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Field of Invention:
The invention relates to a formulation for controlling mealy bugs and other pests pathogens on plants and in soil using a consortium of microorganisms. The composition consists mainly spore forming microorganisms belonging to bacillus species.
Background of the Invention:
Mealy bug is a serious pest of many plants including grape, citrus, fig, guava, mango, vegetables, etc, yield losses can be up to 50 percent due to meaty bug damage. The most common regions that are infested in plant are the leaves and tender twigs. They also attack the fruits, flowers and even hardier stems. They injure the plant by sucking sap, as a result of which tender twigs and leaves are deformed and stunted in growth. Such injury will seriously effect the growth and yield of the crop. Besides their excrete attracts ants and sooty mould, a fungus. Incase of heavy deposit of the fungus photosynthetic activity of the plant is affected resulting in leaf fall and ultimate death of the plant.
Mealy bugs are hard-to-kill insects as the chemical insecticides are not very effective due to the mealy coating on the insects. The invention relates to the use of consortium for agriculture and horticulture field for killing, controlling or otherwise adversely impacting the life cycle of pests & pathogens on plants and in the soil.
Pesticides are an important component throughout the world. Chemical agents include insecticides for the control of insect pests and fungicides for control of plant & soil pathogens. It is important to control pests and pathogens that affect the crops and inflict major damage resulting in crop loss. While several chemicals have proved to be effective for controlling pests & pathogens they have also
contributed to an unacceptable environmental damage that directly affects human health.
Since traditional insecticides work on the principle of chemical toxicity, the insects are capable of developing resistance to the toxins through genetic mutations. It had been observed that several pests, which are susceptible to certain chemical compounds, do not get killed even with higher doses.
Most chemicals need to be used with extreme caution. Direct exposure to such chemicals can lead to human health damage and prolonged exposure may lead to toxic shock and even death.
The chemical insecticides/ fungicides are designed to leave behind a toxic residue either on the plants or in the soil.
Certain chemical insecticides also cannot destroy insects like mealy bugs, which are present below the bark of the plant and also cannot penetrate the mealy covering.
US patent 6638910 and related patents talk about a composition for controlling the pests. The said composition relates to an antibiotic producing and metabolite producing Bacillus subtilis strain that exhibits insecticidal, antifungal and antibacterial activity. The composition for protecting or treating plants and fruit from fungal and bacterial infections and corn rootworm infestations comprising effective amounts of an Iturin A, a plipastatin, a surfactin and an agrastatin. Also included in the invention is a solvent extractable, small molecular weight (<10,000 daltons) com rootworm-active metabolite produced in the supernatant. Further the said patent teaches method of protecting or treating plants from fungal and bacterial infections and com rootworm infestations.
However the disadvantage the composition of the said patent is firstly it is not effective in the controlling the mealy bugs and other plant pests. Secondly it involves application of harmful chemicals.
US patent 6635245 and related patents disclose a strain of bacillus for controlling plant diseases. The antibiotic-producing Bacillus sp. is provided that exhibits antifungal activity only on certain specific plant pathogens and no antibacterial activity is provided by said invention. Also discloses a method of treating or protecting plants, fruit and roots from fungal infections by applying an effective amount of these strains, supematants produced by these strains or a metabolite isolated from these strains. The invention further encompasses the synergistic fungicidal effect of using a strain of NRRL Accession Nos. B-30087 together with B-21661 (AQ 713).
The disadvantage the composition of the US patent 6635245 is that it is effective only in the treatment of the fungal infection in a plant.
Therefore the object of to obviate the disadvantages of the prior art and to provide a to provide a safe method for controlling mealy bugs and other pests and pathogens using a consortium of microorganisms.
Object of the Invention:
As discussed earlier the object of the present invention is to provide a safe method for controlling mealy bugs and other pests and pathogens using a consortium of microorganisms.
Another object of the present invention to provide a method for controlling insect populations that is safe and convenient to use.
Yet another object of the invention is to provide a method for controlling insect pests and pathogens, which reduces worker exposure to hazardous and toxic compounds.
Further, object of the present invention is to provide a method of lowering the pH of the soil for enhancing the nutrient uptake and also controlling the root and soil borne pathogens.
It is yet another object of the present invention to provide a method of protecting plants from insect infestations through the entire life cycle of the plant.
