TITLE:
In-situ aerobic rapid composting method of agricultural organic waste
FIELD OF INVENTION:
This invention relates to a composition and method for on-farm [in-situ] composting of agricultural organic wastes to useful organic manure and release of nutrients
OBJECT OF THE INVENTION:
The principal object of this invention is to provide methodology for treating and/or composting agricultural waste to organic manure and release of nutrients. Second object of the invention relates to selection of microbes for treating and/or composting agricultural waste.
Third object of the invention is to provide a composition of selected microbes for rapid conversation of agricultural waste into organic manure and release of nutrients in an environment friendly manner.
BACKGROUND OF THE INVENTION:
Agricultural output has expanded by more than three times since last 50 years as a result of farm expansion for agricultural use, high yielding seeds, fertilizers and technological contributions from the green revolution. As a result of the growing production of agro-based products, there is increased agro related organic waste leading to accumulation and pollution of the environment. Harvest waste, also known as crop residue, includes both field residues that remain in an agricultural field or orchard after the crop has been harvested and process residues that remain after the crop has been processed into a useful component. Field leftovers include stalks and stubble (stems), leaves, and seed pods, to name a few. Substantial portion of agricultural waste [around 65-70%] generally accumulates and/ or are incinerated causing soil, air and water pollution.

India is one of the world's biggest agro-based economies, with major cultivation of paddy and wheat. High demand for food, fibre and feed has necessitated a steady increase in crop production with less available time in between cropping cycle, forcing the farmers to burn the crop residues for field preparation/sowing of the next crop. India is also one of the world's second-largest contributor of Crop residue burning (84 Tg/year) (Grover and Chaudhry, 2019). Farmers prefer crop residue burning (CRB) because it takes less time, is less expensive, and prepares the field for the following crop faster. In India, the overall amount of agricultural waste generated each year is substantially more than in other countries. Biomass burning is a major source of particulate matter (PM) and trace gases in the atmosphere, which has an impact on local and worldwide climates and poses serious health risks to humans. Furthermore, CRB emits a significant quantity of air pollutants including as nitrogen oxides (NOx), sulphur dioxide (SO2), carbon monoxide (CO), volatile organic compounds (VOCs), and particulate matter (PM), all of which have an impact on air quality. The nutrients [macro and micro nutrients, organic carbon, etc.,] present on the crop residue is lost on burning.
Agro-waste includes waste from agricultural products, agro-industries, animal feed, horticulture, aquaculture, and other sources. Due to the high expense of pre-processing processes, most agricultural wastes are abandoned or used inefficiently, posing considerable environment and safety problems. There are efforts for environmental friendly sustainable methods for waste management such as WO2021154088A1 relates to a process of producing soil amendment from organic waste with biochar, IN360956 claimed anaerobic biodegradation of agricultural waste with high C:N ratio, WO2016198798A1 discloses a dry-state anaerobic digestion method comprising pretreatment of the input with an additive selected from among zeolite, clay, pozzolan and biochar, WO2016077577A1 provides a method comprises anaerobically incubating a particulate additive in contact with a carbonaceous feedstock in a neutral or alkaline aqueous culture medium containing a culture of methanogenic consortia and collecting methane generated, WO2017210609A1 relates to the use of porous carbonaceous

structures in particular to biochar infused with microbial additives to enable or enhance the solubility of minerals that are beneficial for both plant and animal health and growth, WO2007060683A1 relates to a novel process for liquefaction of solid organic matter hydrolysis using anaerobically treated water containing a consortium of fermentative micro-organisms, WO2011019871A1 relates to a material comprises a biochar impregnated with a methanogen or algae, fungi, bacteria, archaea, protist, and their combination, WO2017117314A1 describes biochar as a delivery system for microbes and the microbes are infused in the biochar using a surfactant, WO2017139510A1 describes a method and system for using biochar in composting environments to control temperature, moisture, acid and pH levels, odors and bacterial cultures, CN113135777 relates on-site harmless treatment method for the fruit and vegetable waste. Therefore, it is of prime importance and one of the key challenges in agriculture to explore for cost-efficient, rapid, and sustainable alternative methods for the efficient management of agricultural organic waste on farm.
The introduction of an easy, cost effective and rapid handling technology to deal with solid organic waste is considered a basis of any country's sustainable agriculture. Using microorganisms to manage agricultural waste could be an effective solution for soil cleansing and pollution reduction. Composting is one such alternate to burning and other means of waste disposal. It provides several benefits over conventional management techniques, including minimal technical complexity and low cost, minor environmental impact because pathogens are normally eliminated during the high temperature phase of aerobic composting. The basic idea underlying composting is to use naturally existing microorganisms to accelerate the breakdown of organic waste. Microbial populations reduce the biomass's complex compounds into simpler ones that can be reused or recycled through natural processes. Microbial degradation processes lower soil toxicity, enhance plant growth by supplying growth-promoting metabolites, and give plant nutrients by sequestering minerals from the soil. During composting, organic

