Description:FIELD OF THE INVENTION
Present invention relates to field of biofertilizer and more particularly relates to composition and method for preparation of a biogas slurry based biofertilizer comprising biogas slurry, bio stimulant, microbes, enzymes and herbal extract as a source of organic nutrient. The biofertilizer improves soil quality and thereby soil fertility by nitrogen fixation, phosphorus solubilization, potassium mobilization, nutrient up taking, soil structure enhancement, disease suppression, environment sustainability, stress control and biological nutrient fixation. Use of the biofertilizer replaces chemical fertilizers which are harmful for the soil, water and environment. The biofertilizer is cost effective, ecofriendly and adaptable to a wide range of crops.
BACKGROUND OF THE INVENTION
With the increase in population and change in demographics, availability of adequate amount of food and agricultural products is going to be a major challenge in the selected geographical locations of the developing and underdeveloped countries.
Among the various methods, use of higher amount of chemical fertilizers for increasing the agricultural production results into environmental pollution, persistent changes in the soil ecology, physiochemical composition, decrease in agricultural productivity and health hazards. Various climatic factors enhance abiotic stress on crops resulting in reduced agricultural productivity. Various abiotic stress factors include soil salinity, drought, wind, improper temperature, heavy metals, waterlogging, and different weeds. Various biotic stress factors include phytopathogens like bacteria, viruses, fungi, and nematodes that attack plants, reducing crop productivity and quality.
Because of the various drawbacks associated with the use of chemical fertilizer and to reduce or overcome the effect of various stress factors on crop, use of the biofertilizers which provide nutrition through natural processes like zinc, potassium and phosphorus solubilization, nitrogen fixation, production of hormones, siderophore, various hydrolytic enzymes and protect the plant from different plant pathogens and stress conditions is becoming more popular. The biofertilizer with custom composition can provide the nutrition in adequate amount that is sufficient for healthy crop development, eco-friendly and economically convenient.
Utilizing biogas slurry for agricultural production is most suitable method out of various methods of disposal of biogas slurry. Biogas slurry is superior to other available organic fertilizers prepared by heap retting methods in terms of the material cycle, nutrient richness, and organic matter contents (Chemosphere, Volume 234, November 2019, Pages 953-961; New Biotechnology, Volume 61, 25 March 2021, Pages 80-89). Theoretically, the biogas slurry is suitable for growing crops. However, the content of mineral elements in the biogas slurry is uneven, in which the contents of N and K are too much high, but the contents of P, Ca, and Mg are abnormally low, restricting the agricultural utilization of biogas slurry. The biogas slurry contains a high concentration of mineral elements, including ammonia nitrogen (NH4+-N) and potassium (K) that can easily cause the secondary pollution to the environment once they are not appropriately handled. Therefore, with the increases in the number of biogas plant and hence the amount of biogas slurry produced across the globe, biogas slurry disposal is crucial and needs special attention (Science of The Total Environment, Volume 890, 10 September 2023, 164383).
Various methods for biogas slurry disposal include: (i) natural biological disposal, (ii) biochemical disposal and (iii) utilizing the biogas slurry for agricultural production. In natural biological disposal method, pollutants such as ammonia, nitrogen, phosphorus, and chemical oxygen demand (COD) from the biogas slurry are removed using the hydroponic cash crops in the biological ponds or constructed wetlands. This method of disposal occupies large area, emits odour and sufferers from groundwater contamination risk. (New Biotechnology, Volume 61, 25 March 2021, Pages 80-89). Biochemical disposal method features high treatment efficiency. But as a complex method it requires involving the processes such as anoxic/oxic (A/O), sequencing batch reactor (SBR), sequencing biofilm batch reactor (SBBR), and their joint processes. This method features a heavy maintenance workload, a significant investment, high energy consumption and, poor resource recycling.
DESCRIPTION OF THE RELATED ART
Various biofertilizer compositions and their method of preparation, mechanisms of action, role in crop productivity and in biotic/abiotic stress tolerance is reported.
Atmospheric condition specific and constituent specific biofertilizer formulations are disclosed. Patent publication no. US8415271B2 discloses biofertilizer formulations that contain one or more microorganisms capable of solubilizing phosphorus at low temperatures, specifically starting from 0° C. Patent publication no. CN107445674A discloses a production system of a selenium-enriched biofertilizer.
Biofertilizer formulations preparation from the specific raw material is disclosed. Patent publication no. CN101928184A discloses an organic biofertilizer comprises of human manures, poultry manures and animal slurry, waste material slags and crop straws serving as raw materials along with an additive and a preparation method thereof. Patent application no. US2004/0031302A1 discloses an integrated method to utilize to a maximum extent the fresh biomass of seaweeds such as Kappaphycus alvarezii that can be crushed to release sap and where the sap is useful as a potent liquid fertilizer after suitable treatment with additives and dilution while the residue is a superior raw material for extraction of -carrageenan, thereby enhancing the value of the seaweed. Patent application no. US 2021/0331987 A1 discloses processes for manufacturing compositions for plants and soils, such as liquid biofertilizers and solid bio stimulants, from animal manure. Patent application no. CN105384534A discloses composition and method for preparation of liquid organic biofertilizer from mud and natural pond liquid.
