Description:FIELD OF INVENTION

The present invention relates to microbial consortium for biodegradation of organic matter from agricultural waste. The present invention particularly relates to the microbial consortium comprising aerobic and/or facultative aerobic microbial population for the biodegradation of organic matter from any waste generated post-harvest under aerobic conditions.

BACKGROUND OF THE INVENTION

Biodegradation of agricultural waste such as leaves, soybean straw, wheat straw, and cotton stalks is a severe problem, so developing a plan to deal with these waste is critical. The majority of the time, these waste products are burned on the field; however this results in the emission of dangerous gases such as CO2, CO2, NO2, and other pollutants, as well as negative influence on the ozone layer. Composting allows these agricultural wastes to break down or biodegrade completely in a shorter amount of time. Fungi, in particular, can accelerate the decomposition process since they have the ability to consume wood. Aspergillus, Penicillium, and Trichoderma are fungal species that play an essential role in the degradation of diverse waste materials. Fungi are responsible for the decomposition of over 80% of cellulose. Remains of plants and animals in various stages of decomposition.

Composting reduces greenhouse gas emissions, particularly methane. Food waste in landfills makes up 14.1% of US methane emissions. Methane largely contributes to climate change by trapping heat at extremely high rates. NASA estimates methane was responsible for 23% of climate change in the 20th century, and 20% of that is due to poor waste management. Composting increases the amount of carbon stored in soil, otherwise known as creating a “carbon sink”. With more carbon being retained by soil, it significantly reduces greenhouse gas emissions and detrimental carbons in the atmosphere. Researchers Favoino and Hogg said, “An increase of just 0.15% in organic carbon in arable soils…would effectively imply the sequestration of the same amount of carbon within soil that is currently released into the atmosphere through the use of fossil fuels.” Composting captures carbon in soil which reduces its effects on climate change, an issue in desperate need of mediation.

Finally, composting has many other benefits for our agriculture and environment regeneration efforts. Compost minimizes our use of harsh chemical fertilizers and even promotes higher agricultural yields. By nourishing the soil rather than the plants themselves, compost promotes healthier, more abundant and nutritious plants, fruits, and vegetables. Compost can also revitalize soil that has been contaminated by hazardous waste; this is particularly helpful to environments that are densely populated. Additionally, compost increases water retention, and minimizes land degradation and soil erosion, which is helpful in farming, reforestation, and habitat revitalization efforts.

“Microbes run the world. It's that simple.” Microbial communities make the essential elements of oxygen, carbon, nitrogen, and sulphur available for other life on our planet. Without microbial decomposer communities, life would be smothered in dead organisms. Microbial consortia are a mixture of microorganisms isolated from the natural environment that works synergistically in hydrolysing biomass during anaerobic digestion. The species diversity of microbial consortia differs dramatically depending on their environmental conditions.

A microbial consortium enables microbes to use a broad range of carbon sources. It provides microbes with robustness in response to environmental stress factors. Microbes in a consortium can perform complex functions that are impossible for a single organism. With advancement of technology, it is now possible to understand microbial interaction mechanism and construct consortia. Microbial consortia can be classified in terms of their construction, modes of interaction, and functions.

Microbes can interact with each other in different ways, including commensalism and mutualism. Interactions between microbial communities decide the structure of consortia by enabling functions such as the division of labour, cross-feeding, and providing robustness against changing environments. However, considerable efforts are required to make this concept a reality.

Microbes that can interact (cross-talk) and support the growth of each other can be co-cultured. Their modes of interaction can be classified as synergistic, commensal, and mutualist.

Microbial consortia are designed to exhibit certain functions and support the growth of each other. It is an emerging field needing a more specific method of classification that can help us understand and construct a microbial consortium in an easy way.

IN202141027866 relates to sustainable materials like areca leaf that they are rarely composted and end up in a landfill. This invention related to the microbial composting of the used areca leaf materials for bioproducts such as enzymes, biofuels, potent microbial consortium for faster degradation of areca plates containing wastes and production of high CN containing compost manure for crops.

EP2550244B1 relates to a microbial consortium for treating organic waste. The present disclosure also relates to a composition, a system and a kit for treating organic waste. The first aspect of the invention, there is provided a process for treating organic waste, the process comprising the step of contacting an organic waste with one or more microorganisms.

US10358370B2 describes a method of processing municipal solid waste (MSW) comprising the steps of —providing a stream of unsorted MSW to a microbial fermentation reactor in which the MSW is fermented with agitation at a non-water content of between 10 and 50% by weight and at a temperature of between 35 and 75 degrees for a period of between 1 and 72 hours under conditions sufficient to maintain a live lactic acid bacteria concentration of at least 10.000.000.000 CFU/L, and —removing a stream of fermented unsorted MSW from the reactor and subjecting it to a separation step whereby non-degradable solids are removed to provide a slurry of bio-degradable components. Further, a slurry of bio-degradable components prepared according to said method is provided.

The microbial consortia in prior arts as mentioned above are limited to use only for municipal waste or organic household waste.

The microbial consortium of the present invention takes advantage over the prior arts by developing more effective microbial consortium for treatment of waste agricultural waste stubbles under anaerobic conditions.

SUMMARY OF THE INVENTION

The principle aspect of the present invention is to provide a microbe-based composition comprising a microorganism consortia including two or more microorganisms that are co-cultured in-situ, wherein the microorganisms of the consortia are selected from a group of: (i) microbe A is a class of cellulose degrading microbe; (ii) microbe B is a class of pectin degrading microbe; (iii) microbe C is a class of lignin degrading microbe; (iv) microbe D is a class of hemicellulose degrading microbe; (v) microbe E is a class of biofertilizer microbial strain; or (vi) microbe F is a class of biofungicide microbial strain.