matter is biologically decomposed under aerobic conditions, making this procedure both environmentally benign and cost-effective. Because of its various positive effects on the physical and chemical properties of the soil, organic matter biodegradation contributes to reducing waste volume and producing a stable and nutrient-rich final end product, "compost," that can be used in agricultural activities while reducing the use of inorganic fertilizers.
The present invention provides an affordable composition and efficient in-situ composting method for converting complex organic materials into stable, humus¬like compounds.
The agricultural waste generated after harvesting crops, are burned in the field
causing very high emission of green-house gases, contamination, and pollution
especially in India currently. There is a need to prevent air pollution due to
burning of agricultural waste. There is also a need for an effective method and
composition of degradation of agricultural waste into a useful component without
causing any pollution. One of the many technologies used to manage agricultural
waste is composting which degrade complex organic substrate. Lignocellulose is
the predominant organic substance found in agricultural waste and is a highly
complex carbon source. It comprises of two carbohydrate polymers, cellulose and
hemicellulose, as well as the non-carbohydrate phenolic polymer, lignin.
Microorganisms have a rapid metabolism and fast reproduction rate. They have
the ability to produce variety of enzymes that can act on and break down
complicated substrates like lignocellulose. Using microorganisms with specific
enzyme activities helps accelerate the composting process. Microbial
communities produce enzymes that can break down lignocellulose. By adding microbial inoculum with specific functions to compost, one can artificially encourage the transformation of organic materials into stable humus. Extensive research has been conducted on aerobic composting, and the findings indicate to it as a potential sustainable, eco-friendly method for managing lignocellulosic agricultural waste and improving agriculture. This organic bioprocess enhances

agriculture's long-term sustainability by transforming agricultural waste into nutrient-rich compost.
By being inoculated with specific microbial cultures, agricultural waste can undergo aerobic fermentation, which accelerates the biological decomposition of the waste and enhances the quality of the compost. Composting or aerobic fermentation are both apparently identical. The aerobic fermentation process of organic waste differs from traditional composting in that it uses efficient microorganisms for the decomposition of organic waste.
The use of aerobic fermentation has the potential to produce high-quality compost from a large volume of useful agricultural waste. Under the optimal conditions of aeration, oxygen supply, temperature, moisture content, pH value, C:N, particle size, etc., aerobic microorganisms can oxidize small molecule organic matter in fermentation substrate and release the energy needed for biological growth activities through their life activities.
In most cases, adding microbial inoculum could considerably reduce the composting period, achieve higher temperatures, reduce offensive odor, and even improve the compost quality because inoculation could create advantageous microbial communities, improve microbiological quality, increase microbial population, and produce various desired enzymes. Furthermore, efficient microorganism could convert harmful and complex chemicals into non-hazardous and simpler molecules. The addition of microbial agents enhances the essential nutrient values in compost, the temperature profile during composting, and the diversity of the microbial community as a whole. A diverse range of microorganisms is responsible for breaking down the organic matter during the composting process, and different microorganisms are found at various phases of the composting process. Bacillus has emerged amongst several functional microorganisms that can be inoculated because of its favoured cellulase-producing ability, heat tolerance, and superior environmental adaption. Additionally, it has proven to be quite effective at composting agricultural waste. The disclosed embodiments provide a method for composting and/or treating agricultural waste wherein the method includes culturing bacteria in a nutrient

