International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202Vol. 3(2), 70-73, February (2014) Int. Res. J. Biological Sci. International Science Congress Association 70 Bioconversion of Municipal Solid Waste by the Earthworm Eudrilus eugeniae (Kinberg)Sriramulu Ananthakrishnasamy and Govindarajan Gunasekaran Department of Zoology, Annamalai University, Annamalai Nagar – 608 002, Tamil Nadu, INDIAAvailable online at: www.isca.in, www.isca.me Received 31st October 2013, revised 1st December 2013, accepted 6th January 2014Abstract Rapid urbanization, industrialization and agricultural practices have led to dumping of organic solid wastes caused a serious threat to the environment. In the present investigation an attempt has been made to convert the municipal solid waste (MSW) into manure by the earthworm Eudrilus eugeniae under laboratory conditions. The experimental media were prepared on dry weight basis by mixing the municipal solid waste and bedding material (BM); E– 20% BM + 80% MSW, – 40% BM + 60% MSW, E – 60% BM + 40% MSW, E – 80% BM + 20% MSW, controls (BM alone – C) were also maintained. The growth rate (biomass) and reproduction (number of cocoons and number hatchlings) of E. eugeniae was increased in all experimental media BM + MSW mixtures (E – E) and control (C). The result showed that the unutilized and enormously available MSW can be vermicomposted into valuable organic manure that can be utilized for sustainable agriculture. Keywords: Bedding material, municipal solid waste, Eudrilus eugeniae, Cocoons, Hatchlings. Introduction A rapid development in industries and growth in population in India has released to generate thousand tons of municipal solid waste (MSW) every day. Solid waste management (SWM) is a major environmental problem of our countries. India produces around 3000 million tonnes of organic wastes annually. The processing of MSW by vermicomposting is an eco-friendly approach result in the production of organic fertilizr. The growth (biomass) and reproduction (cocoons and hatchlings) of earthworm in different culture media such as mixture of animal and vegetable wastes; kitchen waste; animal wastes; water hyachinth; different organic wastes; baggase (sugar industrial waste) were studied. The survey of literature indicated that reports on growth and reproduction are available more on organic wastes but information about growth and reproduction of earthworm on municipal solid waste (MSW) are scanty. Hence it was aimed to study the growth and reproduction of earthworm and which will help us to have an idea about best combination of bedding material (BM) with MSW. Material and Methods The breeding stocks of Eudrilus eugeniae weremaintained in the Department of Zoology, Annamalai University. Cow dung was used as substrate to maintain the adult worms; moisture content of 60 – 70% was continuously maintained by sprinkling water on the stock culture of the cement tank. This stock culture in the cement tank were covered with gunny bag and maintained at room temperature (27± 2°) inside the animal house. MSW was collected from Sethiathope town Panchayat, Cuddalore District, Tamil Nadu. After removing the plastics, polythene, metal scraps and glass pieces MSW was dried and brought by using jute bags to the laboratory. Urine and straw free cow dung was collected from dairy yard at the Faculty of Agriculture, Annamalai University. It was sun dried, powdered and stored in jute bags. The pressmud was collected from M.R.K Cooperative Sugar Mill, Sethiathope. The collected pressmud was cured for a month to remove the odour. Then it was used for the preparation of Bedding Material (BM). The cow dung and one month old cured pressmud was used for the preparation of bedding material and they were equally mixed on dry weight basis and kept as such for 15 days and used for the preparation of substrate for vermiculture. Combinations of bedding material and municipal solid waste in four proportions were prepared in the following orders: C – BM (Bedding material) alone (Control) 500 gram CD + 500 gram PM + 200 gram soil; 1. E1 - 20% + 80% (BM + MSW) 200g BM + 800g MSW + 200g soil; 2. E2 40% + 60% (BM + MSW) 400g BM + 600g MSW + 200g soil; 3. E3 - 60% + 40% (BM + MSW) 600g BM +400g MSW+ 200g soil; 4. E4 - 80% + 20% (BM + MSW) 800g BM + 200g MSW+ 200g soil mixed W/W. After the preparation of substrates in the above different proportions, water was sprinkled and kept as such for thermophilic composting for 15 days. After the completion of thermophilic composting fifteen grams of sexually mature, clitellate Eudrilus eugeniae (approx. 