Biofertilizer formulations to be used in combination with other fertilizers are disclosed. Patent publication no. WO2014042517A2 discloses a biofertilizer to be used in combination with an NKP fertilizer, comprising at least one bacterial strain selected from the group comprising Pseudomonas aeruginosa strain AGKT 1, Serratia marcescens strain AG T4 and Bacillus amyloliquefaciens capable of solubilizing P in a liquid medium at the amount of at least 100 µg/mL.
Plant specific biofertilizers are disclosed. Patent publication no. CN104151065A discloses biogas slurry based long-acting biofertilizer for watermelons.
OBJECT OF THE INVENTION
Principal object of present invention is to contribute to sustainable agricultural practices by providing a nutrient-rich and environmentally friendly alternative to chemical fertilizers in the form of biogas slurry based biofertilizer that utilize biogas slurry and organic waste effectively, turning waste into a valuable resource for agriculture, thereby contributing to waste management and resource recycling.
Another object of the present invention is to provide a biofertilizer that is convenient to use, ensuring that farmers can integrate it seamlessly into their existing agricultural practices.
Another object of present invention is to provide a biofertilizer that is environment friendly and reduces carbon footprint associated with agriculture practices and minimize adverse effects on soil and water quality.
Another object of present invention is to provide a biofertilizer that efficiently extract and deliver essential nutrients from biogas slurry and fermented organic manure to plants, ensuring optimal nutrient availability for growth and easy up taken of the nutrients by plant.
Another objective of present invention is to provide a biofertilizer containing herbal extract that provides immunity to plants to fight against diseases and pests and also allow them to survive in stressed condition.
Another object of present invention is to provide a biofertilizer that enhance soil structure, microbial diversity, and overall soil health, promoting a balanced and thriving soil ecosystem.
Another object of present invention is to provide a biofertilizer that increases soil fertility and provides sustainable agriculture, and global efforts to address environmental challenges associated with conventional farming practices.
Another object of the present invention is to provide a biofertilizer that improve crop productivity by delivering a well-balanced formulation of nutrients essential for plant growth, leading to increased yields.
Another object of the present invention is to provide biofertilizer that protect crop by providing tolerance against salinity, drought, or other climatic impacts.
Further object of the present invention is to provide a biofertilizer and a method for preparation that improve crop productivity by delivering a well-balanced formulation of nutrients essential for plant growth, leading to increased yields.
Biogas slurry bases biofertilizer as per present invention provides immunity to plants, help plant to get proper nutrient, improve soil health, increase soil fertility, waste utilization and increase crop yield simultaneously addressing problem of biogas slurry disposal.
Usage of chemical fertilizer provides higher crop yield and is economical but harmful to the environment. Use of chemical fertilizer degrades soil texture, soil fertility, and chemical runoff to water bodies during monsoons. As all the products for farming to grow their crop are available separately and they are not feasible for farmer to buy it like organic manure, bio stimulant, biofertilizer and herbal extract are available separately. Purchasing multiple products like organic manure, bio stimulant, biofertilizer and herbal extract increases the cost of farming. The digested biogas slurry remaining after production of biogas is quite difficult to handle as it is the main source of soil contamination and harmful for the environment.
The biogas slurry based biofertilizer composition as per present invention includes biogas slurry, water, carbon source, buffer, herbal extract, Calcium source, inoculant, additive and stimulant increase crop yield, economical, nourish soil, improve soil texture and fertility, provide plants with proper nutrients and immunity to fight disease, and are 100% organic, which doesn’t harm the environment.
The biogas slurry is rich in organic nutrient like nitrogen, phosphorous, potassium and micronutrients. Biogas slurry adds organic matters to the soil, improving soil structure water retention and promoting microbial activity.
SUMMARY OF THE INVENTION
A biogas slurry based biofertilizer composition according to present invention is comprises of 40-70% by weight of biogas slurry, 30-60% by weight of water, 0.08-0.09% by weight of Jaggery, 0.1% by weight of Na2CO3, 0.08-0.09% by weight of KH2PO4, 0.8-0.9% by weight of herbal extract, 0.003-0.004% by weight of CaCO3, 8-9% by weight of an inoculum, 0.24-0.27% by weight of an additive and 0.32-0.44% by weight of a stimulant.