medium to obtain a microbial composition and further devising a formulation comprising of microbial inoculum and inert carrier elements.
SUMMARY:
The present invention discloses an effective microbial composition for in-situ aerobic biodegradation of agricultural waste and method of use of this composition. The microbes were isolated from Dumpsites/landfills which are normally considered of as storehouse of useful microorganisms that can thrive on organic waste.
The composition contains a consortium of beneficial Bacillus strains in organic carrier base that is used to accelerate and manage aerobic composting. These strains are capable of degrading long chain biopolymers (such as lignin, lignocelluloses, hemicelluloses, etc.) in a well-aerated environment, thereby speeding up the degradation of organic waste that would otherwise take more time by natural or conventional means. The agricultural waste mentioned here are pressmud, rice straw, cotton plant stalks, wheat straw, bamboo stalks, sugarcane trash or any other agricultural waste. The time for degradation of agricultural waste is 3 to 20 days, but preferably the process is completed by 21 days. The stable end-product is a compost which can be used in agriculture as a fertilizer or soil enhancer.
The bacterial strains have been deposited with International Depository Authority at Microbial Type Culture Collection, Chandigarh.
Example 1
Screening of microorganisms
A soil sample was taken from local dumpsite/landfills at Ahmedabad, Gujarat, INDIA. The sample was sufficiently diluted to produce well-isolated colonies on agar surface. Two bacterial isolates with distinct morphologies predominated the agar surface. The ability of each bacterial isolate to break down substrate such as cellulose, pectin, xylan, starch, lipid, lignin, etc. were examined, because agricultural waste is a rich source of cellulose, hemicellulose, starch, lipids, and

proteins. The enzyme activities were also determined at different temperatures (room temperature, 40°C and 50°C) and pH range (5, 6, 7 and 8) to assess their usability in variety of environmental conditions. These isolates were further identified as Bacillus cabrialesii and Bacillus mucilaginosus using 16S rDNA V3-V4 amplicon sequence analysis. Compatibility tests of these isolates were carried out prior to formulation design.
Example 2
Composition for conversion of agricultural waste to compost material or soil
enhancer
The composition of the present invention for conversion of agricultural waste to soil enhancer comprises a formulation of Bacillus cabrialesii and Bacillus mucilaginosus
Microbial inoculum formulation:
Microbial inoculum comprises of Bacillus species (MTCC 25570 and MTCC 25571). Bacillus species were grown in Nutrient broth medium at 300C for 48-72 hours with continuous shaking. The bacterial cells enter the sporulation stage during this prolonged incubation. This is then utilised to create Solid Concentrate Culture. For each kilogram of formulation, 0.05 kilogram Solid Concentrate Culture was mixed with lignite, rock phosphate, zeolite and silica powder (0.5, 0.1, 0.1 and 0.1 kilogram, respectively) and 0.15 litre of water. The resulting formulation was a ready to use dry solid powder.
Lignite helps to maintain the optimal moisture content required for survival of microorganisms, thereby improving the product shelf life. Zeolite is a microporous, crystalline aluminosilicate with large surface area and high chemical as well as thermal stability. It provides minerals like calcium and potassium and enhances water holding capacity. Silica helps in product compaction. Rock phosphate is an essential supply of phosphorus for microorganisms to carry out a variety of metabolic processes, including respiration, photosynthesis, ATP production, protein synthesis, and the synthesis of nucleic acids.

Note:
The bacterial strains have been deposited at International Depository Authority at Microbial Type Culture Collection, Chandigarh with an accession date 4th August 2022 and accession number MTCC 25570 and MTCC 25571 respectively. Bacillus species: It has been filed to National Biodiversity Authority File # 2522 dated 10th Sep 2018.
Example 3
An in-situ method of conversion of agricultural waste to compost material or
soil enhancer
The current method relates to in-situ aerobic composting of agricultural waste. 50 to 100 kg of organic manure was mixed with 1 Kg of the microbial formulation. Manure is basically an organic matter derived from animal, human and plant waste after composting, that is rich in plant nutrients in a complex organic form. Typically, the formulation is combined with the enriched composted manure before being broadcasted onto the field. This practice makes sure that the formulation is applied evenly and gives the microorganisms time to adapt and proliferate before they interact with the agricultural waste and decompose it. Then, this mixture is applied to an about one-acre freshly harvested field such as paddy, wheat field, and so on. The field is periodically sprinkled with water via sprinkler or flooded with water to maintain the appropriate moisture level. A rotavator is used to plough the land after 2–3 days of application. Again, occasional water sprinkling is performed to maintain the optimum moisture level and microbial activity. About 15 to 16 days after application, the area is ploughed all over again. These recurrent ploughing activities ensures mixing and aeration of soil. After 21 days, the field becomes ready for the subsequent crop. All of the agricultural waste is composted through the microbial activity, and the rich compost returns all the nutrients to the soil while maintaining the soil’s carbon content. This integrated system that tackles the agricultural residue problem by composting with microbial inoculum and subsequent field incorporation is

