WO2013035032A2

WO2013035032A2 - Capsulated formulation with beneficial microbes for healthy plant growth

Info

Publication number: WO2013035032A2
Application number: PCT/IB2012/054552
Authority: WO
Inventors: Sharad N CHAUDHARI
Original assignee: Individual
Current assignee: Individual
Priority date: 2011-09-11
Filing date: 2012-09-04
Publication date: 2013-03-14
Anticipated expiration: 2014-03-11
Also published as: WO2013035032A3

Abstract

For healthy and timely growth of plants, along with food various factors like Nitrogen, Phosphates, Potash, Pesticides, Fungicides, Nematicides are required. Chemical based products are now being replaced by microbial based products because of serious effects on the ecology especially on plants and human beings. This invention generally relates to capsulated, water soluble consortium of microbial formulations used for activities including symbiotic & non symbiotic nitrogen fixation, phosphate solubilization, potash mobalisation and pesticidal, fungicidal as well as nematicidal. Avantages of the present invention are these microbial formulations for the use in agriculture are packed in capsules, which is user friendly form for easy handling and application, can be applied as is in capsule form or can be used as solution for spraying, wherein because of complete solubility nozzles of the sprayer does not chock, consortium of microbes in dormant stage is packed in the capsules so as to ensure compatibility, applicability at wide range of climatic conditions, have longer shelf life, can be used in any quantity without affecting quality of the remaining product in the partially used packets, fixed quantity is packed in each capsule for giving exact required dosage by farmers.

Description

Capsulated Formulation With Beneficial Microbes For Healthy Plant Growth

For healthy and timely growth of plants, along with food various factors like Nitrogen, Phosphorus, Potassium, Pesticides, Fungicides, Nematicides are required. In earlier era of agricultural revolution all these factors were given to plants using synthetic products. Though these synthetic products has played a vital role in agriculture revolution, over the years world has experienced very serious_? effects on the ecology especially on plants and human beings. This has lead to development of various biological products which can and are replacing these synthetic products. Biological products have all the advantages and do not affect ecology.

Field of Invention

This invention generally relates to capsulated, water soluble consortium of microbial formulations used for activities including but not restricted to Symbiotic and Non Symbiotic Nitrogen Fixation, Phosphate Solubilization, Potash Mobilization and Pesticidal, Fungicidal as well as Nematicidal. These formulations also include but not restricted to food generating microbes which generate nutrients for fast growth of all other microbes in these formulations. Use of these products totally eliminates use of chemical pesticides, fungicides, herbicides and the like. These products make nutrients already present in the soil or atmosphere, available to plants so that chemical fertilizers are not required to be used or required to be used in minor quantity. After application in the soil the microbes in the products grow enhancing their activity many fold. Discussion of prior art

Various products based on microbial formulations are available in the market. The major problems of these formulations are; These products are generally packed in HDPE bottles, LDPE bags, and cardboard boxes. Total contents of the whole packet is required to be used after opening of the packet. This requirement of use of total contents of the packet in one lot is a major constraint for farmers in the under developed and developing countries like India because the farm sizes generally are small and their financial spending capacity is very limited.

The shelf life of the microbial product is a major concern. In the farms products are generally stored in open space or in a covered area without any special storage conditions like air conditioning, cooling, dehumidified climate etc. Due to higher temperature (above 35°C) normally prevailing in Asian countries including India, the CFU (colony forming units) count drastically decreases in the product. The microbial activity of these formulations reduces substantially after opening the packet. In under developed and developing counties like India farm size is generally small and so, mostly total contents of the whole packet of the product is not consumed every time in one stroke.

Packaging material used to pack these products is normally permeable and allow moisture and other gases from the climate to enter the packet drastically reducing shelf life of these products.

There are chances of contamination in partially used, open packet.

During application of these products with bare hands, which is a normal procedure followed by farmers, chances of contamination are very high. In most of the cases, a single product is recommended for all crops. Generally additional food required for the timely and healthy growth of microbes in the formulation required after application is not added, affecting microbial propagation in the soil resulting in low efficiency.

These products are insoluble in water or solvents and so it is used in the suspension form which more than often chock spray nozzles as well as pores in drip irrigations system affecting its uniform and timely application.

9. Application of these products generally require further processing like dissolution etc. wherein precisely measuring recommended dosage becomes difficult for farmers in India and other developing as well as under developed countries and many times lower dosage is given affecting healthy and timely growth of plants or excess dosage is given wasting these products affecting already weak economy of the farmers.

10. Some products like bio-fungicides are packed in the form of 'Toffee or Chocolate' wherein misuse like human consumption especially by children is possible making this type of packing unsafe. Objects and Advantages of Invention

Accordingly different objects and advantages of the present invention are described below.

An object of the present invention is to provide microbial formulations packed in capsules for the use in agriculture. Other object of the present invention is to provide capsulated microbial formulations for the use in agriculture wherein microbes are in dormant stage till they come in contact with wet soil during its application. Another object of the present invention is to provide capsulated microbial formulations for the use in agriculture wherein each capsule becomes a separate tiny packing so that partially consumed open packets does not reduce microbial activity of the formulations. This feature will help small farmers in which case total contents of the packet is not required to be used in one lot.

Yet another object of the present invention is to provide capsulated microbial formulations for the use in agriculture with longer and better shelf life. Our formulations are packed in capsules protecting the contents from atmosphere so that dormant stage of the microbes in the formulations is maintained till the time of application in the field greatly enhancing shelf life of the product.

One more object of the present invention is to provide capsulated microbial formulations for the use in agriculture which do not require any specialized arrangement like temperature control, dehumidified atmosphere for its storage.

Additional object of the present invention is to provide capsulated microbial formulations for the use in agriculture wherein chances of contamination in the partially used, open packet are eliminated. More additional object of the present invention is to provide capsulated microbial formulations wherein the microbial formulation for the use in agriculture does not come in contact with human beings avoiding any chances of contamination. One more additional object of the present invention is to provide capsulated microbial formulations for the use in agriculture wherein various combinations of microbial formulations can be used for different crops. Yet one more additional object of the present invention is to provide capsulated microbial formulations for the use in agriculture which include microbes that generate food for microbes in the capsulated formulations and also for microbes present in the soil. Similarly one more additional object of the present invention is to provide capsulated microbial formulations for the use in agriculture wherein the microbes in the formulation packed in the capsule spreads and propagates to large area around the point of application. Similarly still one more additional object of the present invention is to provide capsulated microbial formulations for the use in agriculture, wherein the material of which the capsule is made, after disintegration provides organic matter useful for the growth of plants. Similarly other object of the present invention is to provide capsulated microbial formulations for the use in agriculture wherein since fixed quantity of the product is packed in the capsule exact recommended dosage can be given even by lay man working in the field without any specialized calculations or training.