Summary of the invention:
The present invention relates to a formulation for controlling mealy bugs and other pest pathogens comprising a consortium of microorganisms belonging to Bacillus species and a carrier.
The synergistic action of all the microorganisms is resulting in the effective control. The said formulation is for foliar application on plants, soil applications to enhance the nutrient uptake and increasing the growth and yields of the plants, for soil application for controlling root and soil borne pathogens and pests. The formulation is also for the control of post harvest berry / fruit rot.
The said formulation is also effective in controlling several other insects and pathogens like nematodes, powdery mildew, downy mildew, wooly aphids Fusarium, Alternaria, Anthracnose, Rhizoctonia, Pythium, Botrytis, Xanthomonas, thrips, mites, beetles, bollworms, sodoptera, heliothis, caterpillars, nematodes, termites and diseases like root rot, fruit rot, stem rot, blight & rust etc
The microorganisms belonging to Bacillus species are selected form the group consisting of Bacillus megaterium, Bacillus pumilus, Bacillus poiymyxa, Bacillus
subtilis, Bacillus mecentricus, Bacillus firmus, Bacillus coagulans, Bacillus lichenifbrmus, Bacillus lateropsoruss and Bacillus amyloliquificans.
The carrier used in the invention is selected from dextrose, lactose, talc, and malto-dextrine starch. However, preferred carrier is dextrose.
The formulation is in a powder, semi solid, gel, solid or liquid form.
The formulation of the present invention can be applied both on the foliar parts of plan and soil.
Further, the invention relates to the method of preparing the formulation comprising the steps of:
maintaining the selected microorganism as pure slant cultures wherein
each culture is manufactured individually in fermenters,
harvesting the broth from the fermenters,
centrifuging the said broth to obtain a biomass,
spray drying the biomass, and
blending the calculated amount of each culture in a Double Cone Blender along with the carrier to obtain the formulation.
The pure culture is manufactured in fermenters by standard operating procedures.
The formulation is useful in controlling powdery mildew downy mildew, wooly aphids fusarium, altenaria, anthracnose, rhizoctonia, pythium, botrytis, thrips, xanthomonas, nematodes, beetles, bollworms, sodoptera, heliothis, mites, caterpillars, termites and diseases like root rot, fruit rot, stem rot, blight & rust.
Further, the formulation is useful in killing the mealy bugs and also cleaning up the residues selected by the mealy bugs and residues of the dead bugs.
The application of the formulation on soil is better nutrient uptake and increase in the growth and yield of plant.
The present invention further provide the method for controlling mealy bugs and other pest pathogens comprising the steps of:
preparing a solution by mixing 1-5 gm of formulation in one litre of water, spraying the said solution over the plant parts or soil.
Details of the invention:
The use of the formulation containing the consortium of microorganisms has been found to be very effective in controlling various pests and pathogens on plants and in soil. The use of environment friendly biological control is a better choice against chemical pesticides. It has been found that the use of the consortium of present invention for spraying on plants and soil has effectively controlled mealy bugs, powdery mildew, downy mildew, nematodes, other insects and pathogens. Trials have also indicated that it can effectively control post harvest rot of fruits / vegetables.
The effect of the probiotic microorganisms present in the consortium results from competitive inhibition / competitive exclusion of pathogens by competing for the space and food source with the pathogens and pests, production of acids and other extra cellular products possessing anti microbial activity.
production of substances / enzymes inhibitory to the growth of pests and pathogens
Synergistic action of the various microorganisms present is due to one or more actions referred above. The mode of inhibition / insecticidal / fungicidal action varies from pest to pest and pathogen to pathogen. The extra cellular compound / enzyme harmful to one kind of pest / pathogen may not be harmful to another, there the effective result is due to the preemptive consumption of food sources of the pest / pathogen present in the formulation.
The formulation when applied to soil helps in fighting the root and soil borne pathogens and pests and also helps in enhancing the nutrient uptake by lowering the pH of the soil. This is a very important aspect as the pH of soil at most of the places is becoming alkaline due to the indiscriminate use of chemical fertilizers.
