Claims:1. A highly efficient liquid organic manure composition containing animal and plant based material having C:N ratio from 15:1 to 20:1, reducing sugars, amino acids, fatty acid, organic acids derived by fermentation of sugars by naturally occurring microflora, gluconic acid, IAA – indole acetic acid and gibberellic acid, phosphates, vitamins and minerals derived from cow urine.

2. A process for the preparation of the manure composition as claimed in claim 1, which comprises fermenting by mechanical agitation without providing any external aeration source to a medium in presence of bacteria, the said medium comprising cow dung, cow urine, a carbon, nitrogen and a protein source, a long and small chain fatty acid source, the fermentation carried out at room temperature in range of 25 to 40°C, for a period of 72 to 96 hrs. Harvesting the medium, separating the liquid containing the manure produced, enriching the manure by filtration to obtain the product.

3. A process for the preparation of the manure composition as claimed in claim 2 wherein, the carbon, sulphur and vitamin source is characterised by jaggery and molasses

4. A process for the preparation of the manure composition as claimed in claim 2 wherein, the nitrogen and mineral source are paneer whey, cheese whey, and butter milk

5. A process for the preparation of the manure composition as claimed in claim 2 wherein, the protein source are gram flour, soya de-oiled cake and other proteinaceous de-oiled cakes.

6. A process for the preparation of the manure composition as claimed in claim 2 wherein, the long- and small chain fatty acids and high energy nutrition source acting as emulsifier are cow cream, cow butter, and clarified cow butter (ghee)

7. A process for the preparation of the manure composition as claimed in claim 2 wherein, the effective concentration of the liquid manure is ~3 mL per litre of diluents characterised by water
, Description:FORM 2

THE PATENTS ACT-1970
(39 of 1970)
and
The Patent Rules, 2006

COMPLETE SPECIFICATION
(See Section 10 and Rule 13)

1. TITLE OF THE INVENTION
A Highly efficient liquid organic manure composition containing
animal and plant based material and process for preparation thereof