The different advantages of this invention are described below: An advantage of the present invention is that these microbial formulations for the use in agriculture are packed in capsules, which is user friendly form for easy handling. Another advantage of the present invention is that this capsulated formulation for the use in agriculture can be applied as is in capsule form or can be used as solution for spraying or for drip irrigation. Yet other advantage of the present invention is to provide a capsulated microbial formulation for the use in agriculture wherein use of additional nutrients required for growth of microbes is not required.

Yet other advantage of the present invention is to provide a capsulated microbial formulation for the use in agriculture wherein the consortium of microbes in dormant stage is packed so as to ensure compatibility.

Similarly yet another advantage of the present invention is to provide a capsulated microbial formulation used in agriculture with longer shelf life.

Yet further advantage of the present invention is to provide a capsulated microbial formulation used in agriculture which can be used in any quantity without affecting quality of the remaining product in the partially used packets. Similarly more advantage of the present invention is to provide a microbial formulation used in agriculture which is packed in capsules wherein fixed quantity is packed in each capsule for giving exact required dosage by farmers in developing countries like India as well as in under developed countries. Brief Description of drawings

The invention can be understood better when read with the accompanying drawings which are described below in brief. Figure numbers from 1 to 8 (derived from Tables from 2 to 9, respectively) show graphically experimental results for microbial propagation in terms of colony count versus time in hours for PHH-based formulations. Figure numbers from 9 to 16 (derived from Tables from 10 to 17, respectively) show graphically experimental results for microbial propagation in terms of colony count versus time in hours for Talcum - based formulations.

Figure numbers from 17 to 24 (derived from Tables from 18 to 25, respectively) show graphically experimental results for microbial propagation in terms of colony count versus time in hours for Lignite - based formulations.

Figure numbers from 25 to 32 (derived from Tables from 26 to 33, respectively) show graphically experimental results for microbial propagation in terms of colony count versus time in hours for Water-based formulations.

Figure numbers from 33 to 40 (derived from Tables from 34 to 41 , respectively) show graphically experimental results for shelf- life study for 10 months duration for PHH-based formulation.

Figure number 41 (derived from Table 42) show graphically microbial propagation of Encounter, in soil at different distances from spot of application.

Figure numbers from 42 to 49 (derived from Tables from 43 to 50, respectively) show graphically comparative study of each formulation with four different bases. Summary of the invention

For healthy and timely growth of plants, along with food various factors like Nitrogen, Phosphorus, Potassium, Pesticides, Fungicides, Nematicides are required. Chemical based products are now being replaced by microbial based products because of serious effects on the ecology especially on plants and human beings. This invention generally relates to capsulated, water soluble consortium of microbial formulations used for activities including symbiotic & non symbiotic nitrogen fixation, phosphate solubilization, potash mobilization and pesticidal, fungicidal as well as nematicidal. Avantages of the present invention are these microbial formulations for the use in agriculture are packed in capsules, which is user friendly form for easy handling and application, can be applied as is in capsule form or can be used as solution for spraying, wherein because of complete solubility nozzles of the sprayer does not chock, consortium of microbes in dormant stage is packed in the capsules so as to ensure compatibility, applicability at wide range of climatic conditions, have longer shelf life, can be used in any quantity without affecting quality of the remaining product in the partially used packets, fixed quantity is packed in each capsule for giving exact required dosage by farmers.

Detailed description of the invention

The present application discloses mix of products including but not restricted to following products used for various applications packed in capsules of various types including but not restricted to vegetable capsules, gelatin capsules, soft capsules, gel capsules etc. of various sizes and shapes including but not restricted to 000, 00, 0, 1, 2, 3, 4 etc. These products are defined below.

Definitions

The following definitions shall apply to the description given in this specification. Nitroset is defined as a biologically derived first bio-fertilizer, capably effecting nitrogen fixation, packaged in a first capsule (CP1).

Phossol is defined as a biologically derived second bio-fertilizer, capably effecting phosphate solubilization, packaged in a second capsule (CP2).

Potmob is defined as a biologically derived third bio -fertilizer, capably effecting potash mobilization, packaged in a third capsule (CP3). Encounter is defined as a biologically derived bio-pesticide, capably effecting crop- protection, packaged in a fourth capsule (CP4).

Rotuccider is defined as a biologically derived bio-fungicide, capably effecting crop- protection, packaged in a fifth capsule (CP5).

Nemuccider is defined as a biologically derived bio-nemuccider, capably effecting crop-protection, packaged in a sixth capsule (CP6).

Encounter-100 is defined as a biologically derived bio-foliar protector, capably effecting foliar - protection, packaged in a seventh capsule (CP7).

Bravo is defined as a biologically derived bio-nutrition, capably effecting nutrition for microbes and crops as well, packaged in an eight capsule (CP8). A water soluble-compound PHH is defined as a base or a carrier, comprising (2R, 3S, 4R, 5R)-2, 3, 4, 5, 6 - Pentahydroxyhexanal to be impregnated with relevant microbes. SMS is defined as sodium metasilicate.

More details of these defined products are given below:

(1) Nitroset: Consortia of symbiotic and non symbiotic nitrogen fixing microbes including but not limited to Azotobacter chroococcum, Azotobacter vinelindii, Azotospirillum lipoferum, Rhizobium, Acetobacter aceti, Azospirillium spp. as a single entity or combination thereof in equal proportion, impregnated on a carrier which is water soluble selected from the list of carriers including, but not limited to PHH, a-(lR,2R,3S,4S,5R)-glucopyranosyl- -(2S,3S,4S,5R)- fructofuranoside, D-arabino-hexulose, stevia sugar, etc. which converts this mixture into water soluble powder which is packed in capsules of various types including but not restricted to vegetable capsules, gelatin capsules, soft capsules, gel capsules etc. of various sizes and shapes including but not restricted to 000, 00, 0, 1, 2, 3, 4 etc. These microbes are used to fix nitrogen from the atmosphere and in the soil so that plants can use the nitrogen required for their healthy and timely growth. This formulation can be used as nitrogen fixers for the plants including but not limited to monocotyledonous, dicotyledonous, sugar and submerged plants. The crops that can be benefited include but not limited to horticulture, floriculture, commercial crops, spices and condiments, vegetables, cereal, pulses and all other crops.

