(4): 3671-3676, 2009 All the treatments show a similar pattern on of change in EC. The initial electrical conductivity in feed mixture were from 0.94 0.04 2.94 0.14 ds/m, while final EC was in ranged from 0.62 0.05 2.300.02ds/m. The EC was reduced in the range of 30 to 48% for different feeds after vermicomposting. The increased in EC might have been due to the loss of weight of organic matter and release of different mineral salts in available forms (W ong et al., 1997; Kaviraj and Sharma, 2003). The organic carbon (TOC) in all vermibeds declined (45 to 50) drastically compared to their initial levels (table 1, 2). Elvira et al, 1998; Kaushik and Garg, 2003 have reported 20-45 % loss of TOC in the form of CO 2 from different industrial sludge during vermicomposting. The vermicomposting process refers to feeding of earthworm on organic matter and microbial degradation. Earthworms modify the substrate condition which consequently promotes the carbon losses from the substrate through microbial respiration in form of CO 2 and even through mineralization of organic matter. Elvira et al.(1998) stated that large fraction of organic matter in the initial substrate was as loss as CO 2 between 20 and 43% as total organic carbon by the end of vermicomposting. Kale et al., 1982; Elvira et al., 1998; Suthar, 2007 reported that body fluid and excreta secreted by earthworm (e.g. mucus, high concentration of organic matter, ammonium and urea) promote microbial growth in vermicomposting. Total Nitrogen content was increased in between 8.0 0.06 and 26.9 0.08 g/kg in final products of different vermibed because of mineralization of organic matter (Table1 and 2). Hand et al.(1988) have been already reported that Eisenia foetida in cow dung slurry increased the nitrate-nitrogen content. Losses of organic carbon might be responsible for nitrogen addition in the form of mucus nitrogenous excretory substances, growth stimulatory hormones and enzymes from the gut of earthworms (Tripathi and Bhardwaj, 2004; Viel et al. 1987). These nitrogen rich substances were not originally present in feed and might have contributed additional nitrogen content. A decrease in pH may also be an important factor in nitrogen retention as this element is lost as volatile ammonia at higher pH values (Hartenstein and Hartenstein, 1981). Atiyeh et al.(2000) reported that by enhancing nitrogen mineralization, earthworms have a great impact on nitrogen transformation in manure, so that nitrogen retained in the nitrate form. The C: N ratio one of the most widely used indices for maturity of organic wastes, decreased with time in all the experiment due to decomposition (table-1,2). Initial C: N ratio was in the range of 42.650.3132.01.2. Final C: N ratios were in the range of 8.8 1.06 27.01.40. According to Senesi (1989) declined in C: N ratio to less than 20 which indicates an advance degree of organic matter stabilization and reflects a satisfactory degree of organic wastes. Suthar, (2008) reported that the C: N ratio of substrate material refleets the organic waste mineralization and stabilization during the process of decomposition. The loss of carbon as CO 2 through microbial respiration and added of nitrogenous excretory material between the C: N ratio of the substrate. C: N ratio is one of the most widely used indicators of vermicompost maturation, decreases sharply during vermic process (Kale, 1998, Gupta and Garg, 2007, Suther, 2008). Total phosphorus was greater in final vermicomposts than initial vermibed mixture (Table 1,2). The maximum TP was observed in combination of horse dung with wheat bran 13.20.01 g/ Kg followed by horse dung with rice bran. It is because of the physical breakdown of the earthworms (Mansell et al., 1981). The increase of 25 % in TP of paper waste sludge, after worm activity is performed partly by earthworm gut phosphates and further release of TP and further release of TP might be attributed to the P-solubility microorganisms present in warm casts ( Satchell and Martin, 1984).According to Lee (1992) the passage of organic residue through the gut of earthworm to the plant the released of phosphorus in available form is performed partly by earthworm gut phosphatases and further released of phosphorus might be attribute to phosphorus solubilizing micro organism present in worm cast. Total potassium concentration was slightly increased in vermicomposts/vermiwash of all the combination of wastes than initial feed mixture (Table1, 2). Delgado et al. (1995) have reported a higher content of TK in the new sewage sludge vermicompost. Benitez et al., (1999) studied that the leacheates collected during vermicomposting process had higher K concentrations. Kaviraj and Sharma (2003) observed that level of TK was increased 10% by Eisenia foetida and 5% by L. maturitii during vermicomposting. Shutar (2007) suggested that earthworm processed waste material contains high concentration of exchangeable K, due to enhanced microbial activity during the vermicomposting process, which consequently enhanced the rate of mineralization. Total Calcium concentration was higher in final products than initial feed mixture (table1, 2). Elvira et al. (1996) have reported no significant increased in T Ca for vermicomposting of paper mill sludge. It is suggested that gut process associated with calcium metabolism are primarily responsible for enhanced content of inorganic calcium content worm cast. However, the similar pattern of calcium enhancement is well documented in available literature (Heartenstin and Heartenstin, 1981; Garg et al. 2006).