The herbal extract of the said biofertilizer composition is a decoction containing secondary metabolite of blend of a blend of 4.85% by weight of fenugreek, 0.16% by weight of turmeric, 0.16% by weight of asafoetida, 0.05% by weight of neem oil, 0.016% by weight of humic acid, 0.004% by weight of fulvic acid, 0.003% by weight of calcified seaweed, 0.16% by weight of cinnamon, 0.7% by weight of a micronutrient powder, 2.42% by weight of moringa powder and 3.23% by weight of aloe vera in 87.34% by weight of water. The micronutrient powder which is the part of herbal extract is consisting of 30.9% weight by weight (W/W) of Iron, 37.03% W/W of Zinc, 12.34% W/W of Copper, 12.34% W/W of Manganese, 1.23% W/W of Molybdenum and 6.2% W/W of Boron by Atomic Absorption Spectroscopy (AAS).
The inoculum is consisting of 108-109 CFU/mL of Azospirillum, 108-109 CFU/mL of Rhizobium, 108-109 CFU/mL of Azotobacter, 108-109 CFU/mL of Pseudomonas fluorescens, 108-109 CFU/mL of Bacillus subtilis, 108-109 CFU/mL of Bacillus megaterium and, 106-107 spores/gram of Mycorrhizae sp or any combination thereof.
The additive includes 0.08-0.09% by weight of Polysorbate 20, 0.08-0.09% by weight of Carboxymethyl cellulose (CMC) and 0.08-0.09% by weight of Polyethylene Glycol (PEG).
The stimulant is consisting of 0.2-0.3% by weight of Humic acid, 0.07-0.08% by weight of Fulvic acid and 0.05-0.06% by weight of Calcified seaweed.
A method for preparation of the biogas slurry based biofertilizer composition includes multistage fermentation process in which biogas slurry is first fermented anaerobically and then aerobically. Detailed steps are depicted in flow diagram shown in FIG. 1.
BRIEF DESCRIPTION OF THE DRAWINGS
The accompanying drawings provides a clear understanding of the present invention along with a detailed description, and it constitute a part of this complete specification.
FIG. 1 illustrates a flow diagram depicting steps for preparation of a biogas slurry based biofertilizer.
DETAILED DESCRIPTION OF THE INVENTION
The preferred embodiment describes a biogas slurry based biofertilizer composition and method of its preparation in detail with the help of FIG. 1.
Moreover, the detailed description of the present invention provides a clear understanding of the proportion and function of each individual constituent of the biogas slurry based biofertilizer composition.
Composition of the biogas slurry based biofertilizer
Present biofertilizer composition is an advance biofertilizer formulation made up of biogas slurry, water, a carbon source, buffers, a herbal extract, a Calcium source, an inoculum having beneficial microbes and enzymes, an additive and a stimulant. Because of interaction and chemical reaction between individual constituents of this biofertilizer resulting formulation has a synergetic effect on plant growth, maintenance and improvement of soil quality, reduction in effect of stress factors on plant growth and soil quality and reduction in environment pollution.
Present biofertilizer composition is consisting of 40-70% by weight of biogas slurry, 30-60% by weight of water, 0.08-0.09% by weight of Jaggery, 0.1% by weight of Na2CO3, 0.08-0.09% by weight of KH2PO4, 0.8-0.9% by weight of a herbal extract, 0.003-0.004% by weight of CaCO3, 8-9% by weight of the inoculum, 0.24-0.27% by weight of the additive and 0.32-0.44% by weight of a bio stimulant.
The herbal extract is a decoction containing secondary metabolite of blend of a blend of fenugreek, turmeric, asafoetida, neem oil, humic acid, fulvic acid, calcified seaweed, cinnamon, micronutrient, moringa powder and aloe vera in water. Following Table 1 shows proportion of each constituent in the blend used for preparing the decoction. Table 2 shows composition of micronutrient powder.
Table 1

Constituent Weight percent
Fenugreek 5.24%
Aloe vera 3.49%
Cinnamon 0.17%
Turmeric 0.17%
Moringa powder 2.6%
Neem oil 0.05%
Asafoetida 0.17%
Humic acid 0.017%
Fulvic acid 0.004%
Micronutrient powder 0.7%
Calcified seaweed 0.003%
Water 87.34%

Table 2
Constituent Weight by Weight (W/W) % as per *AAS
Iron 30.9
Zinc 37.03
Copper 12.34
Manganese 12.34
Molybdenum 1.23
Boron 6.2
*Atomic Absorption Spectroscopy
The inoculum is consisting microorganisms namely Azospirillum, Rhizobium, Azotobacter, Pseudomonas fluorescens, Bacillus subtilis, Bacillus megaterium and Mycorrhizae sp. Following Table 3 shows proportion of each constituent in the inoculum along with growth medium used. Growth medium is used to support the growth of a population of microorganisms or cells via the process of cell proliferation or small plants like the moss Physcomitrella patens.