(2) Phossol: Consortia of phosphate solubilizing microbes including but not limited to Pseudomonas putida, Bacillus megaterium as a single entity or combination thereof in equal proportion, impregnated on a carrier which is water soluble selected from the list of carriers including but not limited to PHH, a-(lR,2R,3S,4S,5R)-glucopyranosyl- -(2S,3S,4S,5R)-fructofuranoside, D-arabino-hexulose, stevia sugar, etc. which converts this mixture into water soluble powder which is packed in capsules of various types including but not restricted to vegetable capsules, gelatin capsules, soft capsules, gel capsules etc. of various sizes and shapes including but not restricted to 000, 00, 0, 1, 2, 3, 4 etc. These microbes are used to convert insoluble phosphates in to soluble phosphates in the soil so that plants can use the phosphorus required for their healthy and timely growth. These bacteria can be used as phosphate solubilizes for the plants. The crops that can be benefited include but not limited to horticulture, floriculture, commercial crops, spices and condiments, vegetables, cereal and pulses and all other crops.

(3) Potmob: Potash mobilizing microbes including but not limited to Frateuria Aurentia as a single entity, impregnated on a carrier which is water soluble selected from the list of carriers including but not limited to PHH, a- (lR,2R,3S,4S,5R)-glucopyranosyl- -(2S,3S,4S,5R)-fructofuranoside, D- arabino-hexulose, stevia sugar, etc. which converts this mixture into water soluble powder which is packed in capsules of various types including but not restricted to vegetable capsules, gelatin capsules, soft capsules, gel capsules etc. of various sizes and shapes including but not restricted to 000, 00, 0, 1, 2, 3, 4 etc. These microbes are used to mobilize potash so that plants can use the potash in the soil required for their healthy and timely growth. This formulation can be used as potash mobilisers for the plants. The crops that can be benefited include but not limited to horticulture, floriculture, commercial crops, spices and condiments, vegetables, cereal and pulses and all other crops.

(4) Encounter: Microbes acting as bio-pesticides including but not limited to Pseudomonas fluorescence as a single entity, impregnated on a carrier which is water soluble selected from the list of carriers including but not limited to PHH, a-(lR,2R,3S,4S,5R)-glucopyranosyl- -(2S,3S,4S,5R)-fructofuranoside, D-arabino-hexulose, stevia sugar, etc. which converts this mixture into water soluble powder which is packed in capsules of various types including but not restricted to vegetable capsules, gelatin capsules, soft capsules, gel capsules etc. of various sizes and shapes including but not restricted to 000, 00, 0, 1, 2, 3, 4 etc. These bacteria have strong pesticidal activity and are used to control root and wilt diseases and to expel and develop essential amino acid IIA, gibralic acid with some cytokinines that helps dissolve binded phophates in the soil. These microbes can be used as bio-pesticides for the plants. The crops that can be benefited include but not limited to horticulture, floriculture, commercial crops, spices and condiments, vegetables, cereal and pulses and all other crops.

(5) Encounter- 100: Consortia of microbes acting as bio -pesticides including but not limited to Metarhizium anisopliae, Beauveria bassiana, Verticillium lecanli as a single entity or combination thereof in equal proportion, impregnated on a carrier which is water soluble selected from the list of carriers including but not limited to PHH, a-(lR,2R,3S,4S,5R)- glucopyranosyl- -(2S,3S,4S,5R)-fructofuranoside, D-arabino-hexulose, stevia sugar, etc. which converts this mixture into water soluble powder which is packed in capsules of various types including but not restricted to vegetable capsules, gelatin capsules, soft capsules, gel capsules etc. of various sizes and shapes including but not restricted to 000, 00, 0, 1, 2, 3, 4 etc. These microbes act against termites, Witchetty grub, Milibug, Thrips, Sucking insects, all Borers, Scraping insects, Beetles, White ants, Scales, Aphids etc. and destroy. These microbes can be used as bio-pesticides for the plants. The crops that can be benefited include but not limited to horticulture, floriculture, commercial crops, spices and condiments, vegetables, cereal and pulses and all other crops.

(6) Rottucider: Consortia of microbes acting as bio-fungicides including but not limited to Tricoderma Viridae or Trichoderma Hurzianum as a single entity, impregnated on a carrier which is water soluble selected from the list of carriers including but not limited to PHH, a-(lR,2R,3S,4S,5R)- glucopyranosyl- -(2S,3S,4S,5R)-fructofuranoside, D-arabino-hexulose, stevia sugar, etc. which converts this mixture into water soluble powder which is packed in capsules of various types including but not restricted to vegetable capsules, gelatin capsules, soft capsules, gel capsules etc. of various sizes and shapes including but not restricted to 000, 00, 0, 1, 2, 3, 4 etc. These microbes have strong fungicidal activity and are used to fight and kill fungi with its unique mode of action called Mycoparasitism, Antibiosis and competition. These bacteria can be used as bio-fungicides for the plants. The crops that can be benefited include but not limited to horticulture, floriculture, commercial crops, spices and condiments, vegetables, cereal and pulses and all other crops.

(7) Nemuccider: Capsulated consortia of microbes acting as bio-nematicides including but not limited to Paecilomyces Lilacinus, Arthrobotrys spp as a single entity or combination thereof in equal proportion, impregnated on a carrier which is water soluble selected from the list of carriers including but not limited to PHH, a-(lR,2R,3S,4S,5R)-glucopyranosyl- -(2S,3S,4S,5R)-fructofuranoside, D- arabino-hexulose, stevia sugar, etc. which converts this mixture into water soluble powder which is packed in capsules of various types including but not restricted to vegetable capsules, gelatin capsules, soft capsules, gel capsules etc. of various sizes and shapes including but not restricted to 000, 00, 0, 1, 2, 3, 4 etc. These microbes have strong nematicidal activity and are used to fight and kill nematode parasites in the plants. These microbes can be used as bio- nematicides for the plants. The crops that can be benefited include but not limited to horticulture, floriculture, commercial crops, spices and condiments, vegetables, cereal and pulses and all other crops. (8) Bravo: Consortia of microbes acting as food generators including but not limited to Bacillus subtilis, Bacillus polymyxa, Pseudomonas fluorescens, Pseudomonas putida, as a single entity or combination thereof in equal proportion, impregnated on a carrier which is water soluble selected from the list of carriers including but not limited to PHH, a-(lR,2R,3S,4S,5R)- glucopyranosyl- -(2S,3S,4S,5R)-iructofuranoside, D-arabino-hexulose, stevia sugar, etc. which converts this mixture into water soluble powder which is packed in capsules of various types including but not restricted to vegetable capsules, gelatin capsules, soft capsules, gel capsules etc. of various sizes and shapes including but not restricted to 000, 00, 0, 1, 2, 3, 4 etc. These microbes generate nutrients required for the healthy and timely growth of all the microbes in the formulations included in all the products, during and after application. These microbes can be used as bio-food generators for the plants as well. The inventor has developed a package/ kit including all the above formulations of the different useful microbes packed in capsules of various types including but not restricted to vegetable capsules, gelatin capsules, soft capsules, gel capsules etc. of various sizes and shapes including but not restricted to 000, 00, 0, 1, 2, 3, 4 etc. which serves as fertilizers, as protecting agents as well as food for microbes in formulations as well for plants. The package includes the following:

Table 1: Composition of present invention

Details are;

A. Fertiliser set in the kit

a. Nitrogen fixers: Nitroset

i. Symbiotic N-fixer: Azotospirillum lipoferum (useful for monocotyledonous crops), Rhizobium (useful for dicotyledonous crops, Acetobacter aceti (useful for sugar crops)

ii. Nonsymbiotic N-fixer: Azotobacter chroococcum

(useful for monocotyledonous crops), Azotobacter vinelindii (useful for monocotyledonous crops), Azospirillium spp (useful for submerged crops) b. Phosphate solubilises: Phossol

i. Pseudomonas putida (Phosphate solubilising microbes) ii. A. niger or B. megaterium

c. Potash mobilisers: Potmob- i. Frateuria aurentina

Protection set in the kit

a. Encounter

i. Pseudomonas fluorescence

b. Rotuccider

ii. Trichoderma Viridae or Trichoderma Hurzianum

c. Nemuccider

i. Pacilomyces liliacinus

ii. Bacillus subtilis

d. Foliar protectors: Encounter- 100

Hi. Beauveria bassiana

iv. Metarhizium anisoplae v. Verticillium anosoplae

C. Bio-Nutritional food for microbes and plants: Bravo

i. Bacillus subtilis

ii. Bacillus polymyxa

Hi. Pseudomonas fluorescens

iv. Pseudomonas putida

1. As compared to solo application of the beneficial microbe the consortia is always proven to be better in performance. Our kit is best example of microbial consortia for specific crop. Putting all microbes in single formulation may result in antagonism. That's why we are putting different microbes separately in separate capsules.

2. The use of capsule formulation proves to have more shelf life than traditional products.

3. Due to provision of microbial selective food, the performance of the microbes found to be improved.

4. The capsule form is found to be easy for handling /application in field

5. The capsule provides additional carbon source to the microbes, which resulted in improvement in CFU count in soil.

6. Due to capsule the contamination is found to be very low.

7. The capsule delivered the active ingredient slowly in soil as compare to traditional formulations.

8. The product inside capsule is soluble powder, which is readily soluble in water. It prevents nozzle choking.

The inventor has carried out extensive experimentation. The results of this very large number of experiments are presented in the following 52 tables and 49 figures. Results obtained for all PHH based formulations are as follows;

Table 2: Microbial Propagation Results of Nitroset; PHH based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 1 by arrow)

Table 3: Microbial Propagation Results of Phossol; PHH based

Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 2 by arrow)

Table 4: Microbial Propagation Results of Potmob; PHH based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 3 by arrow)

Table 5: Microbial Propagation Results of Encounter; PHH based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 4 by arrow) Table 6: Microbial Propagation Results of Rotuccider; PHH based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 5 by arrow) Table 7: Microbial Propagation Results of Nemuccider; PHH based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 6 by arrow) Table 8: Microbial Propagation Results of Encounter-100; PHH based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 7 by arrow) Table 9: Microbial Propagation Results of Bravo; PHH based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 8 by arrow) Results obtained for all talcum based formulations are as follows;

Table 10: Microbial Propagation Results of Nitroset; Talcum based

* Colonies are dense and are beyond quantification; Density increases many fold (This phenomenon is represented in the figure 9 by arrow)

Table 11: Microbial Propagation Results of Phossol; Talcum based

* Colonies are dense and are beyond quantification; * Density increases many fold (This phenomenon is represented in the figure 10 by arrow) Table 12: Microbial Propagation Results of Potmob; Talcum based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 1 1 by arrow)

Table 13: Microbial Propagation Results of Encounter; Talcum based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 12 by arrow) Table 14: Microbial Propagation Results of Rotuccider; Talcum based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 13 by arrow)

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 14 by arrow) Table 16: Microbial Propagation Results of Encounter- 100; Talcum based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 15 by arrow)

Table 17: Microbial Propagation Results of Bravo; alcum based

* Colonies are dense and are beyond quantification; ^'■ Density increases many fold (This phenomenon is represented in the figure 16 by arrow) Results obtained for all lignite based formulations are as follows;

Table 18 Microbial Propagation Results of Nitroset; Lignite based

* Colonies are dense and are beyond quantification; ** Density increases many fold

(This phenomenon is represented in the figure 17 by arrow)

Table 19: Microbial Propagation Results of Phossol; Lignite based

(This phenomenon is represented in the figure 18 by arrow) Table 20: Microbial Propagation Results of Potmob; Lignite based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 19 by arrow)

Table 21: Microbial Propagation Results of Encounter; Lignite based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 20 by arrow) Table 22: Microbial Propagation Results of Rotuccider; Lignite based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 21 by arrow) Table 23: Microbial Propagation Results of Nemuccider; Lignite based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 22 by arrow) Table 24: Microbial Propagation Results of Encounter- 100; Lignite based

(This phenomenon is represented in the figure 23 by arrow)

Table 25: Microbial Propagation Results of Bravo; Lignite based

(This phenomenon is represented in the figure 24 by arrow) Results obtained for all water based formulations are as follows;

Table 26: Microbial Propagation Results of Nitroset; Water based

(This phenomenon is represented in the figure 25 by arrow)

Table 27: Microbial Propagation Results of Phossol; Water based

(This phenomenon is represented in the figure 26 by arrow) Table 28: Microbial Propagation Results of Potmob; Water based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 27 by arrow)

Table 29: Microbial Pro a ation Results of Encounter Water based

* Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 28 by arrow) Table 30: Microbial Propagation Results of Rotuccider; Water based

* Colonies are dense and are beyond quantification; Density increases many fold

(This phenomenon is represented in the figure 29 by arrow)

Table 31: Microbial Pro a ation Results of Nemuccider Water based

*Colonies are dense and are beyond quantification; ** Density increases many fold (This phenomenon is represented in the figure 30 by arrow) Table 32: Microbial Propagation Results of Encounter- 100; Water based

(This phenomenon is represented in the figure 31 by arrow) Table 33: Microbial Propagation Results of Bravo; Water based