Composition:
The formulation contains a consortium of organisms belonging mainly to bacillus spp. in a dextrose carrier. The said formulation contains a strength of up to 200 billion organisms per gram (200 billion cfu/gram). The microorganisms used are mainly from the bacillus species and are from a group of microorganisms consisting of Bacillus megaterium, Bacillus pumilus, Bacillus polymyxa, Bacillus subtillis, Bacillus mecentricus, Bacillus firmus, Bacillus coagulans, Bacillus licheniformus, Bacillus lateropsorus Bacillus amyloliquificans.
The carrier used can also be any other carrier like dextrose like lactose, talc, maltodextrine, starch etc.
The formulation can be in a powder, semi solid, gel, solid or liquid form. Probiotics:
The term Probiotic originated from two Greek words meaning 'for life' and contrasted with the term Antibiotic, which means 'against life'.
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Examples of beneficial groups of microorganisms are those, which inhabit the gastrointestinal tract of animals and larger ecosystems like soil and water bodies.
Though the term 'Probiotics' is generally used with reference to animal ecosystem, the term is increasingly used in context of other ecosystems like soil, plants, water bodies etc.
Probiotics in relation to plants and agriculture can be for various applications the, main aim being health of the plants avoiding the use of harmful fungicides and insecticides.
The ideal probiotic possess the following features: should be non-toxic beneficial to the host in some way of high viability
stable on storage and in the field
able to survive in or colonize on the intended host
Bacillus species:
The word bacillus is a descriptive term for the appearance of certain bacteria when viewed microscopically. It derives from the Latin for "staff" and means "rod- shaped". Bacillus thus is the name given to a genus of rod-shaped, Gram positive, aerobic, spore-forming bacteria belonging to the family Bacillaceae.
The bacteria of the genus Bacillus is a ubiquitous group of bacteria commonly recovered from water, soil, air, and decomposing plant residue. The bacteria of genus Bacillus produce endospores, which can endure extreme conditions of heat and desiccation in the environment. Bacillus species produce a variety of proteases and other enzymes that enable it to degrade a variety of natural
substrates and contribute to nutrient cycling. However, under most conditions the organism is not biologically active but exists in the spore form (Alexander, 1977). Majority species of Bacillus are considered benign organisms, as they do not possess traits that cause disease and are saprophytic. They are not considered pathogenic or toxigenic to humans, animals, or plants. The potential risk associated with the use of this bacterium in fermentation facilities is low. Bacillus bacteria are ubiquitous in nature and can form roughly spherical spores when conditions are stressful in order to survive in a dormant state for extended periods. There exist a huge number of species in this genus, most of which are harmless.
Bacillus species are gram positive and catalase positive bacteria that use oxygen as their terminal electron acceptor in their energy metabolic pathway. Though they are aerobic, many species are facultative to microaerophilic, which gives them the advantage to grow and proliferate under low oxygen conditions also. Individual Bacillus appears as rods under the microscope with usually a substantial number of the rods having an oval endospore which tends to bulge the bacterium at one end (terminal spore) or in the middle (central spore). The endospore is a dormant stage in the life cycle of Bacillus bacteria, which is formed due to adverse environmental conditions like lack of nutrients, lack of oxygen etc. Endospore is a natural encapsulation of the bacterial cell to allow it to vade over harsh conditions of nature and survive changing microecological conditions better. Spores can tide unfavorable conditions and germinate when favorable conditions arise.
On provision of favorable conditions like nutrients, water the spore coats imbibe water, swell, and the increased water content causes a rise in the metabolic rate of the sporulated bacilli. Outgrowths begin to protrude from the spore-coats and the spores germinate and transform into viable vegetative cells. Usually, germination takes place about four hours after the provision of favorable culture conditions. These vegetative cells proliferate on the nutrients multiplying rapidly, producing organic acids and other metabolites as long as the nutrients are available.
Application of the formulation: Foliar Application:
1mg - 5 grams of this formulation depending on the strength is dissolved per litre of water and sprayed using a spray pump on the stems, branches, leaves, fruits and soil.
Soil Application:
This formulation can be applied to soil by dissolving it in water and applying through drip irrigation or drenching at the rate of 1 kg dissolved in sufficient water for 1 acre. Lower application doses will be required if the product is of higher concentration.
