MINOR RESEARCH PROJECT

Project Title “A novel Vermitechnology approach to improve the soil fertility by the Production of Vermibricks - Farmers user-friendly.”

UGC Reference No. F MRP-6648/16 (SERO/UGC) Dated 30th June 2017 Period of report: from 30.06.17 to 31.07.19

Final Report of the work done on the Minor Research Project

1. Introduction

Soil provides food, fodder and fiber and 99 % of our food comes from land. Only 22 % of our global land area (13.9 x 10 9 ha) is suitable for agriculture and only 3 % is suitable for high level of productivity. The low level of yield in the developing countries is due to poor management of soil, water and crop. There is an essential link between soil, plant, and man. An understanding of this vital ecological balance is the pre requisite not only for sustaining agriculture practices but also for a healthy life of man.

The old traditional agriculture practice, in the tropical region was based on some kind of rotational system and was entirely dependent on soil organic matter and manure, particularly farmyard manure. The modern agriculture practice, intensive and continuous due to increased human population and pressure on land, is based on development of improved varieties of crops, chemical fertilizers and pesticides. But the modern agricultural practices necessitate the use of a wide range of chemicals, which may adversely affect non-target organisms. These often exposed to a wide range of anthropogenic compounds released into the terrestrial environment; as a consequence, they suffer from the toxicity of these compounds. The abundance or activity of has been considered as an indicator of the biological health of soil.

In such a context it is essential that agriculture practices give priority to agro-forestry or herbaceous legumes based cropping system based on local conditions and regulate soil fertility by biological processes. This could be done by increased efficiency of the use of natural resources, which is otherwise known as organic farming. Vermicomposting is one of the practices of organic farming.

Objective of the project:

✓ To facilitate the farmers to use the vermicompost in a user friendly and potential way. ✓ To facilitate the entrepreneurs who do organic manure business and to provide them a holistic view of business. ✓ To reduce the manpower in the farms. ✓ To retain the moisture and microbial inoculums. ✓ To reduce pollution and provide a valuable substitute for chemical fertilizers in the soil.

2. Review of Literature

The success of sustainable agriculture is very much dependent upon the availability of cheap and good quality organic manure. Among the source of available organic manures vermicompost is a potential source due to the presence of readily available plant nutrients, growth enhancing substances and number of beneficial microorganisms. Microorganisms and are important biological organisms helping nature to maintain nutrients flow and their role in restoration of contaminated soil is not fully known. Vermicomposting and its use offers several potential benefits including improved manure handling, enhanced soil tilts and fertility, and reduced environmental risk. The farmers have realized the importance of vermicomposting and particularly its eco friendly nature and in making sustained availability of nutrients to soil due to vermicompost application.

Vermicomposting is the process of production of organic manure by the decomposition of organic wastes facilitated through earthworms interacting with microorganisms. Since earthworms are saprophagous, detritivorous, geophagous and microbivorous, they consume a wide variety of biodegradable and convert them into good organic manure. Wide range of agricultural residues, such as straw, husk, leaves, stalks, weeds etc can be converted into vermicompost. Other potential feedstock for vermicompost production are livestock wastes, poultry litter, dairy wastes, food processing wastes, organic fraction of MSW, bagasse, digestate from biogas plants etc. Almost all types of biologically degradable and decomposable organic wastes are used in vermiculture and vermicomposting. Commonly used composting materials are, animal dung like cattle dung, sheep dung, horse dung, agricultural wastes like stem, leaves, husk of grains, peels, vegetable waste, orchid leaf litters, processed food wastes, sugarcane trash and baggase, forestry wastes like wood savings peels, saw dust and pulp leaf litters waste paper and cotton cloth, city refuge or garbage kitchen wastes, biogas slurry and industrial wastes. Earthworms consume organic wastes and reduce the volume by 40–60 percent. Each earthworm weighs about 0.5 to 0.6 gram, eats waste equivalent to its body weight and produces cast equivalent to about 50 percent of the waste it consumes in a day. The moisture content of castings ranges between 32 and 66 percent and the pH is around 7. The level of nutrients in compost depends upon the source of the raw material and the species of earthworm.

Eudrilus eugeniae, has been effectively utilized for vermicomposting of organic waste materials like cassava peel (Mba, 1983), poultry manure, press mud (Ramalingam, 1997) leaf litters (Daneil and Karmegam, 1999) tea leaves waste, vegetable waste and fruit waste (Jayashree et al., 2008).