(This phenomenon is represented in the figure 32 by arrow) Results of shelf life study of all of our formulations (PHH based) are as follows; Table 34: Shelf life study ofNitroset (PHH based) (Reference figure 33)

Colony count is converted into plot able figures (power of 10 is converted into decimal point) Table 35: Shelf life study of Phossol (PHH based) (Reference figure 34)

Colony count is converted into plot able figures

Table 36: Shelf life study of Potmob (PHH based) (Reference figure 35)

Colony count is converted into plot able figures Table 37: Shelf life study of Encounter (PHH based) (Reference figure 36)

Colony count is converted into plot able figures

Table 38: Shelf life study of Rotuccider (PHH based) (Reference figure 37)

Colony count is converted into plot able figures

Table 39: Shelf life study of Nimmucider (PHH based) (Reference figure 38)

Colony count is converted into plot able figures Table 40: Shelf life study of Encounter- 100 (PHH based) (Reference figure 39)

* Colony count is converted into plot able figures

Table 41: Shelf life study of Bravo (PHH based) (Reference figure 40)

* Colony count is converted into plot able figures

Results of microbial propagation of our capsulated formulation at any given point away from point of application for Encounter is tabulated as under,

Table 42: Comparison: Microbial propagation of various formulations of Encounter soil at particular distance from spot of application (Reference figure 41)

Following tables show comparison of our PHH based products with our talcum based, lignite based and water based products. Commercial formulations are based on talcum, lignite and water. Table 43: Nitroset Colony count X 10^"7 (Ref Table no. 2, 10, 18, 26; figure no 1, 9, 17, 25)

(This phenomenon is represented in the figure 42 by arrow)

Table 44: Phossol Colony count X IP^"7 (Ref Table no 3, 11, 19, 27; figure no 2, 10, 18, 26)

* Colonies are dense and are beyond quantification; Density increases many fold

(This phenomenon is represented in the figure 43 by arrow)

Table 45: Potmob Colony count X 10^"7 (Ref Table no 4, 12, 20, 28; figure no 3, 11, 19, 27)

* Colonies are dense and are beyond quantification; Density increases many fold

(This phenomenon is represented in the figure 44 by arrow) Table 46: Encounter Colony count X 10⁷(RefTable no 5, 13, 21, 29; figure no 4, 12, 20, 28)

(This phenomenon is represented in the figure 45 by arrow)

Table 47: Rotuccider Colony count X 10⁷(RefTable no 6, 14, 22, 30; figure no 5, 13, 21, 29)

Colonies are dense and are beyond quantification; ** Density increases many fold

(This phenomenon is represented in the figure 46 by arrow) Table 48; Nemuccider Colony count X 10⁷(Ref table no 7, 15, 23, 31; figure no 6, 14, 22, 30)

* Colonies are dense and are beyond quantification; Density increases many fold

(This phenomenon is represented in the figure 48 by arrow) Table 50;Bravo Colony Count X IP^"7 (Ref Table no 9, 17,25,33; figure no 8, 16, 24, 32)

(This phenomenon is represented in the figure 49 by arrow) Following table shows comparison of our capsulated PHH based formulations with our formulations based on talcum, lignite and water, which are used in available products.

Table 51:

Distinguishing features of this invention

The distinguishing features of the present invention are clearly brought out in the following description, along with references to the extensive experimental results obtained by the inventor, as shown in respective figures and given in the tabular presentations in this specification. The details presented below clearly bring out the much higher effectiveness of the capsulated formulations of the present invention as compared with commercially available present formulations of the prior art, using different prior art bases such as talcum, lignite and water as carrier for microbes. A. Following distinguishing features can be clearly observed from the table 51:

1. The Nitroset of present invention formulation (PHH-based) propagated 40.3% more (Ref. table 2; figure 1) than Talcum based (prior art) formulation (Ref. table 10; figure 9); 41.2% more than Lignite based (prior art) formulation (Ref. table 18; figure 17) and 56.3% more than Water based (prior art) (Ref. table 26; figure 25) formulation.

The Phossol of present invention formulation (PHH-based) propagated 41.8% more (Ref. table 3; figure 2) than Talcum based (prior art) formulation (Ref. table 1 1 ; figure 10); 26.5% more than Lignite based (prior art) formulation (Ref. table 19; figure 18) and 49.0% more than Water based (prior art) (Ref. table 27; figure 26) formulation.

The Potmob of present invention formulation (PHH-based) propagated 37.4% more (Ref. table 4; figure 3) than Talcum based (prior art) formulation (Ref. table 12; figure 1 1); 33.6% more than Lignite based (prior art) formulation (Ref. table 20; figure 19) and 47.7% more than Water based (prior art) (Ref. table 28; figure 27) formulation. The Encounter of present invention formulation (PHH - based), propagated 36.8% more (Ref. table 5; figure 4) than Talcum based (prior art) formulation (Ref. table 13; figure 12); 22.8% more than Lignite based (prior art) formulation (Ref. table 21 ; figure 20) and 43.9% more than Water based (prior art) (Ref. table 29; figure 28) formulation.

The Rotuccider of present invention formulation (PHH-based) propagated 44.4% more (Ref. table 6; figure 5) than Talcum based (prior art) formulation (Ref. table 14; figure 13); 45.3% more than Lignite based (prior art) formulation (Ref. table 22; figure 21) and 56.4% more than Water based (prior art) (Ref. table 30; figure 29) formulation.

The Nemuccider of present invention formulation (PHH-based) propagated 47.0% more (Ref. table 7; figure 6) than Talcum based (prior art) formulation (Ref. table 15; figure 14); 47.0% more than Lignite based (prior art) formulation (Ref. table 23; figure 22) and 54.8% more than Water based (prior art) (Ref. table 31 ; figure 30) formulation.

The Encounter- 100 of present invention formulation (PHH-based) propagated 42.6% more (Ref. table 8; figure 7) than Talcum based (prior art) formulation (Ref. table 16; figure 15); 41.7% more than Lignite based (prior art) formulation (Ref. table 24; figure 23) and 56.5% more than Water based (prior art) (Ref. table 32; figure 31) formulation.

The Bravo of present invention formulation (PHH-based) propagated 59.6% more (Ref. table 9; figure 8) than Talcum based (prior art) formulation (Ref. table 17; figure 16); 42.2% more than Lignite based (prior art) formulation (Ref. table 25; figure 24) and 51.4% more than Water based (prior-art) (Ref. table 33; figure 32) formulation.

These results clearly and unambiguously show that the capsulated formulations which are based on PHH propagates much more than Talcum based, Lignite based and Water based formulations under comparable experimental conditions.