2. APPLICANTS

a) Name BijasuAgri Res Lab LLP
b) Nationality Indian
c) Address BijasuAgri Res Lab LLP, S. No 37, Old no 33, Kondhwa Industrial estate, Behind Monginis factory, Vinu Mehta Udyan, Kondhwa, Pune-41048, Maharashtra
3. PREAMBLE TO THE DESCRIPTION
The following specification describes the nature of invention and the manner in which it is to be performed.
Title:
A Highly efficient liquid organic manure composition containing animal and plant based material and a process for preparation thereof.
Field of Invention:
The present invention relates to highly efficient organic manure composition containing animal and plant based material in the field of agriculture. More particularly, it relates to a novel organic manure composition that improves crop yield and increases organic content of soil, thereby improving its fertility. The invention also provides a process for the preparation of the said manure.
Background of the invention:
There are many chemical (synthetically derived)-free preparations that are used by farmers for agriculture purposes such as -
a) Biofertilizers - These include a single type of bacterial formulation such as nitrogen fixers, phosphate solubilizers, potassium mobilizers etc. that carry a unique trait in providing crops with the specific set of nutrients (Crook et al., 2013)
b) Organic manure- These include farm yard manure along with green manure and animal wastes, vermicompost, biogas slurry etc, which contains a complex set of organic molecules as well a bio diverse group of micro flora.
In recent years, due to increasing awareness of deteriorating soil health and environmental hazards, among urban and rural India, farmers and households are increasingly employing chemical-free 'green farming' methods.
Traditionally Cow- urine and its distillate has been used for health benefits, which has been recently scientifically interpreted in several patents such as US6410059, US6896907, US7235262, US20020164378 and US20050100610, respectively. Cow Dung is directly applied to fields or is used as a cleaning agent by smearing it on the household porch.
As a part of a green revolution farmers seek for highly efficient, ready-to-use, cost-effective formulations of organic preparations that will increase crop productivity and improve soil health. The present invention addresses all these parameters for sustainable and profitable farming and provides novel, inventive highly effective ready to use, cost effective formulations of organic preparations.
The main object of the present invention is to provide highly efficient ready to use and cost effective formulation of organic preparations in liquid form.
Another object is to provide the said formulations which increase crop productivity and improve soil health.
Still another object is to reinvent traditional methodology of farming in modern light using a combination of biotechnology and biophysical methods.
Yet another object is to maintain a high C:N ratio of 20:1 that is required to enrich soil.
Still another object is to provide composition which enriches diverse group of microbes that are naturally present in the raw materials, which constitute the major biochemical pathways to enrich soil.
Another object is to provide compositions using easily available raw materials in bulk to produce on mass-scale and cost-effective to reduce organic manure expenses of farmer and increase profitability.
Brief summary of the present invention:
Ayruvedic techniques in India have been largely restricted to human health. Ayur means life, Vedas means science, is applicable to all life forms also to Animals (Pashu) and plants (Vruksh).
The present invention focuses on plant ayruveda wherein the efficiency of traditional formulations is enhanced by modern biophysical and biotechnological processes. Cow- urine and dung are readily and abundantly available commodities but are not adequately utilized. In most cases, they are sold as and when required, while large fractions go waste resulting in biological pollution. The present invention comprises of formulation developed by fermentation of cow-based wastes with plant based carbohydrate and protein sources to increase soil health.
The present composition improves soil health is pertinent for sustainable and profitable agriculture, with results comparable to the synthetic counterparts without any hazards.
The developed organic manure composition is highly effective at low dosages requiring as little as ~3mLin 1L dilution for vegetable cash crops for family such as Solanacea and staple cereal crops. Further, it is also possible to develop the liquid organic manure specific to a particular crop by selecting specified bacteria.
The invention has also been tested with tree crops like Moringaolifera and Punicagranatum. The basis for invention is at two levels; firstly, the C: N ratio in the organic manure is maintained at 20:1 similar to the conditions suitable for a healthy soil. Secondly, soil health is directly defined by its micro biota, it is thus necessary to replenish soil microbe by enrichment and providing nutrition for growth. All these factors account in the present composition for improved crop yield and hence profitable farming.
Accordingly the present invention provides highly efficient liquid organic manure composition containing animal and plant based material having C:N ratio from 15:1 to 20:1, reducing sugars, amino acids, fatty acid, organic acids derived by fermentation of sugars by naturally occurring microflora, gluconic acid, IAA – indole acetic acid and gibberellic acid, phosphates, vitamins and minerals derived from cow urine.
The present invention also provides a process for the preparation of the manure composition which comprises fermenting by mechanical agitation without providing any external aeration source to a medium in presence of bacteria, the said medium comprising cow dung, cow urine, a carbon, nitrogen and a protein source, a long and small chain fatty acid source, the fermentation carried out at temperature of 25 to 40 °C, for a period of 72 to 96hrs., Harvesting the medium, separating the liquid containing the manure produced, enriching the manure by filtration to obtain the product.
In one of the embodiments of the present invention the carbon, sulphur and vitamin source is characterised by jiggery and molasses
In another embodiment the nitrogen and mineral source may be paneer whey, cheese whey, and butter milk
In still another embodiment the protein source may be gram flour, soya de oiled cake and other proteinaceous de-oiled cakes.
In still another embodiment the long- and small chain fatty acids and high energy nutrition source acting as emulsifier may be cow cream, cow butter, and clarified cow butter (ghee)
In another embodiment the effective concentration of the liquid manure may be ~3 mL per litre of diluents characterised by water.
The process for preparation of the liquid manure is described with reference to examples hereinbelow which are illustrative only and should not be construed to limit the scope of the present invention in any manner whatsoever.
Example 1:
200 liters organic manure is prepared by mixing the following components:
Cow Dung (50kg); Cow Urine (50liters); Paneer (cottage cheese) whey; Cream (1kg) ; Gram flour (10kg); Jaggery (10kg).

Example 2:
200 liters organic manure can also be prepared by mixing the following components-
Cow Dung (50kg); Cow Urine (40 liters); Cheese whey (50liters); Soya de-oiled cake (5kg); Jaggery (10kg).
Example 3:
200 liters organic manure can also be prepared by mixing the following components-
Cow Dung (38 kg); Cow Urine (50 liters); Cheese whey (50liters); Peanut de-oiled cake (5kg); Jaggery (10kg).
Example 4:
200 liters organic manure can also be prepared by mixing the following components-
Cow Dung (50kg); Cow Urine (50liters); Paneer (cottage cheese) whey; Cream (1kg); Mahua deoiled cake (12kg); Jaggery (10kg).
Working of the invention:
Several experiments were performed wherein a common base of cow dung, cow-urine and paneer whey was maintained as these ingredients constitute to a good amount of NPK along with a healthy C:N ratio (vide infra) of above 20:1. Furthermore, these constituents together are rich sources of beneficial bacteria and have a pH of 4.0-5.0 that is necessary for high solubility of phosphorus and micronutrients making them readily available for growth conditions.
Different carbon sources such as commercial glucose syrup, jaggery and molasses were tested against all combinations of different protein sources such as soya, rice protein and gram flour, individually.
The raw materials of the present formulation consist of organic products that are enriched by bacteria.
1. Cow dung is one of the major components of the mixture which has a C: N ratio of 20:1. This includes several digested and undigested plant based material rich in cellulose and lignin. It is also rich consortia of several gut bacteria.