In an aspect of the present invention, the microbe A is a class of cellulose degrading microbe selected from Cellulomonas cartae, Clostridium, Bacillus sp., Pseudomonas sp., Streptomyces, Fibrobacter succinogenes, Trichoderma sp., Penicillium, Aspergillus sp., Pseudo-lagarobasidium acaciicola and Nocardia sp.

In a further aspect of the present invention, the microbe B is a class of pectin degrading microbe selected from Bacillus subtilis, Bacillus licheniformis, Bacillus badius, Bacillus asahin, Bacillus psychrosaccharolyticus, Bacillus megaterium, Pseudomonas aeruginosa, Erwinia sp., Staphylococcus sp., Geobacillus sp, Anoxybacillus sp, Bacteroides, Magnaporthe oryzae, Giberella zeae, Botrytis fuckeliana, Sclerotinia sclerotiorum, Aspergillus nidulans, Trichoderma virens, Podospora anserine, Rhizopus oryzae, and Aspergillus clavatus.

In yet another aspect of the present invention, the microbe C is a class of lignin degrading microbe selected from Mycobacterium tuberculosum, M. avium, Pseudomonas syringae, P. aeruginosa, P. putida, Bordetella pertussis, Xanthomonas campestris, Escherichia coli, Caulobacter crescentus, Rhodobacter capsulatus, Yersinia pestis, Campylobacter jejuni, Aquifex aeolicus, Streptomyces antibioticus, S. griseus, S. coelicolor, S. cyaneus, S. lavendulae, Rhizoctonia solani, Aspergillus nidulans, Podospora anserina, Neurospora crassa, Gaeumannomyces graminis var. tritici, Trichoderma reesei, Hypholoma sp., Phanerochaete chrysosporium Pseudolagarobasidium acaciicola, Nematoloma sp., Pleurotus sp., and Armillaria sp.

In another aspect of the present invention, the microbe D is a class of hemicellulose degrading microbe selected from Alternaria solani, Botryosphaeria ribis, Botrytis allii, Corticium centrifugum, Monilia fructigena, Neurospora, Penicillium digitatum, Rhizopus nigricans, Sclerotinia fructigena, Gibberella saubinetti, Helminthosporium oryzae, Phytophthora infestans, Trametes gibbosa, Clostridium felsineum, Bacillus subtilis, Acetenobacter mannanolyticus, Bacillus aroideae, Sporocytophaga myxococcoides, Streptococcus sp., Butyrivibrio fibrisolvens, Ruminococcus albus, and Bacteroides ruminicola.

In a further aspect of the present invention, the microbe E is a class of bio-fertilizer microbe selected from a group of Rhizobium sp., Mesorhizobium sp., Azorhizobium sp., Bradyrhizobium sp., Sinorhizobium sp., Allorhizobium sp., Achromobacter sp., Alcaligenes sp., Arthrobacter, Acetobacter sp., Azomonas sp., Beijerinckia sp., Clostridium sp., Bacillus sp., Enterobacter sp., Erwinia sp., Desulfovibrio sp., Derxia sp., Corynebacterium sp., Campylobacter sp., Herbaspirillum sp., Klebsiella sp., Lignobacter sp., Mycobacterium sp., Rhodospirillum sp., Rhodo-pseudomonas sp., Xanthobacter sp., Methylosinus sp., Agrobacterium sp., Azotobacter sp., Acinetobacter sp., Actinoplanes sp., Frankia sp., Pseudomonas sp., Pleurotus ostreatus, Agaricomycetes sp., Micrococcus sp., Streptomyces sp., Xanthomonas sp., Enterobacter sp., Cellulomonas sp., Serratia sp., Flavobacterium sp., Thiobacillus sp., and Azospirillum sp.

In yet another aspect of the present invention, the microbe F is a bio-fungicide microbe selected from a group of Metschnikowia pulcherrima, Pichia membranifaciens, Wickerhamomyces anomalus, Meyerozyma guilliermondii, Aureobasidium pullulans, Aureobasidium subglaciale, Serratia plymuthica, Yamadazyma mexicana, Lactobacillus plantarum, Debaryomyces hansenii, Metschnikowia pulcherrima, Pichia guilliermondii, Rhodotorula minuta, Pichia kudriavzevii, Bacillus amyloliquefaciens, Paecilomyces lilacinus, Penicillium citrinum, Lactobacillus acidophilus, Streptomyces sp., Bacillus pumilus, Bacillus subtilis, Clavispora lusitaniae, Bacillus velezensis, Pseudomonas fluorescens, Streptomyces tuirus, Trichoderma viride, Trichoderma koningiopsis, Trichoderma asperellum, Trichoderma harzianum, Aureobasidium pullulans, Torulaspora globosa and Torulaspora indica.

In an aspect of the present invention, the microbial-based consortium is prepared on-site under conditions favourable to all species of microorganisms disclosed in this invention.

In an aspect of the present invention, the co-cultured microbial consortia does not require water to prepare a bio-composting culture.

In yet another aspect of the present invention, is to provide a microbe-based composition capable of degrading cellulose.

A further aspect of the present invention, is to provide a microbe-based composition capable of degrading pectin.

Another aspect of the present invention, is to provide a microbe-based composition capable of lignin degradation.

In yet another aspect of the present invention, a microbe-based composition is capable of degrading hemicellulose by producing enzymes.

A further aspect of the present invention, is to provide a microbe-based composition which is used as a biofertilizer.

Another aspect of the present invention is to provide a microbe-based composition which is used as a biofungicide.