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Table 1: N utrient content and different physio-chem ical param eter in verm iwash obtained from initial feed m ixture and final Verm icom posts of different com binations of anim al and agro / bitchen wastes. Com bination Initial feed m ixture ----------------------------------------------------------------------------------------------------------------------------------------TOC (g/kg) TKN (g/kg) C:N ratio TK (g/kg) TP (g/kg) pH EC ds/m TCa (g/kg) Cow D ung 480.354.21 6.00.02 86.22.40 4.70.07 3.70.03 8.00.02 2.150.03 1.30.2 D ung+Gram Bran 598.022.21 13.20.68 45.20.26 7.00.02 4.80.08 3.00.03 2.610.06 2.00.1 D ung+ Straw 635.044.32 8.60.08 73.80.04 6.40.08 3.70.03 8.80.08 2.890.08 2.10.3 D ung + W heat Bran 681.003.04 12.81.00 53.21.03 6.90.10 7.70.04 8.40.03 2.630.07 1.50.2 D ung + Rice Bran 568.102.32 11.00.32 51.60.23 5.80.12 6.90.08 8.70.04 2.740.12 1.30.4 D ung + Vegetable W astes 569.122.05 9.80.62 58.60.86 6.80.08 4.20.05 8.00.04 2.460.02 1.60.4 D ung + Barley Bran 482.142.03 10.40.04 46.31.20 5.90.12 4.90.01 8.30.02 2.800.06 1.20.1 Buffalo D ung 514.462.20 6.40.02 92.01.60 6.90.16 4.90.0 8.50.08 2.590.06 1.30.2 D ung + Gram Bran 647.122.42 13.70.37 47.10.38 7.60.12 6.90.04 8.60.02 2.640.12 2.20.2 D ung + Straw 684.023.14 8.80.06 77.11.08 7.30.16 4.90.02 8.50.14 2.940.13 1.60.4 D ung + W heat Bran 734.002.62 13.40.10 54.81.62 8.60.17 9.00.03 8.20.05 2.470.08 1.30.4 D ung + Rice Bran 615.012.06 11.30.31 54.20.84 8.90.12 8.20.02 8.40.06 2.760.13 1.80.6 D ung + Vegetable W astes 574.301.37 10.00.10 57.40.82 8.70.18 5.70.06 8.50.04 2.780.12 1.50.8 D ung + Barley Bran 509.062.32 11.80.04 43.11.36 8.00.13 6.30.07 8.00.04 2.960.18 1.60.3 Goat D ung 432.903.12 4.50.20 93.00.13 6.20.14 4.00.04 8.50.03 2.540.16 1.60.5 D ung + Gram Bran 556.003.17 12.80.25 43.41.20 7.00.14 5.60.09 8.50.06 2.480.12 1.50.4 D ung + Straw 596.120.62 7.20.06 78.90.09 6.80.18 4.50.05 8.50.20 3.000.12 2.20.1 D ung + W heat Bran 601.003.05 11.00.31 54.600.5 8.00.12 8.50.06 8.30.04 2.800.14 1.30.3 D ung + Rice Bran 530.001.42 10.20.32 48.500.7 7.90.20 7.60.02 8.50.04 2.800.08 1.20.5 D ung + Vegetable W astes 535.712.58 8.70.10 61.601.7 8.30.16 5.20.04 9.00.02 2.850.12 1.30.2 D ung + Barley Bran 468.702.87 9.00.13 52.001.2 7.80.15 5.40.06 9.00.08 2.740.16 1.70.4 Com bination Final Verm icom posts ----------------------------------------------------------------------------------------------------------------------------------------TOC (g/kg) TKN (g/kg) C:N ratio T K (g/kg) TP (g/kg) pH EC ds/m TCa (g/kg) Cow D ung 199.000.84 15.20.05 13.11.71 4.80.07 7.00.05 6.80.31 1.300.02 1.90.3 D ung+Gram Bran 263.601.08 24.10.04 