Worm casts have been shown to exhibit more enzymatic and microbial activities and NPK enrichment (Parthasarathi and Ranganathan, 1998) and also demonstrated an increase in cellulolytic, hemicellulolytic, amylolytic and nitrifying bacteria in worm casts compared to the surrounding soil. The worm castings contain higher percentage of both macro and micronutrients than the garden compost. Apart from other nutrients, a fine worm cast is rich in NPK which are in readily available form and are released within a month of application. Vermicompost enhances plant growth, suppresses disease in plants, increases porosity and microbial activity in soil, and improves water retention and aeration. Vermicompost also benefits the environment by reducing the need for chemical fertilizers and decreasing the amount of waste going to landfills. Vermicompost is essentially a colloid and can hold water up to 9 times its own weight. This can be a great deal of help when the climate has been dry for a long time.The evaporation of water is slow because the water is held up at an organic grade, and therefore, it can be sufficiently utilized by the plants. The above mentioned review give the clear area to be studied and focus the need of the hour.

3. Materials and methods 3.1.Vermicomposting - Selection of suitable species

Selection of suitable species for composting and other possible commercial utilization, viz., eugeniae. Systematic position of : Kingberg Phylum: Annelida Subphylum: Class: Order: Family: Genus: Eudrilus Species: eugenia 3.2 Distribution of Eudrilidae

The distribution of the Eudrilidae is restricted to the Ethiopian region. Eudrilus eugeniae, a widely distributed peregrine species, was once recorded from India near Pune, in the Western Ghats. Originally distributed in equatorial West Africa, presently found distributed in most parts of the world having got introduced for various usages in vermitechnology. It is common in parts of America and Europe and is popularly known as ‘Night Crawler’. In India, common in many vermiculture establishments, particularly in Southern India is recommended species for vermiculture. Since 1930s, species is reportedly distributed in Trivancore, Pune and North Konkan. 3.3 Earthworm culture and Vermicomposting

E.eugeniae were procured from the Sree Sai Agro product, Coimbatore, brought to the Herbal garden and mass cultured in a culture tank containing urine free cow dung. Cow dung was collected from nearby cattle shed, sun dried and maintained. Worms were acclimatized in cow dung. (i) Collection of organic wastes

The waste materials used for composting were cow dung (CD) and leaf litters. The fallen leaves were collected from the college campus and stored in a preparatory tank. All the waste material was mixed with cow dung slurry, kept for15 days for pre decomposition in large tanks. Appropriate moisture level was maintained.

(ii) Vermibed preparation

The pre-decomposed waste materials from the preparation tank were taken for the vermibed preparation. The vermibeds were prepared with cow dung and pre-decomposed. Water was sprinkled over the vermibeds to hold the moisture content of 60 % to 70 % and kept for 24 h. Ten kg of healthy, matured E.eugeniae of 10 to 12 cm length and 0.4 to 0.8 g weight were introduced into the tank. Appropriate controls were maintained. The tanks were covered with perforated sacks for ventilation and for the prevention of predators.

The experimental set up was as follows: Substrates Sample (S1) - Cow Dung + leaf litters (S2) - Cow Dung + leaf litters + E.eugeniae The experiments were carried out at ambient room temperature (28 ± 2° C). The moisture level was maintained throughout the study at 60 % to 70 %. The experimental set up was kept undisturbed for 90 days. The mixture was turned periodically without disturbing the worms, so that the temperature was maintained, and to have uniform maturation of the compost. Observations were made. The experiment was terminated at the end of 90th day. Physico- chemical parameters of vermicompost were analysed on initial and final day. (iii) Particle size, smell and color The pore size, texture, smell and color of the mature compost made by different treatments was observed. (iv) Determination of Moisture percentage of compost The china dish was weight with 10g of compost sample. The china dish was placed in an oven at 120° C for minimum 4 hours. The china dish was taken out after 4 hours of heating and cooled in desiccators. The sample was weight again after heating. Moisture content was calculated by dividing the reduction in weight by initial weight. (iv) Determination of pH The compost solution was made by adding distilled water in 1:10 and for dissolving the maximum salts, it was placed for 2 hours. The pH meter electrode was dip in the compost solution. Reading was noted on pH meter when it was stabilized. The electrode was washed with distilled water and dried with tissue paper (v) Determination of electrical conductivity of compost The solution of compost was made by adding distilled water in compost sample in 1:5. The conductance cell was washed with distilled water. The cell was dipped into the solution and read the EC value. Reading was noted on pH meter when it was stabilized. The electrode was washed with distilled water and dried with tissue paper (vi) Determination of organic matter The empty crucible was weight (W1). The crucible was weight again when 5 g of compost sample was added in it (W2). The crucible having compost in it was placed in an oven at 105° C for 4 hours. After drying the sample, the crucibles were placed in muffle furnace at 800° C for 2 hours. The crucibles contain ash was weight (W3) the percentage of organic matter was determined by subtracting W1 from W2 and W1 fromW3. (vii) Determination of C: N The 0.5g of compost sample was taken and transferred in digestion tube.1.0g of digestion mixture was added in the compost sample. Then 10 to 12ml of sulphuric acid was also added in digestion tube. The digestion tube was placed in digestion block and heated for 2 hours at 400° C. The contents in the tube were changed color from black to light green. The digestion tube was cooled down. The sample was distillated on distillation apparatus in the presence of boric acid and sodium hydroxide. Carbon was divided by nitrogen to find the C: N. (viii) Enumerations of microflora in the vermicompost of E.eugeniae