A further aspect of the present invention is to provide a microbe-based composition which comprises of biological enzymes selected from cellulases, ligninases, pectinases, hemicellulases or combination thereof.

Another aspect of the present invention is to provide a microbe-based composition comprising of a carbon source.

Yet another aspect of the present invention is to provide a microbe-based composition applied to the dry stubble through the method of spraying.

A final aspect of the present invention is to provide a microbe-based composition capable of decomposing 72% to 85% of dry stubble by weight in 12 to 15 days.

The foregoing and other features of the disclosure will become more apparent from the following detailed description, which proceeds with reference to the accompanying figures.

BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 illustrates graphical representation of % cellulose degradation by the microbial consortia of the present invention at various locations.
Figure 2 illustrates graphical representation of % hemicellulose degradation by the microbial consortia of the present invention at various locations.
Figure 3 illustrates graphical representation of % lignin degradation by the microbial consortia of the present invention at various locations.
Figure 4 illustrates graphical representation of % increase of nutrients in the soil at Daun Kalan and Dhindhsa.
Figure 5 illustrates graphical representation of % increase of nutrients in the soil in the villages of Barnala District.
Figure 6 illustrates graphical representation of % increase of Carbon, Nitrate Nitrogen, Ammonical Nitrogen, Phosphorus and Potassium in the soil in the villages of Sangrur District.
Figure 7 illustrates graphical representation of % increase of Magnesium, Sulphur, Zinc, Boron and Iron in the soil in the villages of Sangrur District.
Figure 8 illustrates graphical representation of % increase of Magnesium, Sulphur, Zinc, Boron and Iron in the soil in the villages of Barnala District.

DETAILED DESCRIPTION OF THE INVENTION

A detailed description of the preferred embodiment of the present invention is disclosed herein; however, it is to be understood that the disclosed embodiment is merely exemplary of the invention, which may be embodied in various forms. Therefore, specific functional and structural details disclosed herein are not to be interpreted as limiting, but merely as a basis for the claims and as a representative basis for teaching one skilled in the art to employ the present invention in virtually any appropriately detailed structure. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention.

The present invention provides a ready to use microbial consortium. In general, microbial consortium is a mixture of two or more microbial species, which in some instances are in physical contact with one another. The microbes in a consortium may affect one another by direct physical contact or through biochemical interactions, or both.

The term "microbial consortia" or "microbial consortium" refers to a subset of a microbial community of individual microbial species, or strains of a species, which can be described as carrying out a common function, or can be described as participating in, or leading to, or correlating with, a recognizable parameter or plant phenotypic trait. The community may comprise two or more species, or strains of a species, of microbes. In some instances, the microbes coexist within the community symbiotically.

The term "microbial community" means a group of microbes comprising two or more species or strains. Unlike microbial consortia, a microbial community does not have to be carrying out a common function, or does not have to be participating in, or leading to, or correlating with, a recognizable parameter or plant phenotypic trait.

“In-situ” is a Latin phrase meaning 'in place' or 'on site', derived from in ('in') and situ (ablative of situs, lit. 'place'). The term refers to the examination or occurrence of a process within its original context, without relocation. The term is widely used across disciplines to denote methods, observations, or interventions carried out in their natural or intended environment. In situ methods have applications in diverse fields of applied science. In the aerospace industry, in situ inspection protocols and monitoring systems assess operational performance without disrupting functionality. Environmental science employs in situ ecosystem monitoring to collect accurate data without artificial interference. In medicine, particularly oncology, carcinoma in situ refers to early-stage cancers that remain confined to their point of origin. This classification, indicating no invasion of surrounding tissues, plays a crucial role in determining treatment plans and prognosis. Space exploration relies on in situ research methods to conduct direct observational studies and data collection on celestial bodies, avoiding the challenges of sample-return missions.

“Biomass” refers to cellulose- and/or starch-containing raw materials, including but not limited to wood chips, corn stover, rice, grasses, forages, perrie-grass, potatoes, tubers, roots, whole ground corn, grape pomace, cobs, grains, wheat, barley, rye, milo, brans, cereals, sugar-containing raw materials (e.g., molasses, fruit materials, sugar cane, or sugar beets), wood, and plant residues.

“Compost” is a mixture of ingredients used as plant fertilizer and to improve soil's physical, chemical, and biological properties. It is commonly prepared by decomposing plant and food waste, recycling organic materials, and manure. The resulting mixture is rich in plant nutrients and beneficial organisms, such as bacteria, protozoa, nematodes, and fungi. Compost improves soil fertility in gardens, landscaping, horticulture, urban agriculture, and organic farming, reducing dependency on commercial chemical fertilizers. The benefits of compost include providing nutrients to crops as fertilizer, acting as a soil conditioner, increasing the humus or humic acid contents of the soil, and introducing beneficial microbes that help to suppress pathogens in the soil and reduce soil-borne diseases.

At the simplest level, composting requires gathering a mix of green waste (nitrogen-rich materials such as leaves, grass, dry stubble, and food scraps) and brown waste (woody materials rich in carbon, such as stalks, paper, and wood chips). The materials break down into humus in a process taking months. Composting can be a multistep, closely monitored process with measured inputs of water, air, and carbon- and nitrogen-rich materials. The decomposition process is aided by shredding the plant matter, adding water, and ensuring proper aeration by regularly turning the mixture in a process using open piles or windrows. Fungi, bacteria, and other detritivores further break up the organic material. Aerobic bacteria and fungi manage the chemical process by converting the inputs into heat, carbon dioxide, and ammonium ions.