10.00.42 7.40.08 7.30.40 6.90.06 1.080.08 2.30.4 D ung+ Straw 218.405.46 18.00.25 12.10.62 6.80.02 5.50.05 7.20.03 1.200.06 2.20.3 D ung + W heat Bran 303.204.21 25.00.22 12.20.25 7.60.05 9.80.06 7.30.02 1.100.04 2.40.1 D ung + Rice Bran 252.405.02 24.10.45 10.40.48 6.30.06 8.70.05 7.60.05 1.080.02 2.60.2 D ung + Vegetable W astes 245.702.25 21.90.08 11.20.01 7.40.10 6.60.05 7.30.02 1.080.06 2.30.3 D ung + Barley Bran 216.402.06 24.60.08 8.81.06 6.40.02 7.40.05 7.20.01 1.160.04 2.10.3 Buffalo D ung 260.201.04 10.70.02 24.31.03 7.20.08 6.00.02 7.00.06 1.420.04 2.80.3 D ung+Gram Bran 318.002.13 23.92.26 13.31.06 8.10.13 7.50.06 7.00.06 1.040.08 4.20.4 D ung+ Straw 278.402.32 17.50.12 15.90.04 8.00.15 5.60.04 6.70.04 1.310.10 4.40.3 D ung + W heat Bran 390.101.52 22.30.40 17.50.82 8.30.04 10.40.10 7.40.08 1.190.11 4.00.3 D ung + Rice Bran 340.282.48 23.60.42 12.90.08 9.00.12 9.00.08 6.50.04 1.150.02 5.80.1 D ung + Vegetable W aste 310.011.23 21.50.14 14.40.73 8.50.21 6.80.05 7.30.03 1.170.18 3.60.6 D ung + Barley Bran 294.252.45 21.00.06 14.11.40 8.70.15 7.90.08 7.40.06 1.260.17 3.50.4 Goat D ung 228.011.62 09.00.04 25.31.40 6.80.10 5.20.08 6.70.04 1.730.12 2.50.3 D ung + Gram Bran 298.000.08 24.40.14 12.20.81 8.70.14 7.00.04 7.20.01 1.200.08 3.20.4 D ung + Straw 254.061.37 18.40.04 13.81.20 7.20.05 4.80.08 7.00.02 1.460.02 3.60.2 D ung + W heat Bran 362.021.15 26.90.60 13.50.20 8.60.17 8.30.06 7.00.02 1.280.15 3.00.1 D ung + Rice Bran 298.002.80 24.60.12 12.11.16 8.10.16 8.20.04 7.30.03 1.230.05 3.40.4 D ung + Vegetable W astes 272.013.02 22.00.12 12.41.60 8.70.12 6.20.04 6.50.10 1.240.07 3.60.3 D ung + Barley Bran 254.002.53 25.10.14 10.10.81 8.30.13 6.20.10 7.10.04 1.390.17 3.00.2 Each value is the m ean SE of six replicates. Significant variance (P<0.05) two way analysis of variance (AN O VA) was applied in between the different param agnet of verm iwash of initial feed m ixture and Final Verm icom posts. Table 2: N utrient content and different physio-chem ical param eter in verm iwash obtained from initial feed m ixture and final Verm icom posts of different com binations of anim al and agro / bitchen wastes.s Com bination Initial feed m ixture ----------------------------------------------------------------------------------------------------------------------------------------TO C (g/kg) TKN (g/kg) C:N ratio TK (g/kg) TP (g/kg) pH EC ds/m TCa (g/kg) Sheep D ung 321.011.80 4.10.08 85.810.8 6.90.12 3.00.05 9.00.06 0.940.04 1.30.3 D ung + Gram Bran 542.601.34 12.30.13 42.650.3 7.30.14 5.40.06 8.60.13 2.160.06 1.50.5 D ung + Straw 612.001.43 7.40.04 83.01.40 7.40.12 4.20.03 8.30.04 2.280.02 1.30.4 D ung + W heat Bran 589.513.36 10.20.04 57.81.20 