2. Cow urine is another major ingredient. It is rich in nitrogenous wastes and has a C: N of 0.8:1. Furthermore, alike other excreted wastes it is rich in electrolytes.

3. Jaggery is the predominant carbon rich ingredient. It consists of up to 60% and 15% -invert and – reducing sugars, respectively. Apart from it is rich in sulphur based compounds which accounts for 2% of the content; lastly it also contains various vitamins and co-factors that are important for biomass as well as plant growth.

4. Whey is another important nitrogen rich base, which is also rich in phosphate and potassium.

5. The final ingredient is gram flour, which is an abundant protein source. It contains 22% proteins.

6. Cow cream is rich in long- and small chain fatty acids. This provides a high energy nutrition source and also acts as an emulsifier to various plant growth regulators.

Characterisation of plant growth promoting traits:

Many of these bacteria produce several useful plant-growth promoting traits. We characterized the same using NMR based metabolic profiling. Due to the complex nature of the spectrum and several peaks associated, we resorted to NMR metabolomics database BMRB to identify the metabolites based on the chemical shift resonance.

Figure: A 1D- 1H NMR based metabolic profile of the organic manure that we have formulated(red).
We identified several amino acids such as glycine, isoleucine, threonine; nucelobase such as guanine; organic acid such as acetate, pyruvate and a-keto glutarate; fatty acids such as butyric acid, decanoic acid and other organic acids such as phosphonic acid and phenylacetic acids in abundance in the NMR fingerprint. We also noticed identification of several key metabolites, like plant growth regulators comprising of indole acetate, indole pyruvate, gibberellic acids; jasmonate, salicyclate, vitamins and co-factors; nucleobases. Further investigation into the biochemical mechanism using metaboanalyst highlights the tricarboxylic acid (TCA) cycle acts as major metabolic signalling pathways to feed nitrogen containing metabolites and other organic acids.
In order to understand the role of naturally occurring microbes in the preparation, the present invention identified them by growing them on selective media and belonged to
1. Free-living motile nitrogen fixers that can fix atmospheric nitrogen and hence increase the nitrogen content of soil. These bacteria were grown on N2-free Azotobacter media. Many free living Nitrogen fixers are found in soil such as Azotobacter and Azospirillum, while others belonging to Klebsiella are also found in mammalian guts; these bacteria are also produce several different kinds of plant growth hormone and are known to stimulate plant growth.
2. Phosphate solubilizers which are sporulating (gram positive) and non-sporulating (gram negative) bacteria were cultivated on Pikosvskaya agar medium. These bacteria belong to the class of Bacillus and Pseudomonads. Solubilising phosphate under acidic conditions makes the compound readily utilizable by plants. In addition to releasing anion, the corresponding cation (mono- and or di) is also solubilised enhancing nutrient availability for plants. Additionally, these classes of bacteria can produce secondary metabolites that can inhibit pathogenic activity of various microbes thereby indirectly promoting plant growth.
3. Lactobacillus spp. on MRS agar. These classes of bacteria are fermentative bacteria found in guts of all organisms. They play a pivotal role by producing lactate, other acids and probiotic function. They have been shown to possess very strong antibacterial effect thereby killing various classes of pathogens by production of bacteriocins.
4. Anaerobic bacteria were grown on thioglycolate and clostridia media. The exact nature of these bacteria in plant growth remains unclear but the stimulatory effects may resemble conditions similar to biogas slurry, which is also rich in various organic acids, amino acids and simple carbohydrates.
Advantages of the invention:

1. It increases the fertility of the soil and hence results in increased crop yield at high dilutions of 3mLper litre. In a nutshell, it means that 1 litre of the formulation can be diluted in 330 litres of water and applied to 1 acre of field for vegetable and/or cereal crops per single application.

2. It increases the photosynthesis rate of the plant and leaves appear greener.

3. Due to the presence of immunity builders like Jasmonate and Salicyclate, the plants build resistance to microbial pathogens.