Table 3
Constituent Colonogenic cell numbers Growth medium
Azospirillum 108 to 109 CFU/mL Nitrogen-free Bromothymol blue agar medium
Rhizobium 108 to 109 CFU/mL Yeast Extract Mannitol agar medium
Azotobacter 108 to 109 CFU/mL Bruk’s agar medium
Pseudomonas fluorescens 108 to 109 CFU/mL King’s B agar medium
Bacillus subtilis 108 to 109 CFU/mL Nutrient agar medium
Bacillus megaterium 108 to 109 CFU/mL Nutrient agar medium
Mycorrhizae sp 106 to 107 spores/gram Modified Melin-Norkrans agar Medium

The additive is consisting of Polysorbate 20, Carboxymethyl cellulose (CMC) and Polyethylene Glycol (PEG). Following Table 4 shows proportion of each constituents of the additive.
Table 4
Constituent Weight percent
Polysorbate 20 0.08-0.09%
Carboxymethyl cellulose (CMC) 0.08-0.09%
Polyethylene Glycol (PEG) 0.08-0.09%

The bio stimulant is consisting of 0.2-0.3% by weight of Humic acid, 0.07-0.08% by weight of Fulvic acid and 0.05-0.06% by weight of Calcified seaweed. Following Table 5 shows proportion of each constituents of the stimulant.
Table 5
Constituent Weight percent
Humic acid 0.2-0.3
Fulvic acid 0.07-0.08
Calcified seaweed 0.05-0.06

Biogas slurry is a nutrient-rich byproduct of anaerobic digestion. It serves as an organic fertilizer, providing essential nutrients like nitrogen, phosphorus, and potassium for plant growth. Biogas slurry is used as a base or carrier in the biofertilizers composition and hence it is named as biogas slurry based biofertilizer. It enriches the biofertilizer composition with organic nutrients, enhancing soil fertility.
Jaggery is used as a carbon source for growth of beneficial microbes.
A buffered pH is a necessity of most enzymes to function efficiently and correctly. Certain compounds alter soil pH. Na2CO3 and KH2PO4 are used as buffer solutions. When Na2CO3 is dissolved in liquid water, Na2CO3 is a salt that, when taken up, will raise the pH. KH2PO4, reduces the leachate pH.
Herbal extract contains bioactive compound that have antimicrobial or growth promoting property. Antimicrobial activity helps to suppress the pathogenic microbes on plants and in soil and enhance disease resistance in plants. While the formulation itself consist of beneficial microbes, which are adaptive to herbal extract and had become resistant towards antimicrobial agents present in the formulation due to anaerobic and aerobic fermentation process. Herbal extract contains secondary metabolite of fenugreek, turmeric, asafetida, neem oil, cinnamon, moringa and aloe vera.
Fenugreek seeds are rich in saponins, which have insecticidal and antibacterial qualities. By encouraging microbial activity in the soil, they improve nutrient availability and work with soil microbes to prevent the growth of pathogenic organisms. Quercetin and kaempferol are two flavonoids found in fenugreek seeds. The flavonoids have anti-inflammatory and antioxidant characteristics. These substances have the ability to affect soil microbial communities, encouraging the development of beneficial microorganisms involved in the cycling of nutrients and the stimulation of plant growth.
Turmeric includes compounds called curcuminoids, which have antibacterial, anti-inflammatory, and antioxidant qualities. By preventing the growth of pathogenic microorganisms and encouraging the growth of beneficial soil bacteria and fungi, these substances alter the microbial communities in the soil. One of the antimicrobial and insecticidal compounds found in turmeric is ar-turmerone or triterpenoids. By reducing the number of soil-borne pathogens and promoting the development of beneficial microbes, they have an impact on the microbial communities in the soil.
Asafoetida is a source of coumarins, such as umbelliferone, which possess antibacterial qualities. Through their ability to suppress the growth of pathogenic microbes and encourage the growth of beneficial soil bacteria and fungi, these substances affect the microbial communities in the soil. Asafoetida is a source of organosulfur compounds, such as ferulic acid, which have antibacterial qualities. They work in concert with soil microbes to promote the growth of beneficial soil bacteria and fungi while regulating the population of soil-borne pathogens.
Terpenoids, limonoids, and azadirachtin are secondary metabolites of neem. Strong insecticidal, antifungal, and antibacterial characteristics are displayed by these secondary metabolites of neem, which help to manage pests and soil-borne infections while promoting healthy microorganisms in biofertilizers.
Secondary metabolites of cinnamon are cinnamonaldehyde, cinnamonic acid, and phenolic compounds. The antibacterial and antioxidant characteristics of secondary metabolites of cinnamon are well-known, and they aid in the management of soil-borne infections as well as the stimulation of advantageous microbial activity in biofertilizers.
Glucosinolates are a compound found in moringa leaves that serves as a precursor to bioactive substances such as isothiocyanates. Because they change the makeup of soil fungi and bacteria and encourage the growth of beneficial microbes, these substances have an impact on the microbial communities in the soil. Moringa leaves contain phenolic compounds with antioxidant and antimicrobial properties. These substances have the ability to interact with soil microorganisms in a way that inhibits the growth of pathogenic organisms and encourages the growth of beneficial soil fungi and bacteria.