Field trials were conducted for our bio -fertilizers Nitroset, Psossol, Potmob, bio- nutrient Bravo and bio-fungicides Rotuccider and bio- nematicider Nemuccider on wheat. Results are tabulated as under and are self-explanatory.

Table 52:

* In the case of control even though the length is comparable with other group of plants, the deviation from the average is large while in others deviation from the average is small. Thus growth of roots in other groups of plants is uniform while that in the control has lot of variations.

B. Following distinguishing features can be clearly observed from the experimental results presented in the table 52:

(A) In case of the Nitroset, Phossol, Potmob and Bravo of present invention formulations (PHH-based);

1. % germination is 98% as against 40% in control Avearage height of plants is 240mm in comparison to 170mm in contriol

% green leaves is 99.5% as against 33.3% in the control

Number of leaves are 69.4% over the control

Average length of roots are 75.4mm as against 60mm in the control

Number of roots are 245 as against 60 in control

In case of the Rotuccider of present invention formulation (PHH-based);

1. % germination is 70% as against 40% in control

2. Avearage height of plants is 220mm in comparison to 170mm in contril

3. % green leaves is 98.1% as against 83.3% in the control

4. Number of leaves are 33.3% over the control

5. Average length of roots are 60mm as against 60mm in the control

6. Number of roots are 140 as against 60 in control (C) In case of the Nemuccider of invnetion formulation (PHH-based);

1. % germination is 96% as against 40% in control

2. Avearage height of plants is 240mm in comparison to 170mm in control

3. % green leaves is 99.5% as against 83.3% in the control

4. Number of leaves are 68.7% over the control

5. Average length of roots are 50mm as against 60mm in the control

6. Number of roots are 144 as against 60 in control

These results clearly and unambiguously show that the capsulated formulations of the presnt invention, which are based on PHH propagates very much more than Talcum based, Lignite based and Water based formulations under comparable experimental conditions. C. Following distinguishing features can be easily observed from the table 42; figure 41:

These experimental data clearly indicate that the capsulated formulation of Encounter of present invention formulation (PHH-based) propagates better than other formulations; in particular 38% better than Talcum based formulation, 40% better than Lignite based and 28% better than Water based formulation at distance of 14cm. from the spot of application.

D. Following distinguishing features can be clearly observed from the respective tables (table 34 to 41; figure 33 to 40):

All of present invention formulations (PHH-based), that is, Nitroset, Phossol, Potmob, Encounter, Rotuccider, Nemuccider, Encouner-100 and Bravo are stabilised even after 7 to 8 months from month of preparation, thereby indicating very long shelf life.

Analysis of the soil in which all the above testing is done is as under.

1. pH: 7.4

2. Hardness: 1.94 m mhos/cm

3. Water holding capacity: 54.51%

4. Density: 1.33

5. Available Nitrogen: 159.94Kg/Hectare

6. Organic Carbon: 0.24%

7. Available Phosphorous: 36.47Kg/Hectare

8. Available Potassium: 450.78Kg/Hectare

9. Available Calcium: 107.1mg/100g

10. Available Magnesium: 128.27mg/100g

1 1. Available Sodium: 3.05mg/100g

12. Available Lime: 2.4mg/100g

13. Copper: 0.53ppm 14. Iron: 1.61ppm

15. Manganese: 2.35ppm

16. Zinc: 0.19ppm

According to the present invention, is provided a process for preparation of microbial formulations and encapsulation thereof comprising the following steps that is preparation of formulations, encapsulation of the formulations.

Steps of preparation of Nitroset, Phossol, Potmob, Encounter and Bravo:

1. Preparing 'Nutrient Broth' for bacteria (medium);

2. charging distilled water in to conical flasks;

3. charging 'Nutrient Broth' in to conical flasks at predetermined first temperature;

4. autoclaving flasks at predetermined second temperature for predetermined first duration;

5. inoculating each strain separately under aseptic conditions at predetermined third temperature separately in different flasks;

6. transferring each of said flasks of step 5 separately to a shaker and maintaining each of the transferred flasks on said shaker at a predetermined fourth temperature for a predetermined second duration;

7. transferring flasks to incubator;

8. incubating at predetermined fifth temperature for predetermined third duration;

9. adding SMS solution of predetermined first concentration by weight of SMS, per weight of broth;

10. mixing each broth with PHH powder to get predetermined second concentration of broth per the weight of the mixture with a predetermined first potency;

11. drying at predetermined sixth temperature for predetermined fourth duration; 12. mixing of dried products;

13. packing predetermined quantity of the mixed and dried products into capsules of predetermined size, of predetermined shape and made up of predetermined material.

In the above mentioned 'Preparation of Nitroset; Nitrogen fixers, Symbiotic N- fixer and Nonsymbiotic N-fixer- Azotobacter chroococcum, Azotobacter vinelindii, Azotospirillum lipoferum, Rhizobium, Acetobacter aceti, Azospirillium spp., Phossol; Phosphate solubilises, Pseudomonas putida (Phosphate solubilising microbes) and A. niger or B. megaterium; Potmob; Potash mobilisers, Frateuria aurentin, Encounter; Crop protectors, Pseudomonas fluorescence and Bravo; Nutritional food for microbes and plants, Bacillus subtilis, Bacillus polymyxa, Pseudomonas fluorescens and Pseudomonas putida' step, the parameters which are used, are given later on the following pages. Steps of preparation of Rotuccider, Nemuccider and Encounter-100:

1. Preparing 'Potato Dextrose Broth' for fungus (medium);

2. charging distilled water in to conical flasks;

3. charging 'Potato Dextrose Broth' in to conical flasks at a predetermined first temperature;

4. autoclaving flasks at a predetermined second temperature for predetermined first duration;

5. inoculating each strain separately under aseptic conditions at a predetermined third temperature separately in different flasks;

6. transferring each of said flasks of step 5, separately to a shaker and maintaining each of the transferred flasks on said shaker at a predetermined seventh temperature for a predetermined fifth duration;

7. transferring flasks to an incubator; 8. incubating at a predetermined fifth temperature for a predetermined sixth duration;

9. adding SMS solution of a predetermined first concentration by weight of SMS per weight of said broth;

10. mixing said broth with PHH powder to get a predetermined second concentration of broth by weight, per weight of the mixture with a predetermined first potency;

11. drying said mixture of step 10 at a predetermined sixth temperature for a predetermined fourth duration;

12. mixing of dried products;

13. packing predetermined quantity of each of the dried products into separate capsules of predetermined size, of predetermined shape and made up of predetermined material. In the above mentioned 'Preparation of Rotuccider; Crop protectors, Trichoderma Viridae or Trichoderma Hurzianum, Nemuccider; Crop protectors, Pacilomyces liliacinus, Bacillus subtilis and Encounter-100; Foliar protectors, Beauveria bassiana, Metarhizium anisoplae and Verticillium anosoplae'' step, the following parameters are used. As will be apparent to a person skilled in the art, the process of the invention can proceed in a very broad range of parameters.