Enumeration of microbial population was carried out in the vermicompost (vermicasts) by following the method suggested by Allen (1953). The population of bacteria, fungi and actinomycetes was determined in the substrate (vermicompost) of E.eugeniae. One gram of substrate was suspended in 99 mL of pre-sterilized distilled water in a 250 mL conical flask and kept in a mechanical shaker for 20 m at 120 rpm. Serial dilutions (10-3 to 10-6) were prepared using sterile distilled water. One mL of the aliquot of the 10-4 to 10-6 dilutions for bacteria, 10-3 to 10-4 dilutions for actinomycetes and fungus were pipetted out into sterile petridishes. Sterile Nutrient agar medium for bacteria, Glucose Asparagine agar medium for actinomycetes and Sabouraud’s Dextrose agar for fungus were prepared. About 15 to 20 mL of the above sterile media was poured at about 42 °C into the petriplates. The dishes were rotated clockwise and anticlockwise for uniform distribution of the samples. The plates were allowed to solidify and were incubated at 28  2 °C for 72 h. After incubation, the total number of colonies was counted in each plate. The population was expressed per gram of the substrate. Plating was performed in triplicates and average was recorded. 4. Results

The compost was analysed for physicochemical and biological parameters. The physico- chemical characteristics of the control substrate and worm worked substrate (experiment) are indicated in table 1. The pH was high in the substrate, the initial value of pH in the substrates S1 and S2, was recorded as 6.5, 7.15 respectively. The electrical conductivity was higher in the substrates in the initial day and found decreased on final day. But there was a drastic decline in organic carbon in substrates. At the end of the experiment the highest loss of organic carbon compounds. C: N ratio decreased from initial to final day of composting (Table 1). The substrates showed a significant increase in the percentage of NPK with the duration of 90 days.

Table -1 Chemical and physical characteristics of cattle dung and Vermicompost

Characteristics Cattle dung (S1) Vermicompost (S2) Initial Final Initial Final pH (1:10) 4.12 6.52 5.10 7.15 EC (1:10) (dS/m) 2.53 2.14 4.40 4.0

Bulk density (kg/m3) 58 42.00 1250 850.00 Moisture content (%) 20 11.00 70 65.00 Dry matter (%) 98 93.00 30 35.00 Organic matter (OM %) 100 97.50 95 63.10 Organic carbon (OC %) 65.2 51.71 45 25.00 Total nitrogen (TN %) 0.34 0.48 0.65 1.90

+ NH4 -N (mg/kg) 14.23 16.50 28.56 41.54

− NO3 -N (mg/kg) 5.54 7.75 8.89 10.67 C/N ratio 191.8 107.7:1 69.23:1 131.5:1 Ash (%) 0.61 0.50 65.0 56.90 Total phosphorus (%) 0.35 0.46 0.54 0.62 Total potassium (%) 0.65 0.79 0.39 0.55

Table -2 Microbial population of vermicompost.

Substrates /Days Bacteria X 106 cfu g-1 Fungi X 103 cfu g-1 Actinomycetes X 103cfu g-1

0 30 60 90 0 30 60 90 0 30 60 90

S1 -CD+Leaf litters 110 129 138 141 83 109 118 128 75 89 93 104 Without E.eugeniae

S2-CD+ Leaf litters 110 153 162 186 98 134 142 160 119 144 187 141 + E.eugeniae

CD- cow dung

Microbial population increased gradually from the initial day to final day of vermicomposting. Bacterial, fungal and actinomycetes population was found to be higher in the worm worked substrates than in the control substrates. On 90th day in worm worked substrate had maximum population of bacteria 186 X 106 cfu g-1, fungi 160 x 103 cfu g-1and actinomycetes 141 x 103 cfu g-1. We prepared the vermibricks using binding substances to retain the moisture and microbes.We used the vermibricks in the herbal garden as each brick for each plot. Use of this type of organic fertilizer therefore has great potential.