Aloe vera contains anthraquinones with antimicrobial and anti-inflammatory properties, such as emodin and aloin. These substances have the ability to affect the microbial communities in soil by preventing the growth of pathogenic microorganisms and encouraging the growth of beneficial soil bacteria. Aloe vera gel contains polysaccharides such as acemannan, which function as prebiotics to support the growth and activity of beneficial soil microbes in the rhizosphere.
Calcium carbonate (CaCO3), the chief component of limestone, is used to neutralize soil acidity and to supply calcium (Ca) for plant nutrition. Here, the term “lime” refers to ground limestone.
Beneficial microbes, such as nitrogen-fixing bacteria, phosphate-solubilizing bacteria, and mycorrhizal fungi, form symbiotic relationships with plants, improving nutrient availability. The application of microbes is a key component of biofertilizers. They contribute to nutrient cycling, fix atmospheric nitrogen, and enhance nutrient absorption by plants.
Beneficial microbes produce the active enzymes which catalysis the rate of reaction between organic matter and microbes and make nutrient more accessible to plants by breaking down of the organic matter. Enzymes are included in biofertilizer formulations to enhance the breakdown of organic residues in the soil, releasing nutrients for plant uptake.
Enzymes produced by Azospirillum, Rhizobium, Azotobacter, Pseudomonas fluorescens, Bacillus subtilis, Bacillus megaterium and Mycorrhizae sp which are constituents of the inoculum is consisting discussed below.
Azospirillum produces nitrogenase and nitrate reductase. Nitrogenase converts atmospheric nitrogen (N2) into ammonia (NH3) through nitrogen fixation. Nitrate reductase converts nitrate (NO3) into nitrite (NO2) and ultimately into ammonia (NH3).
Rhizobium produces nitrogenase, which is responsible for nitrogen fixation, converting atmospheric nitrogen (N2) into ammonia (NH3) within the root nodules of leguminous plants.
Azotobacter produces nitrogenase that facilitates nitrogen fixation, converting atmospheric nitrogen (N2) into ammonia (NH3) directly in the soil.
Pseudomonas fluorescens produces chitinase and proteases. Chitinase degrades chitin, a major component of fungal cell walls, contributing to the biological control of plant pathogens. Proteases are enzymes that hydrolyze proteins into simpler compounds, aiding in nutrient release and soil fertility.
Bacillus subtilis produces phosphatases, amylases, and cellulases. Phosphatases hydrolyze organic phosphates into inorganic phosphates, making phosphorus more available to plants. Amylases break down starch into simpler sugars, facilitating nutrient uptake by plants. Cellulases degrade cellulose, a component of plant cell walls, releasing nutrients for plant growth.
Bacillus megaterium produces phytase and lipases. Phytase catalyzes the hydrolysis of phytic acid, releasing phosphate from organic phosphorus compounds, thereby improving phosphorus availability to plants. Lipases break down lipids into fatty acids and glycerol, contributing to nutrient cycling in the soil.
Mycorrhizae produce phosphatase and chitinase. Phosphatase helps in the mineralization of organic phosphorus compounds, making phosphorus more accessible to plants. Chitinase is involved in the degradation of fungal cell walls, aiding in the suppression of soil-borne pathogens.
The efficiency of biofertilizers in improving soil fertility and encouraging plant growth is largely due to the vital roles these enzymes play in nitrogen fixation, nutrient cycling, and the biological control of plant pathogens.
Interaction of secondary metabolite with pathogenic and beneficial microbes
Plant secondary metabolites protect beneficial microbes in the soil while preventing the growth of pathogenic bacteria through a number of mechanisms, including:
Targeting Specific Types of Pathogens: Secondary metabolites frequently show selective antimicrobial activity, affecting only certain pathogen types and largely sparing beneficial microbes. The biochemical pathways or cell structures that separate pathogenic and helpful microbes account for this selectivity.
Secondary metabolites have the ability to obstruct pathogenic bacteria's vital biochemical functions, including DNA replication, protein synthesis, and cell wall synthesis. Beneficial microbes have different biochemical pathways or enzymes that make them less susceptible to the effects of these metabolites.
Microbial Competition: Some secondary metabolites give good bacteria an advantage over their harmful competitors by preventing their growth. For example, antimicrobial compounds released by plants suppress the growth of pathogenic fungi or bacteria, allowing beneficial microbes to thrive in their absence.
Indirect Effects: By altering the soil environment, secondary metabolites also indirectly encourage the growth of beneficial microorganisms. For example, certain compounds alter soil pH, organic matter content, or nutrient availability in ways that favour beneficial microbial populations over pathogens.
Microbial Adaptation: Beneficial microbes possess mechanisms to detoxify or metabolize secondary metabolites produced by plants. These adaptations allow them to coexist with the plant and even benefit from the compounds released by their host.