“Composting” is the natural process of recycling organic matter, such as leaves and food scraps, into a valuable fertilizer that can enrich soil and plants. Anything that grows decomposes eventually; composting simply speeds up the process by providing an ideal environment for bacteria, fungi, and other decomposing organisms (such as worms, sow bugs, and nematodes) to do their work. The resulting decomposed matter, which often ends up looking like fertile garden soil, is called compost. Fondly referred to by farmers as “black gold,” compost is rich in nutrients and can be used for gardening, horticulture, and agriculture.

“Biodegradation” is the breakdown of organic matter by microorganisms, such as bacteria and fungi. It is generally assumed to be a natural process, which differentiates it from composting. Composting is a human-driven process in which biodegradation occurs under a specific set of circumstances.

The process of biodegradation is threefold: first an object undergoes bio-deterioration, which is the mechanical weakening of its structure; then follows bio-fragmentation, which is the breakdown of materials by microorganisms; and finally assimilation, which is the incorporation of the old material into new cells.

In practice, almost all chemical compounds and materials are subject to biodegradation, the key element being time. Things like vegetables may degrade within days, while glass and some plastics take many millennia to decompose. A standard for biodegradability used by the European Union is that greater than 90% of the original material must be converted into CO2, water and minerals by biological processes within 6 months.

Plant extracts often contain a mixture of polysaccharides that can be conjugated with other bioactive and functional components, it is reasonable to summarize the most common polysaccharides found in plant materials. For this reason, the following topics will explore different types (e.g., fructans, hemicelluloses, and pectins), but also their effects on processes such as human inflammation, fibrosis, and metabolism.

Essentially, two different types of plant tissue reach the soil for decomposition: parenchymatic tissue and woody tissue. Parenchymatic cells are found in the living green tissue of leaves and in the cortex (bark) of young twigs and fine roots. They are composed of cellulose walls, the protoplast, rich in protein, and the vacuole. Woody tissues form the woody part (xylem) and the supporting tissue (sclerenchym) of stems leaf epidermis, leaf ribs and barks. The different layers of the woody cell wall (middle lamella, primary wall, secondary wall, tertiary wall) can be differentiated in their structure as well as in their chemical composition. The middle lamella, which acts as the binding substance between the cells consists of pectin and in woody tissues also of lignin. Primary wall, secondary wall, and tertiary wall consist of cellulose, polyoses (hemicelluloses) and lignin. The middle layer (primary wall and middle lamella) has the highest lignin concentration (40% to 60%). The largest part of the lignin is derived from the secondary wall, whose lignin concentration only amounts to 20-30%.

Plant tissues can be divided into various compound classes, including storage materials that are intracellular, and structural components that occur in membranes, extracellular or as cell wall constituents. The storage materials of plants are easily degradable and thus are important carbon and energy sources for microorganisms.

The major organic compounds of plant litter are polysaccharides and lignin. For example spruce needles are composed of 20% cellulose and lignin, 12% polyoses, 1-5% protein and 1-6% ash. Leaf litter contains 8-14% ash, 10-19% hemicelluloses, 10-22% cellulose, 5-8% lignin and 2-15% raw protein.

For example, dry banana leaves primarily consist of lignocellulose, which includes cellulose, hemicellulose and lignin. These components are found in varying percentages, with cellulose and hemicellulose being the most abundant. The leaves also contain other organic components like carbon and nitrogen, and the C/N ratio is typically high. Cellulose: The main structural component, providing rigidity and strength to the leaf. Hemicellulose: A complex carbohydrate that contributes to the structural integrity of the leaf. Lignin: A complex polymer that provides water resistance and support.

Since, major component of the plant tissue contains complex polysaccharides, breaking down these polysaccharides is a vital part of the decomposition process. Hence, microbes that are capable of degrading such complex sugars are used in the present invention to formulate a microbial consortia. Breaking down each of the complex polysaccharide compound is important to obtain the best result of biodegradation process.

Lignin Degradation is the process by which lignin, a complex plant-based polymer, is broken down into smaller molecules. This process can occur through biological or thermochemical means.

Pectin degradation is the breakdown of pectin, a complex polysaccharide found in plant cell walls. This process is catalysed by enzymes called pectinases. Pectinases, produced by various organisms including plants, microorganisms, and some animals, degrade pectin through different mechanisms, including hydrolysis and de-esterification.

Cellulose degradation is a complex process requiring a group of enzymes that synergize. Degradation can be done chemically or biochemically. Biochemical degradation has an advantage over the thermochemical degradation as it leads to the destruction of carbohydrates, whereas in biochemical degradation, the original carbohydrate moiety is preserved. Enzymes secreted from bacteria and fungi are involved in the degradation process of cellulose and are classified under the category of cellulase. New enzyme and enzyme modulators have been discovered that increase the efficiency of classical cellulase.

The classical cellulose degradation scheme requires three different enzyme systems produced by microbes that convert the polymeric substrate into its monomeric glucose. Endo-β-1,4-glucanases (endocellulases, EC 3.2.1.9.1), cellobiohydrolases (exocellulases, EC 3.2.1.91) and β-1,4-glucosidases (cellobiases, EC3.2.1.21). The alliance of these enzyme activities only enables them to interrupt the structure at the solid-liquid interface making the individual fibre available for hydrolysis. All three enzymes are hydrolases and cleave glycosidic bonds by the addition of water molecules.

Hemicellulose degradation is a process where hemicellulases, a type of enzyme can break down hemicellulose, which is a complex carbohydrate found in the cell walls of plants. Hemicellulases are produced by microorganisms such as bacteria and fungi, as well as by some animals and insects that consume plant material.
There are several different types of hemicellulases, each with a specific function and substrate specificity. For example, xylanases are hemicellulases that break down xylan, a type of hemicellulose found in some plant tissues. Similarly, mannanases can break down mannans, another type of hemicellulose.

