DESC:FIELD
The present disclosure relates to a pesticidal formulation and a process for its preparation. Particularly, the present disclosure relates to a pesticidal formulation against pests of family Pseudococcidae.
BACKGROUND
The background information herein below relates to the present disclosure but is not necessarily prior art.
Pseudococcidae is a family of hemipterous insects (suborder Homoptera) comprising mealybugs. Mealybug females feed on plant sap, normally in roots or other crevices, and in a few cases the bottoms of stored fruit. They attach themselves to the plant and secrete a powdery wax layer (hence the name "mealy" bug) that is used for protection while they suck the plant juices. Mealybugs can infest a number of indoor houseplants, particularly tropical species. Mealybugs can pose a serious threat to some commercial crops, such as mango fruits, citrus fruits and the like. Currently, the infected plants are treated with the chemicals such as permethrin, bifenthrin or cyfluthrin. Early life pesticide exposure to permethrin insecticide in human beings, has long-term consequences leading to cardiac hypotrophy, increased calcium and Nrf2 gene expression levels in old age. When bifenthrin gets on the skin of human beings, it can cause tingling, itching, burning or numbness at the site of contact, act as irritant for nose, throat, and lungs.
There is, therefore, felt a need to develop a pesticidal formulation that mitigates the drawbacks mentioned herein above or at least provides a useful alternative.
OBJECTS
Some of the objects of the present disclosure, which at least one embodiment herein satisfies, are as follows:
It is an object of the present disclosure to ameliorate one or more problems of the prior art or to at least provide a useful alternative.
An object of the present disclosure is to provide a pesticidal formulation.
Another object of the present disclosure is to provide a pesticidal formulation which comprises a bacterial concentrated extract.
Still another object of the present disclosure is to provide a pesticidal formulation which comprises B.velezensis extract and P.megaterium extract.
Yet another object of the present disclosure is to provide a pesticidal formulation which has a pesticidal activity against mealy bugs.
Still another object of the present disclosure is to provide a pesticidal formulation which is non-phytotoxic.
Yet another object of the present disclosure is to provide a pesticidal formulation that exhibits 100% solubility in a fluid medium.
Still another object of the present disclosure is to provide a process for the preparation of a pesticidal formulation.
Still another object of the present disclosure is to provide a process for the preparation of a pesticidal formulation which is simple, economic and environment friendly.
Other objects and advantages of the present disclosure will be more apparent from the following description, which is not intended to limit the scope of the present disclosure.
SUMMARY
The present disclosure relates to a pesticidal formulation comprising a predetermined amount of at least one bacterial concentrated extract, a predetermined amount of a natural repellent, a predetermined amount of a penetrating agent, a predetermined amount of a surfactant, a predetermined amount of a stabilizing agent, optionally a predetermined amount of a flavouring agent and a predetermined amount of a fluid medium.
The predetermined amount of the predetermined amount of the bacterial concentrated extract is in the range of 2 mass% to 10 mass%, the predetermined amount of the natural repellent is in the range of 1 mass% to 5 mass%, the predetermined amount of the penetrating agent is in the range of 0.5 mass% to 5 mass%, the predetermined amount of the surfactant is in the range of 0.01 mass% to 2 mass%, the predetermined amount of the stabilizing agent is in the range of 0.01 mass% to 3 mass%, the predetermined amount of the flavouring agent is in the range of 0 mass% to 1 mass%; and the predetermined amount of the fluid medium is in the range of 75 mass% to 97 mass%, wherein the mass% of each ingredient is with respect to the total amount of the formulation.
The bacterial concentrated extract is obtained from at least one bacterial species selected from the group consisting of Bacillus species and Priestia species.
The Bacillus species is selected from the group consisting of Bacillus velezensis, Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus aryabhattai and the Priestia species is Priestia megaterium.
The bacterial concentrated extract is extract of a combination of Bacillus velezensis and Priestia megaterium.
The natural repellent is selected from the group consisting of neem oil, castor oil, lemon grass oil and eucalyptus oil. The penetrating agent is selected from the group consisting of isopropyl alcohol, ethanol and methanol. The surfactant is selected from the group consisting of anionic surfactants, non-ionic surfactants, cationic surfactants and zwitterionic surfactants. The stabilizing agent is selected from the group consisting of sodium carbonate, sodium carbonate, potassium bicarbonate and sodium bicarbonate. The flavouring agent is selected from the group consisting of lemon grass oil, and eucalyptus oil and the fluid medium is water.