Manufacturing Process:
The process of manufacturing of various microorganisms is as follows.
all the selected species are maintained as pure slant cultures in the microbiological laboratory by qualified and experienced microbiologists, each culture is manufactured individually and separately as pure culture in fermenters as per the Standard Operating Procedure, the broth from the fermenters is harvested and centrifuged to obtain a biomass and the biomass is spray dried.
the spray-dried powder of each culture is analyzed and preserved separately.
Calculated amount of each culture is weighed and added to a Double Cone Blender along with prescribed quantity of Dextrose Monohydrate, which is the carrier and blended.
the blended product is analyzed for the active ingredients as per the formula.
the product is finally packed in polyethylene bags and then in plastic containers as per the order.
Description of the individual steps of production: Strains:
The bacterial strains of Bacillus species are Gram-positive spore forming rods and produce organic acids during their growth.
Characteristic of the producing strains:
The bacterial strains are Gram positive spore forming Bacillus species with terminal spores, aerobic to microaerophilic in nature. They produce various organic acids and other metabolites specific to each strain. The growth temperature range is of 25 to 50 °C. Optimum pH is 5.5 to 8.0.
Isolation and storage:
Pure cultures are isolated and their ability to produce organic adds and spore formation is checked and selected cultures are stored in refrigerator.
Inoculum preparation:
The culture, which is previously selected, is inoculated into flasks containing seed medium. The inoculum is cultivated on a shaker for about 24 hours at suitable temperature specific to the culture, generally between 25 to 50 oC. The inoculum is also checked for its purity in a phase contrast microscope before inoculating into a batch.
Fermentation:
The fermentation medium consisting of nutrients specific to the strain is constituted in the fermenter, the pH is adjusted specific to the strain and the medium is sterilized and cooled. When required temperature is attained, the seed culture is inoculated into the fermenter asepticaliy.
Cultivation runs at the required temperature specific to the strain (25 to 50 °C) for about 36-48 hours. Samples are drawn asepticaliy from the fermenter periodically and the pH is measured and microscopic observation is done. After
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completion of growth and attaining maximum sporulation the batch is harvested by centrifugation.
Centrifugation:
After completion of the fermentation the batch is harvested, the broth is removed through the harvest line and centrifuged to obtain biomass.
Spray drying:
The biomass that is obtained in the centrifugation is spray dried to obtain concentrated spray dried Powder. The spray dried powders of different strains are analyzed separately and preserved separately.
Blending:
Calculated amount of spray-dried powder of each culture is weighed and blended with required quantity of dextrose monohydrate to get a premix of the product. The premix is labeled and analyzed for the active ingredients as per the standard procedures and preserved till further use. Calculated amount of the premix is weighed and blended with required quantity of dextrose monohydrate which is the carrier, in a double cone blender as per the Standard Operating Procedure, to get the final product. The final product is labeled and analyzed for the active ingredients as per the Standard Procedures and preserved.
Packaging:
On completion of the analysis and approval, the product is finally packed in polythene bags and then in plastic containers as per the order.
The formulation was tested in various field trials and the examples are given below.
Example 1:
A solution made by mixing 3 gms of the formulation containing the said consortium of organisms per litre of water was sprayed over grape plants infected with mealy bugs. It was observed that the mealy bugs were dead and also the mealy secretion on the berries was not found.
The grape farm was divided into four parts as under:
• Control
• Sprayed with water
• Sprayed with dextrose (3 gm per litre of water)
• Sprayed with the said formulation (3gm per litre of water)
• Sprayed with chemical insecticide
There was no difference observed in the control of mealy bugs in part no. 1, 2 & 3 after 4 days.
Part no. 4 treated with the formulation containing the said consortium of microorganisms resulted in the maximum destruction of mealy bugs compared to the chemical insecticide.
It was observed that the mealy bugs under the bench were also dead in the plants treated with the said formulation where as the chemical insecticides did not effect the bugs under the bark.
Example 2:
The formulation was used for improving the shelf life of grapes through control of post harvest berry lots.
The bio efficacy study conducted at a National Research Institute concluded that
A pre harvest spray at 2 grams to 8 grams of the formulation per litre of water reduced the number of fallen berries.
Post harvest berTy rolling by unidentified yeast and bacteria was not present in the packing for local market & exports. No phytotoxicity symptom of any kind was observed.