Hemicellulases play an important role in the biodegradation of plant material in natural environments, as well as in industrial processes such as the production of biofuels and paper. They are also used in animal feed to improve the digestibility of plant-based feed ingredients.

In an embodiment of the present invention, the microbial-based composition includes co-cultured microorganisms comprising at least one microbe is selected from (A) a class of cellulose degrading microbes; microbe (B) a class of pectin degrading microbes; microbe (C) a class of lignin degrading microbes; microbe (D) a class of hemicellulose degrading microbes; microbe (E) a class of biofertilizer microbial strains; and microbe (F) a class of biofungicide microbial strains.

In another embodiment of the present invention, the microbial-based consortium is prepared on-site under conditions favourable to all species of microorganisms disclosed in this invention.

In yet another embodiment of the present invention, wherein the microbe (A) a class of cellulose degrading microbes is selected from Cellulomonas cartae, Clostridium, Bacillus sp., Pseudomonas sp., Streptomyces, Fibrobacter succinogenes, Trichoderma sp., Penicillium, Aspergillus sp., Pseudo-lagarobasidium acaciicola and Nocardia sp. in the amount of 5% to 30% w/w.

In a further embodiment of the present invention, wherein the microbe (B) pectin degrading microbe is selected from Bacillus subtilis, Bacillus licheniformis, Bacillus badius, Bacillus asahin, Bacillus psychrosaccharolyticus, Bacillus megaterium, Pseudomonas aeruginosa, Erwinia sp., Staphylococcus sp., Geobacillus sp, Anoxybacillus sp, Bacteroides, Magnaporthe oryzae, Giberella zeae, Botrytis fuckeliana, Sclerotinia sclerotiorum, Aspergillus nidulans, Trichoderma virens, Podospora anserine, Rhizopus oryzae, and Aspergillus clavatus in an amount of 7% to 40% w/w.

In yet another embodiment of the present invention, wherein the microbe (C) lignin degrading microbe is selected from Mycobacterium tuberculosum, M. avium, Pseudomonas syringae, P. aeruginosa, P. putida, Bordetella pertussis, Xanthomonas campestris, Escherichia coli, Caulobacter crescentus, Rhodobacter capsulatus, Yersinia pestis, Campylobacter jejuni, Aquifex aeolicus, Streptomyces antibioticus, S. griseus, S. coelicolor, S. cyaneus, S. lavendulae, Rhizoctonia solani, Aspergillus nidulans, Podospora anserina, Neurospora crassa, Gaeumannomyces graminis var. tritici, Trichoderma reesei, Hypholoma sp., Phanerochaete chrysosporium Pseudolagarobasidium acaciicola, Nematoloma sp., Pleurotus sp., and Armillaria sp. in an amount of 5% to 20% w/w.

In another embodiment of the present invention, wherein the microbe (D) hemicellulose degrading microbe is selected from Alternaria solani, Botryosphaeria ribis, Botrytis allii, Corticium centrifugum, Monilia fructigena, Neurospora, Penicillium digitatum, Rhizopus nigricans, Sclerotinia fructigena, Gibberella saubinetti, Helminthosporium oryzae, Phytophthora infestans, Trametes gibbosa, Clostridium felsineum, Bacillus subtilis, Acetenobacter mannanolyticus, Bacillus aroideae, Sporocytophaga myxococcoides, Streptococcus sp., Butyrivibrio fibrisolvens, Ruminococcus albus, and Bacteroides ruminicola in an amount of 3% to 20% w/w.

In a further embodiment of the present invention, wherein the microbe (E) bio-fertilizer microbes are selected from a group of Rhizobium sp., Mesorhizobium sp., Azorhizobium sp., Bradyrhizobium sp., Sinorhizobium sp., Allorhizobium sp., Achromobacter sp., Alcaligenes sp., Arthrobacter, Acetobacter sp., Azomonas sp., Beijerinckia sp., Clostridium sp., Bacillus sp., Enterobacter sp., Erwinia sp., Desulfovibrio sp., Derxia sp., Corynebacterium sp., Campylobacter sp., Herbaspirillum sp., Klebsiella sp., Lignobacter sp., Mycobacterium sp., Rhodospirillum sp., Rhodo-pseudomonas sp., Xanthobacter sp., Methylosinus sp., Agrobacterium sp., Azotobacter sp., Acinetobacter sp., Actinoplanes sp., Frankia sp., Pseudomonas sp., Pleurotus ostreatus, Agaricomycetes sp., Micrococcus sp., Streptomyces sp., Xanthomonas sp., Enterobacter sp., Cellulomonas sp., Serratia sp., Flavobacterium sp., Thiobacillus sp., and Azospirillum sp. in an amount of 5% to 30% w/w.

In yet another embodiment of the present invention, wherein the microbe (F) bio-fungicide microbes are selected from a group of Metschnikowia pulcherrima, Pichia membranifaciens, Wickerhamomyces anomalus, Meyerozyma guilliermondii, Aureobasidium pullulans, Aureobasidium subglaciale, Serratia plymuthica, Yamadazyma mexicana, Lactobacillus plantarum, Debaryomyces hansenii, Metschnikowia pulcherrima, Pichia guilliermondii, Rhodotorula minuta, Pichia kudriavzevii, Bacillus amyloliquefaciens, Paecilomyces lilacinus, Penicillium citrinum, Lactobacillus acidophilus, Streptomyces sp., Bacillus pumilus, Bacillus subtilis, Clavispora lusitaniae, Bacillus velezensis, Pseudomonas fluorescens, Streptomyces tuirus, Trichoderma viride, Trichoderma koningiopsis, Trichoderma asperellum, Trichoderma harzianum, Aureobasidium pullulans, Torulaspora globosa and Torulaspora indica in an amount of 5% to 30% w/w.