The anionic surfactant is selected from the group consisting of sodium laureth sulfate, sodium lauryl sarcosinate, sodium myreth sulfate, sodium pareth sulfate, sodium stearate, sodium lauryl sulfate, a-olefin sulfonate and ammonium laureth sulfate. The non-ionic surfactant is selected from the group consisting of polyethylene glycol, glycerol, sorbitan monooleate, ethylene oxide condensate of alkyl phenol and polysorbate 80, the cationic surfactant is selected from the group consisting of benzethonium chloride, benzododecinium bromide ethyl lauroyl arginate, dimethyl dioctadecyl ammonium chloride, dioleoyl-3-trimethylammonium propane. The zwitterionic surfactant is selected from the group consisting of betaines, sulfobetaines, and amine oxides.
In an embodiment of the present disclosure, the formulation is in the form of a liquid.
Further, the present disclosure relates to a process for the preparation of a pesticidal formulation. The process comprises mixing predetermined amounts of at least one bacterial concentrated extract, a natural repellent, a penetrating agent, a surfactant, a stabilizing agent, optionally a flavouring agent in a predetermined amount of fluid medium under stirring for a predetermined time period at a predetermined temperature to obtain the pesticidal formulation.
The predetermined time period is in the range of 15 minutes to 60 minutes and the predetermined temperature is in the range of 25 oC to 35 oC.
The natural repellent is selected from the group consisting of neem oil, castor oil, lemon grass oil and eucalyptus oil, the penetrating agent is selected from the group consisting of isopropyl alcohol, ethanol and methanol, the surfactant is selected from the group consisting of anionic surfactants, non-ionic surfactants, cationic surfactants and zwitterionic surfactants, the stabilizing agent is selected from the group consisting of sodium carbonate, potassium bicarbonate and sodium bicarbonate, the flavouring agent is selected from the group consisting of lemon grass oil and eucalyptus oil and the fluid medium is water.
The anionic surfactant is selected from the group consisting of sodium laureth sulfate, sodium lauryl sarcosinate, sodium myreth sulfate, sodium pareth sulfate, sodium stearate, sodium lauryl sulfate, a-olefin sulfonate and ammonium laureth sulfate, the non-ionic surfactant is selected from the group consisting of polyethylene glycol, glycerol, sorbitan monooleate, ethylene oxide condensate of alkyl phenol and polysorbate 80. The cationic surfactant is selected from the group consisting of benzethonium chloride, benzododecinium bromide ethyl lauroyl arginate, dimethyl dioctadecyl ammonium chloride and dioleoyl-3-trimethylammonium propane and the zwitterionic surfactant is selected from the group consisting of betaines, sulfobetaines, and amine oxides.
The predetermined amount of the bacterial concentrated extract is in the range of 2 mass% to 10 mass%, the predetermined amount of the natural repellent is in the range of 1 mass% to 5 mass%, the predetermined amount of the penetrating agent is in the range of 0.5 mass% to 5 mass%, the predetermined amount of the surfactant is in the range of 0.01 mass% to 2 mass%, the predetermined amount of the stabilizing agent is in the range of 0.01 mass% to 3 mass%, the predetermined amount of the flavouring agent is in the range of 0 mass% to 1 mass% and the predetermined amount of the fluid medium is in the range of 75 mass% to 97 mass%, wherein the mass% of each ingredient is with respect to the total amount of the formulation.
The pesticidal formulation is used against pests of family Pseudococcidae.
BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWING
The present disclosure will now be described with the help of the accompanying drawing, in which:
Figure 1(A) illustrates image of mealy bugs infested plant leaves before treatment with the commercially available pesticidal formulation, (B) images of mealy bugs infested plant leaves after treatment with the commercially available pesticidal formulation, (C) illustrates images of mealy bugs infested plant leaves before treatment with a pesticidal formulation in accordance with the present disclosure, (D) illustrates images of mealy bugs infested plant leaves after treatment with a pesticidal formulation in accordance with the present disclosure, (E) illustrates microscopic images of mealy bugs infested plant leaves after treatment with the commercially available pesticidal formulation, (F) illustrates microscopic images of mealy bugs (which turned dark brown color indicating mortality of the mealy bugs) infested plant leaves after treatment with a pesticidal formulation in accordance with the present disclosure.