Discussion Principals of organic farming are to produce food of good quality and quantity by using eco-friendly technologies, which can co-exist with nature. Such practices exclude use of chemical fertilizers, pesticides and weedicides etc. The system depends upon use of organic manures (vermicompost) and microbial inoculations. The present study focused to produce vermicompost and the compost can be made as a brick to store easily to retain moisture and microbes. The compost was analysed for physicochemical and biological parameters. The results are supporting the earlier findings. The alkalinity of the substrate may be reduced to neutral during vermicomposting. The reduction of OC was due to the respiratory activity of earthworm and microorganisms. This type of reduction was also reported by Gunjal and Nikam (1992), Daniel and Anderson (1992). The decrease of EC in the vermicompost than the control showed that the activity of earthworm and microbes brought about the mineralization. Rao et al. (1978) and Karmegam et al. (1997) also reported similar observations. Higher microbial population in the worm casts of different earthworm species compared with underlying soil have been observed: bacteria and fungi in L. mauritii and E. eugeniae (Parthasarathi and Ranganathan, 1998) Since earthworms are soil residents they play an active role in improving the soil quality

Conclusion This project is completed to give user friendly and eco friendly product out of vermicompost. By using this vermibricks, the farmers can reduce the man power in the supplement of organic manure for their farms. According to their convenient the size and weight of the vermibricks can be modified. This is an industry which can be started without electricity, so it can be started without electricity hence it can be started in remote areas. Hence 50 % of chemical fertilizer can be replaced by this organic manure, so saving of power used for production of that quantity of chemical fertilizers. The soil becomes more porous so 50% irrigation water is saved, 50% electricity is saved,at the same time underground water table is maintained, erosion of top fertile soil is reduced and chances of flood is also reduced, and health of soil, plants, animals, labourers, farmers and consumers is maintained and less disease and major diseases like Cancer can be eradicated at the grass root level by reducing non protein nitrates and residual effects of pesticides from soil, plants and human beings.

References

1. Allen, G.N., 1953. Experiment in soil bacteriology. Burger’s Publication Co. Minnepolis, Minn., USA. 124. 2. Daniel, O., and J.M. Anderson, 1992. “Microbial biomass and activity in contrasting soil material after passage through the gut of the earthworm Lumbricus rubellus Hoffmeister.” Soil Biol. Biochem. Volume 24, pp. 465 - 470. 3. Daniel. T.,and N. Karmegam, 1999. Bio-conversion of selected leaf litters using an African epigeic earthworm, Eudrilus eugeniae. Ecol. Env. & Cons. 5: 271-275. 4. Gunjal, S.S. and T.B. Nikam, 1992. Grape cultivation through earthworm farming. In: Proc, of National Seminar on Organic Farming. Mahatama Phule Krishi Vidyapeeth, College of Agriculture, Pune, India, pp. 48-50. 5. Karmegam, N., G. Ananth, K. Alagumalai, and Daniel Thilagavathy, 1997. Utilization of organic substrates by vermiculture biotechnology for the growth of Raphanus salivus L. In: Nat. Sem. Sus. Env., N. Sukumaran, (Ed.), Sri Parama Kalyani Centre for Environmental Sciences. M.S. University.Alwarkurichi, Tamil Nadu, India. pp 57-58. 6. Jayashree. S., J.Rathinamala and P. Lakshmanaperumalsamy. 2008. Biocomposting of leaf litters by Eudrilus eugeniae and its application on the growth of green gram (Vigna radiata (L) CO.6). The Journ. of Solid Waste Technology and Management. 34 (2) 102- 112. 7. Mba, C.C. 1983. Utilization of Eudrilus eugeniae for disposal of cassava peel. In: Earthorm eology from Darwin to vermiculture (Eds.). J.E. Satchell) Chapman and Hall. London. pp. 315-321. 8. Parthasarathi, K and L.S. Ranganathan, 1998. Pressmud vermicasts are “hot spots” of fungi and bacteria. Ecology Environment and Conservation, 4(3): 81 – 86. 9. Ramalingam, R., 1997. Studies on the lifecycle, growth and population dynamics of Lampito mauritii (Kinberg) Eudrilus eugeniae (Kinberg) (Annelida: Oligochaeta) cultured in different organic wastes and analysis of nutrients and microbes of vermicomposts, Ph.D. Thesis, Annamalai University, India. 10. Rao, K.B., N. Ramarathanam, S.S. Prihar, and M.R. Quaser, 1978. “Changes in bulk density of soils due to changes inmoisture content.” J. Indian Soc.Soil Sci., Volume 26, pp.320-322.