38 days old) were introduced in plastic troughs separately; containing 1 Kg substrate + 200 g of soil. Bedding material alone was used as control, separately for E. eugeniae as C. Six replications in each experimental treatment have been maintained for 60 days. Earthworm’s growth (biomass) and reproduction (Cocoon and International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 3(2), 70-73, February (2014) Int. Res. J. Biological Sci. International Science Congress Association 71 hatchling number) were recorded in different time intervals like 15, 30, 45 and 60 days. Every time cocoons and hatchlings were counted by hand sorting and the biomass of worms were weighed using electronic balance and recorded. For earthworm’s mean biomass and reproduction (cocoon and hatchlings) standard deviation (SD), percentage increase or decreases (final biomass) over initial values were calculated. Further, the data were analyzed statistically (significance of difference of 0.05 levels) by using two - way analysis of variance, (ANOVA). Results and Discussion Growth (biomass) and reproduction (cocoon and hatchling) of E. eugeniae cultured on different feed substrates are given in table – 1 to 3. Table - 1 shows the obtained values of growth (biomass) of E. eugeniae in different treatment media over the study period. Initially all the experimental media i.e. C, E, E, and E more or less 15 g of worms were weighed and inoculated. Thereafter the growth rate was gradually increased on 15th, 30th, 45th and 60th day. The maximum growth rate was recorded in E3 (18.1 ± 1.19) followed by C(17.2 ± 1.12), E(16.9 ± 1.12), E (16.2 ± 1.21) and E (15.7 ± 1.16) respectively on 15th day. The growth rate was steadily increased upto 60thday. The highest growth rate was obtained on 60th and it was recorded in E (24.7 ± 1.15), followed by C (22.4 ± 0.94), E(20.9 ± 1.18), E2 (19.9 ± 1.14) and E (18.8 ± 0.99) respectively. Among the treatments the highest growth was recorded in Eand least growth was recorded in E. The present study clearly indicated that the growths of E. eugeniae were found in the following order: E� C � E � E � E. The production of cocoons by E. eugeniae in different feed substrates are presented in table - 2. The adult clitellate worms started to produce of cocoons and they were found on 15th day in all treatments. On 15th day the maximum number of cocoons were recorded in E3 (10.4 ± 0.90) and followed by C (9.1 ± 1.15), E4 (8.2 ± 1.23), E (7.6 ± 0.90) and E (7.2 ± 0.95) respectively. The cocoons production was gradually increased after 15th day till 60th day. (i.e. end of the experimental period). The highest production of cocoons were recorded on 60th day in (20.5 ± 1.16) followed by C (18.7 ± 1.19), E4 (16.3 ± 1.14), 2 (14.4 ± 1.17) and E1 (13.6 ± 1.21). More number of cocoons were observed in E (60.4) and least was observed in E (39.6). The production of cocoons were in the order: E � C� E � E � . The production of hatchlings of E. eugeniae are presented in table - 3. Hatchlings were observed on 30th day in all treatments. From 30th day to 60th day the hatchling numbers were increased. More number of hatchlings were recorded in E (19.9 ± 1.10), followed by C (17.7 ± 1.12), E (15.8 ± 0.99), E (14.5 ±1.11) and E1 (13.0 ± 1.18) on 30th day. Thereafter 45th day also increased gradually. On 60th day the numbers of hatchlings obtained were 32.3 ± 1.17 in E, 23.8 ± 1.36 in C, 21.4 ± 1.18 in , 19.2 ± 1.25 in E and 17.9 ± 1.21 in E respectively. Among all the treatments the highest production of hatchlings were found in E3 (76.6). Table-1 Growth of E. Eugeniae during the vermicomposting of MSW (p0.05) Substrate ProportionsE. eugeniae Vermicomposting Days 0 (Initial) 15 30 45 60 C 15.0±1.17 17.2±1.12 19.7±1.11 21.4±1.18 22.4±0.94 (49.33) 15.2±1.27 15.7±1.16 17.1±0.95 18.0±1.19 18.8±0.99 (23.68) 15.1±1.19 16.2±1.21 17.9±0.98 19.2±1.14 19.9±1.14 (31.79) 14.9±0.95 18.1±1.19 21.4±0.99 23.5±1.19 24.7±1.15 (65.77) 15.3±1.23 16.9±1.12 19.2±1.19 20.3±1.12 20.9±1.18 (36.60) ANOVA Analysis of variance Sum of square Df Mean of square F-value P-value Rows 37.748 4 9.437 9.561297 0.00038 Columns 133.32 4 33.33 33.769 1.29E-07 Error 15.792 16 0.987 C – Control (BM alone), E1 – (20% BM + 80% MSW), E2 – (40% BM + 60% MSW), E3 – (60% BM + 40% MSW), E– (80% BM + 20% MSW), Initial (0) – Worm unworked