DETAILED DESCRIPTION
The present disclosure relates to a pesticidal formulation and a process for its preparation. Particularly, the present disclosure relates to a pesticidal formulation against pests of family Pseudococcidae.
Embodiments, of the present disclosure, will now be described with reference to the accompanying drawing.
Embodiments are provided so as to thoroughly and fully convey the scope of the present disclosure to the person skilled in the art. Numerous details are set forth, relating to specific components, and methods, to provide a complete understanding of embodiments of the present disclosure. It will be apparent to the person skilled in the art that the details provided in the embodiments should not be construed to limit the scope of the present disclosure. In some embodiments, well-known processes, well-known apparatus structures, and well-known techniques are not described in detail.
The terminology used, in the present disclosure, is only for the purpose of explaining a particular embodiment and such terminology shall not be considered to limit the scope of the present disclosure. As used in the present disclosure, the forms "a,” "an," and "the" may be intended to include the plural forms as well, unless the context clearly suggests otherwise. The terms "comprises," "comprising," “including,” and “having,” are open ended transitional phrases and therefore specify the presence of stated features, integers, steps, operations, elements, modules, units and/or components, but do not forbid the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof. The particular order of steps disclosed in the method and process of the present disclosure is not to be construed as necessarily requiring their performance as described or illustrated. It is also to be understood that additional or alternative steps may be employed.
As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed elements.
The terms first, second, third, etc., should not be construed to limit the scope of the present disclosure as the aforementioned terms may be only used to distinguish one element, component, region, layer or section from another component, region, layer or section. Terms such as first, second, third etc., when used herein do not imply a specific sequence or order unless clearly suggested by the present disclosure.
Pseudococcidae is a family of hemipterous insects (suborder Homoptera) comprising mealybugs. Mealybug females feed on plant sap, normally in roots or other crevices, and in a few cases the bottoms of stored fruit. They attach themselves to the plant and secrete a powdery wax layer (hence the name "mealy" bug) that is used for protection while they suck the plant juices. Mealybugs can infest a number of indoor houseplants, particularly tropical species. Mealybugs can pose a serious threat to some commercial crops, such as mango fruits, citrus fruits and the like. Currently, the infected plants are treated with the chemicals such as permethrin, bifenthrin or cyfluthrin. Early life pesticide exposure to permethrin insecticide in human beings, has long-term consequences leading to cardiac hypotrophy, increased calcium and Nrf2 gene expression levels in old age. When bifenthrin gets on the skin of human beings, it can cause tingling, itching, burning or numbness at the site of contact, act as irritant for nose, throat, and lungs.
The present disclosure provides a pesticidal formulation and a process for its preparation.
In an aspect, the present disclosure provides a pesticidal formulation for mealy bugs.
In accordance with the embodiment of the present disclosure, the pesticidal formulation for mealy bugs comprises a predetermined amount of at least one bacterial concentrated extract, a predetermined amount of a natural repellent, a predetermined amount of a penetrating agent, a predetermined amount of a surfactant, a predetermined amount of a stabilizing agent, optionally a predetermined amount of a flavouring agent and a predetermined amount of a fluid medium.
In accordance with an embodiment of the present disclosure, the predetermined amount of the bacterial concentrated extract is in the range of 2 mass% to 10 mass%, the predetermined amount of the natural repellent is in the range of 1 mass% to 5 mass%, the predetermined amount of the penetrating agent is in the range of 0.5 mass% to 5 mass%, the predetermined amount of the surfactant is in the range of 0.01 mass% to 2 mass%, the predetermined amount of the stabilizing agent is in the range of 0.01 mass% to 3 mass%, the predetermined amount of the flavouring agent is in the range of 0 mass% to 1 mass% and the predetermined amount of the fluid medium is in the range of 75 mass% to 97 mass%, wherein the mass% of each ingredient is with respect to the total amount of the formulation.
In an exemplary embodiment of the present disclosure, the bacterial concentrated extract is present in an amount of 5.07 mass%, the natural repellent is present in an amount of 2.03 mass% the penetrating agent is present in an amount of 1.01 mass%, the surfactant is present in an amount of 0.203 mass%, the stabilizing agent is present in an amount of 0.304 mass%, the flavouring agent is present in an amount of 0.01 mass% and the fluid medium is present in an amount of 91.36 mass%, wherein the mass% of each ingredient is with respect to the total amount of the formulation.