7.80.12 8.00.06 8.30.04 2.180.06 1.20.1 D ung + Vegetable W astes 498.601.62 8.40.06 58.40.60 7.70.06 4.70.02 9.00.07 2.140.10 1.50.2 D ung+ Rice Bran 440.012.52 10.50.08 42.90.80 7.20.08 7.80.06 8.30.00 2.210.08 1.20.3 D ung + Barley Bran 438.001.42 8.20.01 54.81.20 7.20.08 5.90.06 8.00.04 2.240.18 1.60.2 H orse D ung 470.021.34 4.00.08 132.01.20 7.90.06 6.80.03 8.10.06 2.100.04 1.30.5 D ung + Gram Bran 627.011.42 12.50.62 50.20.20 7.70.06 8.70.05 7.90.14 2.690.12 1.50.2 D ung + Straw 650.213.10 7.20.12 89.30.12 7.90.08 5.50.07 8.50.14 2.940.12 1.20.2 D ung + W heat Bran 715.013.13 10.60.06 67.51.40 7.90.09 11.10.1 8.00.06 2.740.15 1.40.5 D ung + Rice Bran 584.043.14 10.30.15 56.71.80 7.20.08 10.80.1 8.50.08 2.680.12 1.20.4 D ung + Vegetable W astes 560.211.11 8.100.21 69.130.6 7.90.04 7.80.06 8.20.10 2.730.14 1.50.5 D ung + Barley Bran 492.101.45 8.200.12 60.00.80 8.00.05 8.20.06 8.40.04 2.880.12 1.20.3

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Table 2: Continue Com bination

Final Verm icom posts ----------------------------------------------------------------------------------------------------------------------------------------TO C (g/kg) TKN (g/kg) C:N ratio TK (g/kg) T P (g/kg) pH EC ds/m TCa (g/kg) Sheep D ung 216.012.62 08.00.06 25.70.60 7.40.13 6.20.08 7.30.08 0.780.02 3.10.3 D ung + Gram Bran 234.212.62 23.20.20 11.01.00 7.90.17 7.20.04 7.20.06 0.690.02 5.20.2 D ung + Straw 208.002.34 17.10.25 12.21.05 7.80.14 5.30.04 7.00.10 0.700.04 5.20.4 D ung + W heat Bran 335.002.47 23.40.14 14.30.80 8.10.12 9.30.09 6.50.10 0.620.05 4.30.8 D ung + Vegetable W astes 212.402.52 21.60.32 09.80.09 8.20.16 6.30.04 7.40.04 0.640.02 3.20.7 D ung+ Rice Bran 280.071.45 21.60.32 9.80.09 8.20.16 6.30.04 7.40.04 0.640.08 3.40.6 D ung + Barley Bran 230.112.43 21.20.20 10.91.30 8.00.08 6.80.04 6.90.02 0.680.02 3.20.4 H orse D ung 215.152.02 8.00.14 27.01.40 8.00.06 9.90.05 7.00.02 1.270.08 3.80.6 D ung + Gram Bran 252.011.82 21.00.25 12.00.60 8.00.02 10.30.06 7.00.06 1.060.05 5.20.8 D ung + Straw 200.711.02 16.70.42 12.00.22 8.40.10 7.90.06 7.30.17 1.270.08 5.60.2 D ung + W heat Bran 348.302.17 22.00.42 15.81.50 8.20.12 13.20.02 6.90.04 1.900.05 5.00.3 D ung+ Rice Bran 260.411.62 22.00.82 11.80.61 7.90.06 12.60.07 6.50.10 1.200.03 5.00.4 D ung + Vegetable W astes 239.011.41 20.80.14 11.50.3 8.20.06 9.00.08 7.40.02 1.150.05 4.40.3 D ung + Barley Bran 228.102.32 20.50.06 11.10.2 8.60.09 10.80.10 7.20.08 1.240.04 3.80.6 Each value is the m ean SE of six replicates. Significant variance (P<0.05) two way analysis of variance (AN O VA) was applied in between the different param agnet of verm iwash of initial feed m ixture and Final Verm icom posts.