A range of said first temperature is from 18°C to 37°C, preferably from 20°C to 32°C, and more preferably from 25°C to 30°C. A range of said second temperature is from 74°C to 125°C, preferably from 94°C to 124°C and more preferably from 115°C to 121°C. A range of said third or fifth or seventh temperature is from 10°C to 35°C, preferably from 15°C to 32°C more preferably from 25°C to 30°C.

A range of said fourth temperature is from 10°C to 35°C, preferably from 15°C to 32°C, more preferably from 25°C to 30°C.

A range of said sixth temperature is from 20°C to 37°C, preferably from 25°C to 35°C, more preferably between 27°C to 32°C. A range of said first duration is from 20 minutes to 90 minutes preferably from 25 minutes to 60 minutes and more preferably from 30 minutes to 45 minutes.

A range of said second or third duration is from 1 day to 7 days, preferably from 2 days to 6 days, more preferably from 3 days to 4 days.

A range of said fourth duration is from 5 hours to 24 hours, preferably from 6 hours to 18 hours and more preferably from 7 hours to 10 hours

A range of said fifth or sixth duration is from 3 days to 10 days preferably from 4 days to 8 days, and more preferably from 5 days to 7 days.

A range of said first concentration of SMS by weight is from 1% to 5%, preferably from 2% to 4% and more preferably from 2.5% to 3.5% of weight of broth.

A range of said second concentration of broth by weight is from 5% to 10%, preferably from 6% to 9% more preferably from 7% to 8% of weight of mixture of broth and PHH. A range of said first potency is from 1 x 10^~7 to 5 x 10^~7, preferably from

2 x 10^~7 to 4 x 10^~7, and more preferably from 2 x 10^~7 to 3 x 10^~7.

A range of said predetermined required quantity of said dried material is from 0.1 gm to 2 gm, preferably from 0.5 gm to 1.5 gm, more preferably from 0.8 gm to 1.2 gm.

A range of said predetermined size is selected from the group consisting of 000, 00, 0, 1, 2, 3, 4 and such other sizes. A range of said predetermined shapes is selected from the group consisting of appropriate shapes.

A range of said predetermined material is selected from the group consisting of vegetable capsules, gelatin capsules, soft capsules, gel capsules and such other capsules.

A number of illustrative and non-limitative embodiments of the invention will now be described, with reference to the examples below.

Example 01

Distilled water 300ml and 'Nutrient Broth' 3.9g is charged in each of 24 conical flaks of Hit. capacity. All flasks are autoclaved at 121°C, 15psi for 15min. and cooled to room temperature. At room temperature 'Bacterial Broths' of various strains are inoculated separately in aseptic condition in equal number of flasks based on number of strains mixed, and all the flasks are transferred to shaker and kept on shaker for 3 days. All flasks are then transferred to incubator and maintained at 25°C to 30°C for 2 days. After completion of incubation period each flask is added 140g of 2% SMS. Broths are then mixed separately with equal proportion out of total lOOKg PHH powder, dried for 7 to 8 hrs. and mixed. Example 02

Distilled water 300ml, 'Potato Dextrose Broth' 7.2g and Twin 80; 0.05% is charged in each of 24 conical flaks of Hit. capacity. All flasks are autoclaved at 121 °C for 1 hr. and cooled to room temperature. At room temperature 'Fungal Broths' of various strains are inoculated separately in aseptic condition in equal number of flasks based on number of strains mixed, and all the flasks are transferred to shaker and kept on shaker for 3 days. All flasks are then transferred to incubator and maintained at 25°C to 30°C for 2 days. After completion of incubation period each flask is added 140g of 2% SMS. Broths are then mixed separately with equal proportion out of total lOOKg PHH powder, dried for 7 to 8 hrs. and mixed.

Example 03

Four trays of same dimensions with holes for draining excess water are filled with soil from farm. In the 1^st tray 50 wheat seeds along with Nitroset, Phossol, Potmob, Bravo were sowed, in the 2^nd tray 50 wheat seeds along with Rotucider were sowed, in the 3^rd tray 50 wheat seeds along with Nemuccider were sowed and in the 4^th tray 50 wheat seeds without any of the products were sowed which is used as a control. All the trays were regularly and in same way irrigated. After 15 days all the measurements of the plants were recorded.

Although the invention has been described with reference to certain preferred embodiments, the invention is not meant to be limited to those preferred embodiments. Alterations to the preferred embodiments described are possible without departing from the spirit of the invention. However, the process and composition described above are intended to be illustrative only, and the novel characteristics of the invention may be incorporated in other forms without departing from the scope of the invention.