In another exemplary embodiment of the present disclosure, the bacterial concentrated extract is present in an amount of 4.97 mass%, the natural repellent is present in an amount of 0.99 mass% the penetrating agent is present in an amount of 1.98 mass%, the surfactant is present in an amount of 0.248 mass%, the stabilizing agent is present in an amount of 0.223 mass%, the flavouring agent is present in an amount of 0.044 mass% and the fluid medium is present in an amount of 91.52 mass%, wherein the mass% of each ingredient is with respect to the total amount of the formulation.
In still another exemplary embodiment of the present disclosure, the bacterial concentrated extract is present in an amount of 5.02 mass%, the natural repellent is present in an amount of 1.50 mass% the penetrating agent is present in an amount of 1.50 mass%, the surfactant is present in an amount of 0.226 mass%, the stabilizing agent is present in an amount of 0.251 mass%, the flavouring agent is present in an amount of 0.031 mass% and the fluid medium is present in an amount of 91.44 mass%, wherein the mass% of each ingredient is with respect to the total amount of the formulation.
In yet another exemplary embodiment of the present disclosure, the bacterial concentrated extract is present in an amount of 5.07 mass%, the natural repellent is present in an amount of 1.77 mass% the penetrating agent is present in an amount of 1.26 mass%, the surfactant is present in an amount of 0.197 mass%, the stabilizing agent is present in an amount of 0.279 mass%, the flavouring agent is present in an amount of 0.045 mass% and the fluid medium is present in an amount of 91.36 mass%, wherein the mass% of each ingredient is with respect to the total amount of the formulation.
In still another exemplary embodiment of the present disclosure, the bacterial concentrated extract is present in an amount of 4.97 mass%, the natural repellent is present in an amount of 1.24 mass% the penetrating agent is present in an amount of 1.74 mass%, the surfactant is present in an amount of 0.198 mass%, the stabilizing agent is present in an amount of 0.298 mass%, the flavouring agent is present in an amount of 0.040 mass% and the fluid medium is present in an amount of 91.50 mass%, wherein the mass% of each ingredient is with respect to the total amount of the formulation.
In an embodiment of the present disclosure, the bacterial concentrated extract is obtained from at least one bacterial species selected from the group of Bacillus species and Priestia species.
The Bacillus species is selected from the group consisting of Bacillus velezensis, Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus aryabhattai and the Priestia species is Priestia megaterium.
In an embodiment of the present disclosure, the bacterial concentrated extract is extract of a combination of Bacillus velezensis and Priestia megaterium.
In an embodiment of the present disclosure, the bacterial concentrated extract is present in an amount in the range of 2 mass% to 10 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the bacterial concentrated extract is present in an amount of 5.07 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the bacterial concentrated extract is present in an amount of 4.97 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the bacterial concentrated extract is present in an amount of 5.02 mass% with respect to the total mass of the formulation.
In an exemplary embodiment of the present disclosure, a mass ratio of Bacillus velezensis extract to Priestia megaterium extract is in the range of 1:4 to 4:1 in the pesticidal formulation. In an exemplary embodiment of the present disclosure, a mass ratio of Bacillus velezensis extract to Priestia megaterium extract is 1:1 in the pesticidal formulation. In another exemplary embodiment of the present disclosure, a mass ratio of Bacillus velezensis extract to Priestia megaterium extract is 1:3 in the pesticidal formulation. In still another exemplary embodiment of the present disclosure, a mass ratio of Bacillus velezensis extract to Priestia megaterium extract is 3:1 in the pesticidal formulation.
The bacterial concentrated extract is an active ingredient of the pesticidal formulation.
In accordance with the present disclosure, the bacterial concentrated extract is obtained by culturing bacteria using suitable culture media. Further, the cultured bacterial cells undergo lysis and the bacterial metabolite concentrate from the lysate is extracted.
In an embodiment of the present disclosure, the natural repellent is selected from the group consisting of neem oil, castor oil, lemon grass oil and eucalyptus oil. In an exemplary embodiment of the present disclosure, the natural repellent is neem oil.
In an embodiment of the present disclosure, the predetermined amount of the natural repellent is in the range of 1 mass% to 5 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the predetermined amount of the natural repellent is 2.03 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the predetermined amount of the natural repellent is 0.99 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the predetermined amount of the natural repellent is 1.50 mass% with respect to the total mass of the formulation. In yet another exemplary embodiment of the present disclosure, the predetermined amount of the natural repellent is 1.77 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the predetermined amount of the natural repellent is 1.24 mass% with respect to the total mass of the formulation.