When herbal mixtures are mixed with biofertilizers, these plant-derived secondary metabolites contribute to the following effects on plant growth and microbial growth:
• enhanced soil fertility and nutrient availability,
• reduction in soil-borne diseases caused by pathogenic microbes,
• promotion of beneficial microbial populations involved in nutrient cycling and plant health and,
• improved plant energy, growth, and resilience to environmental stresses.
All these elements are usually put together to achieve a very synergistic result. Biofertilizer is formulated to consist of beneficial microbes in a carrier, such as biogas slurry, along with bio stimulants, enzymes, and herbal extracts to provide an integrated approach for plant nutrition and health.
Bio stimulant stimulate plant physiological processes, promote growth, and enhance stress tolerance. Applications of bio stimulant in biofertilizer formulations enhance the plant's response to microbial inoculants and improve nutrient uptake. Humic acid as a stimulant improves soil structure and nutrient retention. Fulvic acid as a stimulant enhances nutrient uptake and stress resistance in plants. Calcified seaweed as a stimulant helps to maintain pH of soil.
Polysorbate 20 as an additive acts as emulsifier/surfactant. Carboxymethyl cellulose (CMC) as an additive acts as binder/adjuvant. Polyethylene Glycol (PEG) as an additive acts as cell protectant.
Following Table 6-8 discloses the source and geographical origin of the biological material used in the present invention.
Table 6
Constituent Source and geographical origin
Azospirillum Isolated from soil collected from B.R.C garden, Udhna, Surat-394210, Gujarat, India
Rhizobium
Mycorrhizae sp
Pseudomonas fluorescens Isolated from cow dung obtained from BAPS Swami Narayan Temple, Nr. Sardar Bridge, Lalji Nagar Society, Adajan Gam, Adajan, Surat-395009, Gujarat, India
Bacillus subtilis
Bacillus megaterium
Azotobacter

Table 7
Constituent Source and Geographical origin
Fenugreek Purchased from local market, Surat, Gujarat, India
turmeric Purchased from local market, Surat, Gujarat, India
Asafoetida Purchased from local market, Surat, Gujarat, India
Neem oil Purchased from local market, Surat, Gujarat, India
Humic acid Purchased from local market, Jalgaon, Maharashtra, India
Fulvic acid Purchased from local market, Jalgaon, Maharashtra, India
Calcified seaweed Purchased from local market, Jalgaon, Maharashtra, India
Cinnamon Purchased from local market, Surat, Gujarat, India
Moringa powder Purchased from local market, Surat, Gujarat, India
Aloe vera Purchased from local market, Surat, Gujarat, India

Table 8
Constituent Source and Geographical origin/ Make
Biogas slurry Biofics anaerobic digester
Pandesara GIDC, Surat, Gujarat
Polysorbate 20 Commercial market Jalgaon Maharashtra
Carboxymethyl cellulose (CMC) Commercial market Jalgaon Maharashtra
Polyethylene Glycol (PEG) Commercial market Jalgaon Maharashtra
Jaggery Commercial market Surat Gujarat
CaCO3 Loba chemie
Na2CO3 Loba chemie
KH2PO4 Loba chemie
Inoculum Biofics R&D lab

Method for preparation of the biogas slurry based biofertilizer
FIG. 1 shows steps for preparation of a biogas slurry based biofertilizer. The biogas slurry based biofertilizer is prepared by a multistage fermentation process in which the biogas slurry is first fermented anaerobically and then aerobically along with beneficial microbes, bio stimulants, enzymes, and herbal extracts.
The decoction containing secondary metabolite is prepared by boiling blend of fenugreek, turmeric, asafoetida, calcium, neem, humic acid, fulvic acid, calcified seaweed, cinnamon, micronutrient, moringa and aloe vera in proportion as per Table 1 in water at 120 ºC for 2-4.
A mixture of 40-70 weight % of the biogas slurry, the 0.8-0.9 weight % of decoction of herbs as per Table 1, 0.32-0.44 weight % of bio stimulant as per Table 5, 0.08-0.09 weight % of jaggery, 0.1 weight % of Na2CO3, 0.08-0.09 weight % of KH2PO4 and 0.003-0.004 weight % of CaCO3 into 30-60 weight % of water is prepared. This mixture of biogas slurry, decoction and bio stimulant in water is sterilized by boiling at 120 ºC temperature for 4-5 hours.
The sterilized mixture is cooled to 30 °C temperature and the inoculum having composition as per Table 3 is added to it in an aseptic condition. Agitation and aeration are applied to this sterilized and cooled mixture for 3-5 days. Then the additive as per Table 4 is added to the mixture so obtained in weight % of 0.24-0.27.
As a diverse group of beneficial microbes aiming at enhancing soil fertility and plant nutrition, one embodiment of the present invention includes use of Azotobacter, Rhizobium, Bacillus subtilis, Bacillus megaterium, Pseudomonas fluorescence, and Mycorrhiza.