Claims

1) A capsulated formulation with beneficial microbes for healthy plant growth, comprising: a. a biologically derived first bio -fertilizer capably effecting nitrogen fixation, packaged in a first capsule (CP1), b. a biologically derived second bio -fertilizer capably effecting phosphate solubilisation packaged in a second capsule (CP2), c. a biologically derived third bio-fertilizer capably effecting potash mobilization packaged in a third capsule (CP3), d. a biologically derived bio -pesticide capably effecting crop protection, packaged in a fourth capsule (CP 4) e. a biologically derived bio-fungicide capably effecting crop-protection, packaged in a fifth capsule (CP5) f. a biologically derived bio-nematicide capably effecting crop- protection, packaged in a sixth capsule (CP6) g. a biologically derived bio-foliar protector capably effecting crop- protection, packaged in a seventh capsule (CP7) h. a biologically derived bio-nutrient capably effecting nutrition for microbes and crop, packaged in an eighth capsule (CP8) said first bio-fertilizer comprising combination of Azotobacter chroococcum, Azotobacter vinelindii, Azotospirillum lipoferum, Rhizobium, Acetobacter aceti, Azospirillium spp. ; said second bio -fertilizer comprising combination of Pseudomonas putida and Aspergillus niger or Bacillus megaterium; said third bio-fertilizer comprising Frateuria aurentina; said bio-pesticide comprising Pseudomonas fluroscence; said bio-fungicide comprising Trichoderma viridae or Trichoderma hurzianum; said bio - nemuccide comprising Pacilomyces liliacinus and Bacillus subtilis; said bio-foliar protector comprising combination of Beauveria basssiana, Metarhizium anisoplae and Verticillium anosoplae; said bio-nutrition comprising combination of Bacillus subtilis, Bacillus polymyxa, Pseudomonas fluorescence and Pseudomonas putida; wherein, a water soluble compound PHH is used as a base which is impregnated with the relevant microbes prior to each capsulization, for later - packaging the relevant impregnated base. 2) A capsulated formulation with beneficial microbes, as claimed in claim 1, wherein the capsules are of various types including vegetable capsules, gelatin capsules, soft capsules, or gel capsules. 3) A capsulated formulation with beneficial microbes, as claimed in claim 1, wherein the capsules are of various sizes including 000, 00, 0, 1, 2, 3 or 4. 4) A capsulated formulation with beneficial microbes, as claimed in claim 1, wherein in each of said capsules, quantity of each of said microbes, packaged therein, has equal proportion by weight. 5) A capsulated formulation with beneficial microbes, as claimed in claim 1, wherein a first group of the packaged capsules with relevant contents packaged therein, comprising said CP1, CP2, CP3 and CP8 is provided to the soil at the start of cultivation and also afterwards as and when required; a second group of the capsules with relevant contents packages therein, comprising said CP4, CP5, CP6 and CP8 is provided to the soil after 3 to 4 days of cultivation and also afterwards as and when required; and a third group of the capsules with relevant contents packaged therein, comprising said CP7 and / or CP8 is dissolved in water capably forming a protective solution which is sprayed on grown foliage for foliar protection. 6) A capsulated formulation with beneficial microbes, as claimed in claim 1, wherein the material used for making each of said capsules is soluble in water, capably working as additional carbon source for said microbes and the soil, when dissolved in soil-water. 7) A capsulated formulation with beneficial microbes as claimed in claim 1, wherein all different microbes packaged in each of said capsules are in dormant state, thereby capably maintaining compatibility of said microbes in each of said capsules. 8) A method of preparation of a capsulated formulation with beneficial microbes for healthy plant growth, comprising the steps of: a. getting ready a required predetermined first quantity of nutrient broth used for bacteria- medium needed for preparation of corresponding required quantity of any one formulation from a group -A of formulations of claim 1 comprising; i. said first bio -fertilizer, ii. said second bio-fertilizer, iii. said third bio-fertilizer, and iv. said bio-pesticide, v. said bio-nutrition, or getting ready a predetermined second quantity of a potato dextrose broth used for fungus-medium needed for preparation of corresponding required quantity of any one formulation from a group -B of claim 1 comprising; vi. said bio-fungicide, vii. said bio-nemuccide, and viii. said bio-foliar protector b. charging distilled water into a conical flask; c. charging the required relevant broth of step (a) into said flask at a predetermined first temperature; d. autoclaving said flask with contents thereof, at a predetermined second temperature for a predetermined first duration; e. inoculating in said flask of step (d) relevant microbe under aseptic conditions at a predetermined third temperature and repeating the step of inoculating separately for each strain in separate corresponding flask; f. transferring each of said flasks of step (e) separately to a shaker; g. maintaining each of the transferred flasks on said shaker at a predetermined fourth temperature for a predetermined second duration in case of said flask containing nutrient broth or at a predetermined seventh temperature for a predetermined fifth duration in case of said flask containing potato dextrose broth; h. transferring said flask to an incubator after completion of step (g); i. incubating contents of said flask at a predetermined fifth temperature for a predetermined third duration in case of said flask containing nutrient broth or at said predetermined fifth temperature for a predetermined sixth duration in case of said flask containing potato dextrose broth; j. adding to said flask of step (i), SMS of a predetermined first concentration; k. mixing powder base of said PHH of claiml, with contents of said flask of step (j) to form relevant formulation from said group (A) or group (B) of step (a), to deliver a predetermined second concentration of broth by weight in the mixture with a predetermined first potency;

1. drying the formulation - mixture of step (k) at a predetermined sixth temperature for a predetermined fourth duration and mixing the dried products;

m. packing a predetermined required quantity of the dried and mixed material of step (1) in a predetermined capsule of predetermined required size, of predetermined shape, said capsule being made of a predetermined material.

9) A method of preparation of a capsulated formulation, as claimed in claim 8 wherein,

a. a range of said first temperature is from 18°C to 37°C, preferably from 20°C to 32°C, and more preferably from 25°C to 30°C;

b. a range of said second temperature is from 74°C to 125°C, preferably from 94°C to 124°C and more preferably from 115°C to 121°C;

c. a range of said third or fifth or seventh temperature is from 10°C to 35°C, preferably from 15°C to 32°C more preferably from 25°C to 30°C; d. a range of said fourth temperature is from 10°C to 35°C, preferably from 15°C to 32°C, more preferably from 25°C to 30°C;

e. a range of said sixth temperature is from 20°C to 37°C, preferably from 25°C to 35°C, more preferably between 27°C to 32°C; f. a range of said first duration is from 20 minutes to 90 minutes preferably from 25 minutes to 60 minutes and more preferably from 30 minutes to 45 minutes;

g. a range of said second or third duration is from 1 day to 7 days, preferably from 2 days to 6 days, more preferably from 3 days to 4 days; h. a range of said fourth duration is from 5 hours to 24 hours, preferably from 6 hours to 18 hours and more preferably from 7 hours to 10 hours; i. a range of said fifth or sixth duration is from 3 days to 10 days preferably from 4 days to 8 days, and more preferably from 5 days to 7 days;

j. a range of said first concentration of SMS by weight is from 1% to

5%, preferably from 2% to 4% and more preferably from 2.5% to 3.5% of weight of broth;

k. a range of said second concentration of broth by weight is from 5% to 10%, preferably from 6% to 9% more preferably from 7% to 8% of weight of mixture of broth and PHH;

1. a range of said first potency is from 1 x 10^~7 to 5 x 10^~7, preferably from

2 x 10^"7 to 4 x 10^"7, and more preferably from 2 x 10^"7 to 3 x 10^"7;

m. a range of said predetermined required quantity of said dried material is from 0.1 gm to 2 gm, preferably from 0.5 gm to 1.5 gm, more preferably from 0.8 gm to 1.2 gm;

n. A range of said predetermined size is selected from the group

consisting of 000, 00, 0, 1, 2, 3, 4 and such other sizes;

o. A range of said predetermined shapes is selected from the group

consisting of appropriate shapes;

p. A range of said predetermined material is selected from the group consisting of vegetable capsules, gelatine capsules, soft capsules, gel capsules , and such other capsules.