A further embodiment of the present invention is to provide cellulase, hemicellulases, pectinases and ligninase enzymes for improved degradation of cellulose, hemicellulose, pectin and lignin, respectively.

Another embodiment of the present invention is to provide a carbon source for the consortium in the in-situ co-culturing stage.

Yet another embodiment of the present invention is to provide the microbial consortium dry condition to decompose agricultural waste stubble from broadleaf and grass crops.

Another embodiment of the present invention is to provide the agricultural waste stubble is selected from the group comprising banana, cassava, cereals, citrus, cocoas, flowers, fibre crops, forge corps, grapes, groundnuts, hops, oil palm, oilseed rape, peas, potato, rice, sugarcane, sunflower, tea, tobacco, wheat, and corn.

A further embodiment of the present invention is to prepare a formulations of microbial composition selected from Wettable Powder (WP), Water-Dispersible Granule (WDG), Flowables (F), Emulsifiable Concentrates (EC), Suspension Concentrates (SC) and aerosol formulation.

In a further embodiment of the present invention is to mix the microbial composition with inactive excipients.

In yet another embodiment of the present invention, the inactive excipients include surfactants, buffer, dispersant, wetting agent, disintegrating agent, solvent and diluent.

In a further embodiment of the present invention, the surfactant compound is selected from Sodium Lauryl Sulfate (SLS), Sodium Laureth Sulfate (SLES), Ammonium Lauryl Sulfate, Sodium Stearate, Alcohol Ethoxylates, Cocamide, Decyl Glucoside, Alkyl Polyglucoside, Betaines, Silicone Surfactants and amino oxides.

In yet another embodiment of the present invention, the solvent is selected from Demineralized water (DM water), brines, sugar-rich aqueous milieu, saline suspension, and Deep eutectic solvents.

In another embodiment of the present invention, the diluents used are selected from Distilled water, 0.9% saline solution, Bentonite, montmorillonite, and kaolin.

In yet another embodiment of the present invention, the wetting agent is selected from polyethylene, polypropylene glycol

The final embodiment of the present invention is to return the decomposed organic waste to the soil to improve the nutrient composition of the soil post-harvest.

Further, the following examples illustrate the present invention:

Example 1: Co-culturing microorganisms for preparation of microbe-based cellulose and pectin degrading consortium

The microbes were inoculated in water consisting of glycerol as the carbon source. The inoculum of each microbe is 10% of the total volume of the culture. The pH of the culture medium as adjusted to pH 7. To this mixture, biological enzymes were added such as pectinase and cellulases. The resultant composition is mixed well at room temperature.

Table 1 (a): In situ Biocomposting Culture 1 (BC-1)
Ingredients Percentages (%) share in formulation (mL)
Bacillus megaterium 20
Cellulomonas cartae 20
Rhodopseudomonas sp. 20
Pseudo-lagarobasidium acaciicola 20
Bacillus licheniformis 20
Cellulase enzyme (powder in gram) 0.5
Pectinase enzyme (powder in gram) 0.5
Glycerol 0.5
Total microbial inoculum percentages (%) 100

Example 2: Co-culturing microorganisms for preparation of microbe-based Cellulose and lignin degrading consortium

The microbes were inoculated in water consisting of glycerol as the carbon source. The inoculum of each microbe is 10% of the total volume of the culture. The pH of the culture medium as adjusted to pH 7. To this mixture, biological enzymes were added such as ligninases and cellulases. The resultant composition is mixed well at room temperature.

Table 2: In situ Biocomposting Culture 2 (BC-2)
Ingredients Percentages (%) share in formulation (mL)
Trichoderma viride 7-10
Paecilomyces lilacinus 7-10
Pleurotus ostreatus 7-10
Phanerochaete chrysosporium 7-10
Pseudolagarobasidium acaciicola 7-10
Agaricomycetes sp. 7-10
Torulaspora globosa 7-10
Saccharomyces cerevisiae 7-10
Bacillus megaterium 7-10
Cellulomonas cartae 7-10
Rhodopseudomonas sp. 7-10
Cellulase enzyme (powder in gram) 0.5
Ligninase enzyme (powder in gram) 0.5
Glycerol 0.5
Total microbial inoculum percentages (%) 100

Example 3: Co-culturing microorganisms for preparation of microbe-based hemicellulose and pectin degrading consortium

The microbes were inoculated in water consisting of glycerol as the carbon source. The inoculum of each microbe is 10% of the total volume of the culture. The pH of the culture medium as adjusted to pH 7. To this mixture, biological enzymes were added such as pectinase and hemicellulases. The resultant composition is mixed well at room temperature.

Table 3: In situ Biocomposting Culture 3 (BC-3)
Ingredients Percentages (%) share in formulation (mL)
Botrytis fuckeliana 10
Aspergillus clavatus 40
Rhizopus nigricans 05
Pleurotus ostreatus 15
Trichoderma viride 30
Hemicellulase enzyme (powder in gram) 0.5
Pectinase enzyme (powder in gram) 0.5
Glycerol 0.5
Total microbial inoculum percentages (%) 100

Example 4: Co-culturing microorganisms for preparation of microbe-based consortium without enzyme supplementation

The microbes were inoculated in water consisting of glycerol as the carbon source. The inoculum of each microbe is 10% of the total volume of the culture. The pH of the culture medium as adjusted to pH 7. The resultant composition is mixed well at room temperature.