With a focus of improving nitrogen fixation and nutrient uptake by plants, another embodiment of the present invention includes use of Azospirillum, Mycorrhiza, and Rhizobium.
Aiming at addressing of multiple soil nutrient deficiencies, yet another embodiment of the present invention includes use of Azotobacter, Pseudomonas fluorescence, Bacillus subtilis, and Bacillus megaterium.
BEST METHOD OF WORKING OF THE INVENTION
Implementation of invention as per detail disclosure is discussed with the help of following examples.
Example 1
14 Kg of biogas slurry were mixed with 17.381 Kg of water and added to the container to prepare the homogenous mixture. 28 g of jaggery was dissolved in the mixture, and 35 g of Na2CO3 and 28 g of KH2PO4 were added to the mixture. 280 g of herbal mixture was added to the homogenous mixture. 1 g of CaCO3, 70 g of humic acid, 24.5 g of fulvic acid, and 17.5 g of calcified seaweed were added to the mix. The mixture was sterilized in a fermenter at 120ºC for 4-5 hours, and then the sterilized medium was cooled. Finally, 2800 g of inoculum was inoculated in the homogeneous sterilized mixture. The medium was kept for 5 days for the growth of the inoculum. Then the additives 28 g of polysorbate 20, 28 g of carboxymethyl cellulose, and 28 g of polyethylene glycol were added to the inoculated mixture. The biofertilizer so obtained is then analysed for its quality, and all the nutrients needed to grow the crops as reported in Table 9 are found in the product, which is then packed.
Example 2
20.6 Kg of biogas slurry were mixed with 10.5 Kg of water and added to the container to prepare the homogenous mixture. 31.5 g of jaggery was dissolved in the mixture, and 35 g of Na2CO3 and 31.5 g of KH2PO4 were added to the mixture. 315 g of herbal mixture was added to the homogenous mixture. 2g of CaCO3, 105 g of humic acid, 28 g of fulvic acid, and 21 g of calcified seaweed were added to the mix. The mixture was sterilized in a fermenter at 120ºC for 4-5 hours, and then the sterilized medium was cooled. Finally, 3150 g of inoculum was inoculated in the homogeneous sterilized mixture. The medium was kept for 3 days for the growth of the inoculum. Then the additives 31.5 g of polysorbate 20, 31.5 g of carboxymethyl cellulose, and 31.5 g of polyethylene glycol were added to the inoculated mixture. The biofertilizer so obtained is then analysed for its quality, and all the nutrients needed to grow the crops as reported in Table 9 are found in the product, which is then packed.
Example 3
17.412 Kg of biogas slurry were mixed with 14 Kg of water and added to the container to prepare the homogenous mixture. 28 g of jaggery was dissolved in the mixture, and 35 g of Na2CO3 and 39.75 g of KH2PO4 were added to the mixture. 295 g of herbal mixture was added to the homogenous mixture. 1.5g of CaCO3, 87.5 g of humic acid, 26.25 g of fulvic acid, and 19.25 g of calcified seaweed were added to the mix. The mixture was sterilized in a fermenter at 120ºC for 4-5 hours, and then the sterilized medium was cooled. Finally, 2975 g of inoculum was inoculated in the homogeneous sterilized mixture. The medium was kept for 4 days for the growth of the inoculum. Then the additives 29.75 g of polysorbate 20, 29.75 g of carboxymethyl cellulose, and 29.75 g of polyethylene glycol were added to the inoculated mixture. The biofertilizer so obtained is then analysed for its quality, and all the nutrients needed to grow the crops as reported in Table 9 are found in the product, which is then packed.