gaining worldwide attention. This eliminates the need for waste collection and transportation, which reduces the subsequent charges and labor costs. In-situ composting of agricultural waste can also be seen as a way to optimize the use of previously applied synthetic fertilizers that would otherwise be lost through incineration or would have end up lying in landfills.
Example 4
A method for aerobic heap composting of agricultural waste:
The formulation of example 2 can be used multiple ways. It can be used for aerobic heap composting of agricultural waste. The process commences by shredding the waste into small pieces approximately 4 to 6 inches. This is mixed with a formulation containing a microbial inoculum. For every tonne of organic waste, 1 kg of formulation is used. Based on the initial moisture content of the waste, the formulation is dissolved in 10-15 ltrs of water and broadcasted evenly onto 1 tonne of organic waste. If the organic waste has high moisture content, the product can be directly applied. Following mixing, the material is shaped into a heap about 5 to 6 feet high and placed loosely. Optimum aeration, moisture and temperature are maintained throughout the process. The heap height will decrease by 20% to 25% within three days. This indicates that the heap is properly established. Regular rotation of the pile maintains optimum oxygen levels. It is very important to monitor the oxygen percentage throughout the process. Oxygen levels are kept above 10% to avoid anaerobic pockets in the heap. Moisture content is maintained between 50-55% for efficient composting. A low initial moisture content will dehydrate the pile and stop microbial growth. High humidity, on the other hand, makes microorganisms insignificant to the composting process and creates anaerobic conditions that cause the formation of foul-smelling greenhouse gases. Temperature monitoring is critical throughout the process to ensure pathogen removal while maintaining ideal conditions for the microbial community. Exothermic biological activity of aerobic bacteria raises temperature to 65-70°C in a few days of composting. Carbon to nitrogen (C: N)

ratio is kept below 40 during composting process. The proportions of carbon and nitrogen in composting materials have specific significance. C: N ratio is not only vital for composting but also impacts the quality of end product. If it is too high, the decomposition may take a long time and additionally nitrogenous material may be required to bring it down to the acceptable level. At the end of the process, the C: N Ratio is kept below 15. The first turn is done on the 3rd day. The first 1feet of the heap is placed inside while the remaining compost is placed outside. The process is continued 8th, 13th day and 18th day. Finally, the material is spread for curing on 21st day.
REFERENCES
Grover, D., Chaudhry, S. Ambient air quality changes after stubble burning in
rice–wheat system in an agricultural state of India. Environ Sci Pollut Res 26,
20550–20559 (2019). https://doi.org/10.1007/s11356-019-05395-5
WO2021154088A1: A process of producing soil amendment from organic waste, and a fertilizer produced from same
IN360956: Inoculant for rapid biodegradation of agricultural waste and method of use
WO2016198798A1: New solid-state methanation process
WO2016077577A1: Method for generating methane from a carbonaceous feedstock
WO2017210609A1: Mineral solubilizing microorganism infused biochars
WO2007060683A1: A microbial consortium and use thereof for liquefaction of solid organic matter
WO2011019871A2: Biochars, methods of using biochars, methods of making biochars, and reactors
WO2017117314: Biochar as a microbial carrier
WO2017139510: Biochars for use in composting
CN113135777: Fruit and vegetable waste in-situ harmless treatment method

CLAIMS
1. A composition for in-situ aerobic composting by biodegradation of an agricultural waste comprises microbial inoculum of Bacillus species with accession numbers MTCC 25570 and MTCC 25571
2. The composition of claim 1 further comprises inert carrier elements such as lignite, zeolite, silica powder, rock phosphate
3. An in- situ aerobic composting method for rapidly composting agricultural waste into organic manure using the composition of claim 1 that improves soil nutrient quality and serve as an ideal alternative to crop residue burning.
4. A method of selection of microbes for the composition of claim 1 for optimum biodegradation of an agricultural waste