substrates, Mean SD of six observations, (+/–) – Percent change of increase or decrease over the initial. International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 3(2), 70-73, February (2014) Int. Res. J. Biological Sci. International Science Congress Association 72 Table-2 Cocoons laid by E. eugeniae during the vermicomposting of MSW (p0.05) Substrate ProportionsE. eugeniae Vermicomposting Days 0 Initial 15 30 45 60 Total No. of Cocoons C - 9.1±1.15 10.3±0.98 13.6±0.90 18.7±1.19 51.7 E 1 - 7.2±0.95 8.7±1.14 10.1±1.17 13.6±1.21 39.6 E 2 - 7.6±0.90 8.7±1.12 10.8±1.12 14.7±1.17 41.8 E 3 - 10.4±0.90 12.6±1.19 16.9±1.16 20.5±1.16 60.4 E 4 - 8.2±1.23 9.0±1.14 11.2±1.20 16.3±1.14 44.7 ANOVA Analysis of variance Sum of square Df Mean of square F-value P-value Rows 71.673 4 17.91825 23.57923 1.31E-05 Columns 198.626 3 66.20867 87.12622 2.06E-08 Error 9.119 12 0.759917 C – Control (BM alone), E– (20% BM + 80% MSW), E2 – (40% BM + 60% MSW), E3 – (60% BM + 40%, MSW), E– (80% BM + 20% MSW), Initial (0) – Worm unworked substrates, Mean SD of six observations, (+/–) – Percent change of increase or decrease over the initial.Table-3 Hatchlings of E. eugeniae during the vermicomposting of MSW (p0.05) Substrate ProportionsE. eugeniae Vermicomposting Days 15 30 45 60 Total No. of Hatchlings C 0 17.7±1.12 18.6±0.97 23.8±1.36 60.1 E 1 0 13.0±1.18 15.2±1.21 17.9±1.21 46.1 E 2 0 14.5±1.11 15.4±1.25 19.2±1.25 49.1 E 3 0 19.9±1.10 24.4±1.24 32.3±1.17 76.6 E 4 0 15.8±0.99 17.3±1.20 21.4±1.18 54.5 ANOVA Analysis of variance Sum of square Df Mean of square F-value P-value Rows 145.212 4 36.303 6.190473 0.006099 Columns 1486.908 3 495.636 84.51702 2.45E-08 Error 70.372 12 5.864333 C – Control (BM alone), E1 – (20% BM + 80% MSW), E2 – (40% BM + 60% MSW), E3 – (60% BM + 40% MSW), E– (80% BM + 20% MSW), Initial (0) – Worm unworked substrates, Mean SD of six observations. (+/–) – Percent change of increase or decrease over the initial. In the present investigation it was clearly observed that the highest growth and reproduction was observed in E. eugeniae and itshowed the maximum growth rate in E3. The growth of earthworm are depend on the quality of the available food, adequate temperature and moisture found. The attainment of sexual maturity in E. eugeniae was reported between 35 - 49 days and between 35 - 45 days cultured on cattle manure10. The production of cocoons in E. eugeniae at the age of 46 days at 25C in fresh urine free cattle manure11. A linear relationship with size of the worm and cocoon size were noted12. Growth and reproduction in earthworms require OC, N and P which is obtained from litter, grit and microbes13. The N - content played a major role in the biomass gain and hatchlings success but not have affected the cocoon production14. The higher feeding rate of E. eugeniae, they had higher reproductive potential of 17 cocoons and 550 juveniles per 100g of compost in 41 days exceeded that for L. mauritii of 7 cocoons and 400 juveniles in 56 days reported15. Comparing our results with others in literature, the E. eugeniae was able to produce more number of cocoons and hatchlings in E which are considered to be the best combination for the earthworm respectively. The reason for higher growth and reproductive rate shown by the earthworm i.e. in E by E. eugeniae in the present study might by due to i. Rich in nitrogen and phosphorus in the experiment (E), ii. Highest microbial population, iii. Increased enzymatic activity which enables the earthworm for their suitability. The above results clearly suggest that the incorporation of bedding material (Pressmud and Cowdung) in International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 3(2), 70-73, February (2014) Int. Res. J. Biological Sci. International Science Congress Association 73 an appropriate ratio not only increases the selective palatability but also enhances the growth and reproduction of earthworms. The mixing of BM increased the suitability of MSW as feed substrate for microbes and earthworms16. Conclusion The over population and industries in developed and developing countries have led to the production of large volumes of municipal solid waste (MSW). Earthworms have successful development in cow dung (CD) and pressmud (PM), hence in the present study the bedding material (BM) was prepared from CD and PM and mixed with MSW in different proportions to facilitate the earthworms to decompose the MSW. 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