Table 9
Sr. No. Parameter Example 1 Example 2 Example 3
1 Color Dark brown Dark brown Dark brown
2 Foul Odor Absent Absent Absent
3 pH at 25 0C 6.8 7.0 6.8
4 C/N ratio (mg/Kg) 11 19 15
5 Moisture weight % (maximum) 96.25 96 96
6 Total organic carbon in weight % (minimum) 16 29 22
7 Total Nitrogen (as N) % 1.45 1.52 1.46
8 Total Phosphate (as P) % 0.65 1.8 1.5
9 Total Potassium (as K) % 1.0 2.5 1.98
10 Conductivity dS/m 3.0 4.0 3.5
11 Azotobacter (CFU/mL) 4.8 x 109 5.5 x 109 5.0 x 109
12 Rhizobium (CFU/mL) 8.1 x 109 9.8 x 109 7.2 x 109
13 Bacillus subtilis (CFU/mL) 6.2 x 1010 7.5 x 1010 6.9 x 1010
14 Bacillus megaterium (CFU/mL) 9.8 x 1010 1.25 x 1010 1.0 x 1010
15 Pseudomonas fluorescence (CFU/mL) 8.0 x 1010 9.0 x 1010 8.7 x 1010 , Claims:We Claim:
1. A biogas slurry based biofertilizer composition characterized in that the said composition comprising:
• 40-70% by weight of biogas slurry;
• 30-60% by weight of water;
• 0.08-0.09% by weight of Jaggery;
• 0.1% by weight of Na2CO3;
• 0.08-0.09% by weight of KH2PO4;
• 0.8-0.9% by weight of a herbal extract;
• 0.003-0.004% by weight of CaCO3;
• 8-9% by weight of an inoculum;
• 0.24-0.27% by weight of an additive and;
• 0.32-0.44% by weight of a stimulant;
Wherein
the herbal extract is a decoction containing secondary metabolite of a blend of 5.24% by weight of fenugreek, 0.17% by weight of turmeric, 0.17% by weight of asafoetida, 0.05% by weight of neem oil, 0.017% by weight of humic acid, 0.004% by weight of fulvic acid, 0.003% by weight of calcified seaweed, 0.17% by weight of cinnamon, 0.7% by weight of a micronutrient powder, 2.6% by weight of moringa powder and 3.49% by weight of aloe vera in 87.34% by weight of water,
the inoculum is consisting of 108-109 CFU/mL of Azospirillum, 108-109 CFU/mL of Rhizobium, 108-109 CFU/mL of Azotobacter, 108-109 CFU/mL of Pseudomonas fluorescens, 108-109 CFU/mL of Bacillus subtilis, 108-109 CFU/mL of Bacillus megaterium and, 106-107 spores/gram of Mycorrhizae sp,
the additive is consisting of 0.08-0.09% by weight of Polysorbate 20, 0.08-0.09% by weight of Carboxymethyl cellulose (CMC) and 0.08-0.09% by weight of Polyethylene Glycol (PEG),
the stimulant is consisting of 0.2-0.3% by weight of Humic acid, 0.07-0.08% by weight of Fulvic acid and 0.05-0.06% by weight of Calcified seaweed and,
the micronutrient powder which is the part of herbal extract is consist of 30.9% weight by weight (W/W) of Iron, 37.03% W/W of Zinc, 12.34% W/W of Copper, 12.34% W/W of Manganese, 1.23% W/W of Molybdenum and 6.2% W/W of Boron by Atomic Absorption Spectroscopy (AAS).
2. The biogas slurry based biofertilizer composition as claimed in claim 1, wherein the inoculum consisting of Rhizobium, Azotobacter, Pseudomonas fluorescence, Bacillus subtilis, Bacillus megaterium, and Mycorrhiza sp is used.
3. The biogas slurry based biofertilizer composition as claimed in claim 1, wherein the inoculum consisting of Azospirillum, Rhizobium and Mycorrhiza sp is used.
4. The biogas slurry based biofertilizer composition as claimed in claim 1, wherein the inoculum consisting of Azotobacter, Pseudomonas fluorescence, Bacillus subtilis, and Bacillus megaterium is used.
5. A method for preparation of the biogas slurry based biofertilizer composition comprising multistage fermentation process in which biogas slurry is first fermented anaerobically and then aerobically characterized in that
(a) taking the biogas slurry which is product of anaerobic fermentation,
(b) preparing a decoction containing secondary metabolite of fenugreek, turmeric, asafoetida, calcium, neem, humic acid, fulvic acid, calcified seaweed, cinnamon, micronutrient, moringa and aloe vera by boiling of a blend of 5.24% by weight of fenugreek, 0.17% by weight of turmeric, 0.17% by weight of asafoetida, 0.05% by weight of neem oil, 0.017% by weight of humic acid, 0.004% by weight of fulvic acid, 0.003% by weight of calcified seaweed, 0.17% by weight of cinnamon, 0.7% by weight of micronutrient powder, 2.6% by weight of moringa powder and 3.49% by weight of aloe vera in 87.34% by weight of water,
(c) mixing 40-70 weight % of the biogas slurry, the 0.8-0.9 weight % of decoction, 0.32-0.44 weight % a bio stimulant, 0.08-0.09 weight % of jaggery, 0.1 weight % of Na2CO3, 0.08-0.09 weight % of KH2PO4 and 0.003-0.004 weight % of CaCO3 into 30-60 weight % of water followed by sterilizing of a mixture so obtained at 120 ºC temperature for 4-5 hours,
(d) cooling down the sterilized mixture so obtained from step (c) to 30 °C temperature,
(e) adding an inoculum in the mixture so obtained from step (d) in an aseptic condition,
(f) applying agitation and aeration to a mixture so obtained from step (e) for 3-5 days and,
(g) adding 0.24-0.27 weight % of additive to a mixture so obtained from step (f).
6. The method for preparation of the biogas slurry based biofertilizer composition as claimed in claim 5, wherein the micronutrient powder which is the part of herbal extract is consist of 30.9% weight by weight (W/W) of Iron, 37.03% W/W of Zinc, 12.34% W/W of Copper, 12.34% W/W of Manganese, 1.23% W/W of Molybdenum and 6.2% W/W of Boron by Atomic Absorption Spectroscopy (AAS).