Conclusion: It is evident from results the significant increase of TKN, TP, TK and TCa and significant decrease in TOC, C:N ratio, EC and pH in final vermicompost of different animal/ agro and kitchen wastes by the use of earthworm Esinia foetida is due to the vermic activity in laboratory conditions. It will suggested that the final product of which combination having potential amount of nutrients and that recognized combinations of wastes are suitable for particular deficient soil as biofertilizer for organic farming and also help in obtaining chemical composition of vermicomposts. Thus vermicomposting as an best technology for the recycling of wastes and production of biofertilizers from different wastes by use earthworms Eisenia foetida . ACKNOW LEDGM ENT Authors are thankful to U.G.C., New Delhi project F.No.33-351/2007 (SR) for financial assistance. REFERENCES Atiyeh, R.M., J. Dominguez, S. Subler and C.A. Edwards, 2000. Change in the biochemical properties of cow manure during processing by earthworm Eisenia andrei (bouch) and the effect of seeding growth. Pedobiologia, 44: 709-724. Aira, M., F. Monoroy, J. Dominguez, and S. Mato, 2002. How earthworm density affects microbial biomass and activity in pig manure. Eur. J. Soil Biol., 38: 7-10. Bremmer, J.M. and R.G. Mulvaney, 1982. Nitrogen Total in: A.L. page R.H. Millar and D.R. Keeney, (eds.), Method of Soil Analysis, American Society of agronomy, Madison, pp: 575-624. Bhatnagar, R.K. and R.K. Paitta, 1996. Earthworm: Vermiculture and Vermicomposting; Kalyani Publishers, Ludhiana. Bansal, S. and K.K. Kapoor, 2000. Vermicomposting of crop residues and cattle dung with. Eisenia foetida. Biores. Technol., 73: 95-98. Benitez, E., R. Nogales, C. Elvira, G. Masciandaro and B. Ceccanti, 1999. Enzyme activity as indicators of the stabilization of sewage sludge composting with Eisenia foetida. Biores Technol., 67: 297-303. Bentize, E., R. Nogales, G. Masciandro and B. Ceccanthi, 2000. Isolation by isoelectric focusing of humic urease complex from earthworm Eisenia foetida processed sewedge sludge. Biol. Fert. Soil., 31: 489-493. Benitz, E., H. Sianz and R. Nogales, 2005. Hydrolytic enzyme activities of extracted humic substances during the vermicomposting lignocelulosic olives waste. Biores. Technol., 96: 785-790. Chaudhari, P.S. and G. Bhattacharjee, 2002. Capacity of various experimental diets to support biomass and reproduction of Perionyx excavatus Biores. Technol., 82: 147-150. Delgado, M., M. Bigeriego, I. W alter and R. Calbo, 1995. Use of California red worm sewage sludge transformation. Turrialba., 45: 33-41. Elvira, C., M. Goicoechea, L. Sampdro, S. Mato and R. Nogalas, 1996. Bioconversion of solid paper pulp mill sludge by Earthworm, Biores. Technol., 75: 173-177. Elvira, C.A., L. Sampedro, E. Benitez and R. Nogales, 1998. Vermicomposting of sludges from paper mill and dairy industries with Eisenia Andrei: Apiolet scale study. Biores. Technol., 63: 205-211. Edwards, C.A., J. Dominguez and E.F. Neuhauser, 1998. Growth and reproduction of Perionyx excavatus (Perrier) (Megascolicideae) as factor inorganic waste management. Biol. Fert. Soils., 27: 155-161. 3675

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