The treated berries also showed significantly better freshness ratings over the controls.
Example 3:
Repeated oral toxicity tests have indicated that the formulation is non-toxic, non- virulent and safe for use.
Example 4:
The other field trials have indicated that the formulation has effectively controlled pathogens like powdery mildew, wooly aphids, nematodes, termites, fusarium, downy mildew, anthramose, thrips, mites & beetles.
The field trials were conducted on grapes, citrus, sugarcane, gerbera, pomegranate, custard apple, guava, mango etc.
Example 5:
The formulation dissolved in water and applied to soil after twenty-four hours lowered the pH in the rhizosphere from 8 to 5.
Conclusion:
1 mg - 5 grams of this formulation per liter of water sprayed over the entire plant and applied to soil by drenching controls / mealy bugs wooly aphids,powdery mildew, downy mildew, fusarium, alternaria, anthracnose, rhizoctonia, pythium, botrytis, thrips, xanthomonas, nematodes, beetles, bollworms, sodoptera, heliothis, mites, caterpillars, termites and diseases like root rot, fruit rot, stem rot, blight & rust, mealy bugs, powdery mildew, downy mildew, fusarium, anthracanose, die back, nematodes, termites and other pests and pathogens. The application in soil also helps in higher growth and yield of the plants as several nutrients are made available to the plants as the microorganisms lower the pH of the soil. The application in soil controls root and soil borne pathogens and increases the root growth. The said formulation is non-toxic and does not leave behind toxic residues. Lower doses are required if the formulation used is of a higher concentration.

We claim:
1. A formulation for controlling mealy bugs and other pest pathogens comprising a consortium of microorganisms belonging to Bacillus species and a carrier.
2. The formulation as claimed in claim 1 wherein the microorganisms are from the Bacillus species.
3. The formulation as claimed in claim 1 wherein formulation contains up to 200 billion organisms per gram (200 billion cfu/gram).
4. The formulation as claimed in claim 1 wherein the microorganisms are selected from the group consisting of Bacillus megaterium, Bacillus pumilus, Bacillus polymyxa, Bacillus subtilis. Bacillus mecentricus, Bacillus firmus, Bacillus coagulans, Bacillus licheniformus, Bacillus lateropsoruss and Bacillus amyloliquificans.
5. The formulation as claimed in claim 1 wherein the carrier is dextrose, lactose, talc, and maltodextrine starch.
6. A formulation as claimed in claim 1 wherein the preferable carrier is dextrose.
7. The formula as claimed in claim 1 wherein the formulation is in a powder, semi solid, gel, solid or liquid form.
8. A formula as claimed in claim 1 wherein the formulation is applied both on the foliar parts of plan and soil.
9. The method for controlling mealy bugs and other pest pathogens as claimed in claim 1 comprising the steps of:
preparing a solution by mixing 1-5 gms of formulation in one liter of water.
spraying the said solution over the plant parts or soil.
10. The method of preparing the formulation as claimed in claim 1 comprising the steps of:
maintaining the selected microorganism as pure slant cultures wherein each culture is manufactured individually in fermenters, harvesting the broth from the fermenters, centrifuging the said broth to obtain a biomass, spray drying the biomass, and
blending the calculated amount of each culture in a Double Cone Blender along with the carrier to obtain the formulation.
11. The method of preparing the formulation as claimed in claim 9 wherein the pure culture is manufactured in fermenters by standard operating procedures.
12. The formulation as claimed in claim 1 wherein the said formulation is using powdery mildew downy mildew, wooly aphids fusarium, altenaria, anthracnose, rhizoctonia, pythium, botrytis, thrips, xanthomonas, nematodes, beetles, bollworms, sodoptera, heliothis, mites, caterpillars, termites and diseases like root rot, fruit rot, stem rot, blight & rust.
13. The formulation as claimed in claim 1 wherein the said formulation is useful in killing the mealy bugs and also cleaning up the residues selected by the mealy bugs and residues of the dead bugs.
14. A formulation for controlling mealy bugs and other pest patrogens substantially as herein described with reference to the foregoing examples.
15. The method for controlling mealy bugs and other pest pathogens substantially as herein described with reference to the foregoing examples.
16. The method of preparing the formulation substantially as herein described with reference to the foregoing examples.