Table 4: In situ Biocomposting Culture 4 (BC-4)
Ingredients Percentages (%) share in formulation (mL)
Trichoderma viride 7-10
Paecilomyces lilacinus 7-10
Pleurotus ostreatus 7-10
Phanerochaete chrysosporium 7-10
Pseudolagarobasidium acaciicola 7-10
Agaricomycetes sp. 7-10
Torulaspora globosa 7-10
Saccharomyces cerevisiae 7-10
Bacillus megaterium 7-10
Cellulomonas cartae 7-10
Rhodopseudomonas sp. 7-10
Glycerol 0.5
Total microbial inoculum percentages (%) 100

Example 5: Comparative field analysis of the microbial consortia

All 4 microbial cultures were analysed in the field on dry stubble waste to determine the efficacy of the microbial consortia. The microbial consortia of the present invention outperformed the other three consortia in terms of efficacy, synergism and % waste degradation.

Table 5: Comparative study of Biocomposting culture of the present invention
Microbial Consortium Days for stubble degradation Dry stubble waste degradation (%)
Biocomposting Culture 1 (Bacteria-only consortium) 18-20 50-55%
Biocomposting Culture 2 (Present invention) 12-15 72-85%
Biocomposting Culture 3 (Fungus-only Consortium) 14-17 65-70%
Biocomposting Culture 4 (Enzyme-less Consortium) 20-25 45-55%

Example 6: Preparing sprayable microbial-based composition

4 Litres of microbial-based composition is mixed in 200 Litres at atmospheric temperature. Subsequently, 4 litres of the resultant mixture is sprayed per acre of agricultural waste stubbles. The process of spraying is carried out by the use of pressure pumps or garden sprinklers which further reduces labour time and cost.

Example 7: Biodegradation of stubble waste

After spraying the said mixture into the agriculture field, the soil and the stubbles are mixed together to maintain moisture. The mixture of stubble and soil is left in the open for 12-15 days. The microbe-based composition is provided with aerobic condition as it is kept in open air. Incubation period of 12-15 days is under atmospheric temperature and conditions. At the end of the incubation period, polysaccharides such as Cellulose, Hemicellulose, Pectin and Lignin are degraded by 60% to 85%.

Table 7: Evaluation of cellulose, Hemicellulose and Lignin degradation
Microbial Culture Incubation Period
Degradation %

Cellulose Hemicellulose Lignin
Microbial Consortium LTF-27 [Zheng et al. 2020] 20 days 71.70 65.60
12.50

Microbial Consortium DN-1 [Xu et al. 2015]
46 days
62.57 67.14 42.54
Streptomyces griseorubens C-5 [Xu & Yang 2010]
45 days
43.88 49.86 58.47
IARI PUSA Tablets 20-25 days 82.65 61.55 55.56
Commercially available products 25-30 days 62.58 58.75 41.02
Present Invention 12-15 days 83.78 62.55 68.75

Example 8: Replenishing Soil nutrients
The decomposed stubble waste is a rich source of organic fertilizer compounds. Stubble waste from rice straws are broken down into basic soil elements such as carbon, Nitrogen, Phosphorus, Potassium etc. In addition to this, the soil bacterial and fungal count is also improved as a result of biodegradation of the said waste stubble.

Table 8: Evaluation % increase of nutrients in soil
Parameters Average Increase in % Least increase in % Most increase in %
Organic Carbon 63.7 17.5 144.0
Nitrate Nitrogen 162.2 37.7 371.5
Ammonical Nitrogen 136.5 62.8 318.3
Phosphorus 146.6 23.5 408.5
Potassium 81.0 32.7 153.3
Magnesium 69.6 32.9 126.9
Sulphur 84.5 24.3 398.2
Zinc 99.8 19.8 400.0
Boron 110.3 15.0 233.3
Iron 139.4 38.6 401.1
Bacterial Count 21.0 times 12.4 times 33.6 times
Fungal Count 22.6 times 10.2 times 45.0 times

Conclusion: the microbial consortium increases the general nutrient content of the soil through biodegradation of organic waste. In addition to improving the nutrient bioavailability to the crops grown in the field, it also improves the microorganism content in the soil.
, C , C , C , Claims:CLAIMS:
I/We Claim:
1. A microbe-based composition for biodegradation of waste stubble, comprising of:
a. a microorganism consortia including two or more microorganisms which are co-cultured in-situ, and the microorganisms in the consortia are selected from a group of:
i. microbe (A) a cellulose degrading microbe in an amount of 5% to 30% w/w;
ii. microbe (B) a pectin degrading microbe in an amount of 7% to 40% w/w;
iii. microbe (C) a lignin degrading microbe in an amount of 5% to 20% w/w;
iv. microbe (D) a hemicellulose degrading microbe 3% to 20% w/w;
v. microbe (E) a biofertilizer microbial strain in an amount of 5% to 30% w/w; and
vi. microbe (F) a biofungicide microbial strain in an amount of 5% to 30% w/w;
b. biological enzymes selected from cellulases, ligninases, hemicellulases, pectinases or combination thereof in an amount of 0.1 to 0.5 g/ml;
c. a carbon source in an amount of 0.1 to 0.5 g/ml; and
d. formulation excipients.

2. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein the microbial consortia was prepared comprising multiple microbial strain selected from each of the microbe (A to F) which are mixed together to form a multispecies consortia.

3. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein the microbe strains exist in the consortium through symbiotic interaction with each of the other microbial strains of the consortia.

4. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein a cellulose degrading microbial strain is selected from Cellulomonas cartae, Clostridium, Bacillus sp., Pseudomonas sp., Streptomyces, Fibrobacter succinogenes, Trichoderma sp., Penicillium, Aspergillus sp., Pseudo-lagarobasidium acaciicola and Nocardia sp.

5. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein a pectin degrading microbe species is selected from Bacillus subtilis, Bacillus licheniformis, Bacillus badius, Bacillus asahin, Bacillus psychrosaccharolyticus, Bacillus megaterium, Pseudomonas aeruginosa, Erwinia sp., Staphylococcus sp., Geobacillus sp, Anoxybacillus sp, Bacteroides, Magnaporthe oryzae, Giberella zeae, Botrytis fuckeliana, Sclerotinia sclerotiorum, Aspergillus nidulans, Trichoderma virens, Podospora anserine, Rhizopus oryzae, and Aspergillus clavatus.

6. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein one or more lignin degrading microbes are selected from Mycobacterium tuberculosum, M. avium, Pseudomonas syringae, P. aeruginosa, P. putida, Bordetella pertussis, Xanthomonas campestris, Escherichia coli, Caulobacter crescentus, Rhodobacter capsulatus, Yersinia pestis, Campylobacter jejuni, Aquifex aeolicus, Streptomyces antibioticus, S. griseus, S. coelicolor, S. cyaneus, S. lavendulae, Rhizoctonia solani, Aspergillus nidulans, Podospora anserina, Neurospora crassa, Gaeumannomyces graminis var. tritici, Trichoderma reesei, Hypholoma sp., Phanerochaete chrysosporium Pseudolagarobasidium acaciicola, Nematoloma sp., Pleurotus sp., and Armillaria sp.

7. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein a hemicellulose degrading microbe is selected from Alternaria solani, Botryosphaeria ribis, Botrytis allii, Corticium centrifugum, Monilia fructigena, Neurospora, Penicillium digitatum, Rhizopus nigricans, Sclerotinia fructigena, Gibberella saubinetti, Helminthosporium oryzae, Phytophthora infestans, Trametes gibbosa, Clostridium felsineum, Bacillus subtilis, Acetenobacter mannanolyticus, Bacillus aroideae, Sporocytophaga myxococcoides, Streptococcus sp., Butyrivibrio fibrisolvens, Ruminococcus albus, and Bacteroides ruminicola.

8. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein one or more bio-fertilizer microbes are selected from a group of Rhizobium sp., Mesorhizobium sp., Azorhizobium sp., Bradyrhizobium sp., Sinorhizobium sp., Allorhizobium sp., Achromobacter sp., Alcaligenes sp., Arthrobacter, Acetobacter sp., Azomonas sp., Beijerinckia sp., Clostridium sp., Bacillus sp., Enterobacter sp., Erwinia sp., Desulfovibrio sp., Derxia sp., Corynebacterium sp., Campylobacter sp., Herbaspirillum sp., Klebsiella sp., Lignobacter sp., Mycobacterium sp., Rhodospirillum sp., Rhodo-pseudomonas sp., Xanthobacter sp., Methylosinus sp., Agrobacterium sp., Azotobacter sp., Acinetobacter sp., Actinoplanes sp., Frankia sp., Pseudomonas sp., Pleurotus ostreatus, Agaricomycetes sp., Micrococcus sp., Streptomyces sp., Xanthomonas sp., Enterobacter sp., Cellulomonas sp., Serratia sp., Flavobacterium sp., Thiobacillus sp., and Azospirillum sp..

9. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein one or more bio-fungicide microbes are selected from a group of Metschnikowia pulcherrima, Pichia membranifaciens, Wickerhamomyces anomalus, Meyerozyma guilliermondii, Aureobasidium pullulans, Aureobasidium subglaciale, Serratia plymuthica, Yamadazyma mexicana, Lactobacillus plantarum, Debaryomyces hansenii, Metschnikowia pulcherrima, Pichia guilliermondii, Rhodotorula minuta, Pichia kudriavzevii, Bacillus amyloliquefaciens, Paecilomyces lilacinus, Penicillium citrinum, Lactobacillus acidophilus, Streptomyces sp., Bacillus pumilus, Bacillus subtilis, Clavispora lusitaniae, Bacillus velezensis, Pseudomonas fluorescens, Streptomyces tuirus, Trichoderma viride, Trichoderma koningiopsis, Trichoderma asperellum, Trichoderma harzianum, Aureobasidium pullulans, Torulaspora globosa and Torulaspora indica.

10. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein the biological enzymes are selected from cellulases, hemicellulases, pectinases, ligninases or combination thereof.

11. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein the carbon source is selected from ethanol, formate, lactate, butyrate, glycerol, acetic acid, fructose, maltose, whey protein, waste stubble and complex polysaccharides like starch, cellulose or a mixture thereof.

12. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein the microbial consortia degrades polysaccharides like cellulose, pectin, lignin and hemicellulose.

13. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein the microbial consortia improves nutrient content of the soil.

14. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein 2% to 3% w/w of microbial consortia is mixed with 200 L v/v of water at 35 ̊ C to 40 ̊ C to prepare a sprayable microbial composition.

15. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein the waste stubble is degraded over a period of 12 to 15 days after mixing with microbial composition.

16. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein the waste stubble takes no less than 12 days to degrade 72% to 85% by weight.

17. The microbe-based consortia for biodegradation of waste stubble as claimed in claim 1, wherein the waste stubble takes no more than 15 days to degrade 72% to 85% by weight.

18. The microbe-based consortia as claimed in claim 1, wherein the degraded waste stubble is a natural fertilizer.

Dated this 19th Day of June 2025.