In an embodiment of the present disclosure, the penetrating agent is selected from the group consisting of isopropyl alcohol, ethanol and methanol. In an exemplary embodiment of the present disclosure, the penetrating agent is isopropyl alcohol.
In an embodiment of the present disclosure, the predetermined amount of the penetrating agent is in the range of 0.5 mass% to 5 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the predetermined amount of the penetrating agent is 1.01 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the predetermined amount of the penetrating agent is 1.98 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the predetermined amount of the penetrating agent is 1.50 mass% with respect to the total mass of the formulation. In yet another exemplary embodiment of the present disclosure, the predetermined amount of the penetrating agent is 1.26 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the predetermined amount of the penetrating agent is 1.74 mass% with respect to the total mass of the formulation.
In an embodiment of the present disclosure, the surfactant is selected from the group consisting of anionic surfactants, non-ionic surfactants, cationic surfactants and zwitterionic surfactants.
The anionic surfactant is selected from the group consisting of sodium laureth sulfate, sodium lauryl sarcosinate, sodium myreth sulfate, sodium pareth sulfate, sodium stearate, sodium lauryl sulfate, a-olefin sulfonate and ammonium laureth sulfate;
The non-ionic surfactant is selected from the group consisting of polyethylene glycol, glycerol, sorbitan monooleate (Span 80) and polysorbate 80 (tween 80).
The cationic surfactant is selected from the group consisting of benzethonium chloride, benzododecinium bromide ethyl lauroyl arginate, dimethyl dioctadecyl ammonium chloride and dioleoyl-3-trimethylammonium propane.
The zwitterionic surfactant is selected from the group consisting of betaines, sulfobetaines, and amine oxides.
In an exemplary embodiment of the present disclosure, the surfactant is ethylene oxide condensate of alkyl phenol (Alfox 200), which is the non-ionic surfactant.
In an embodiment of the present disclosure, the surfactant is present in an amount in the range of 0.01 mass% to 2 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the surfactant is present in an amount of 0.203 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the surfactant is present in an amount of 0.248 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the surfactant is present in an amount of 0.226 mass% with respect to the total mass of the formulation. In yet another exemplary embodiment of the present disclosure, the surfactant is present in an amount of 0.19 mass% with respect to the total mass of the formulation.
The surfactants act as adjuvants in the pesticidal formulation which effectively enhances the biological efficacy by promoting penetration and spread.
In an embodiment of the present disclosure, the stabilizing agent is selected from the group consisting of sodium carbonate, potassium bicarbonate and sodium bicarbonate. In an exemplary embodiment of the present disclosure, the stabilizing agent is sodium carbonate.
The stabilizing agent stabilizes the formulation.
In an embodiment of the present disclosure, the stabilizing agent is present in an amount in the range of 0.01 mass% to 3 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the stabilizing agent is present in an amount of 0.304 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the stabilizing agent is present in an amount of 0.223 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the stabilizing agent is present in an amount of 0.251 mass% with respect to the total mass of the formulation. In yet another exemplary embodiment of the present disclosure, the stabilizing agent is present in an amount of 0.279 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the stabilizing agent is present in an amount of 0.298 mass% with respect to the total mass of the formulation.
In an embodiment of the present disclosure, the flavouring agent is selected from the group consisting of lemon grass oil and eucalyptus oil. In an exemplary embodiment of the present disclosure, the flavouring agent is lemon grass oil.
In an embodiment of the present disclosure, the flavouring agent is present in an amount in the range of 0 mass% to 1 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the flavouring agent is present in an amount of 0.01 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the flavouring agent is present in an amount of 0.04 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the flavouring agent is present in an amount of 0.03 mass% with respect to the total mass of the formulation.
In an embodiment of the present disclosure, the fluid medium is water.
In an embodiment of the present disclosure, the fluid medium is present in an amount in the range of 75 mass% to 97 mass% with respect to the total mass of the pesticidal formulation. In an exemplary embodiment of the present disclosure, the fluid medium is present in an amount of 91.36 mass% with respect to the total mass of the pesticidal formulation. In another exemplary embodiment of the present disclosure, the fluid medium is present in an amount of 91.52 mass% with respect to the total mass of the pesticidal formulation. In still another exemplary embodiment of the present disclosure, the fluid medium is present in an amount of 91.44 mass% with respect to the total mass of the pesticidal formulation. In yet another exemplary embodiment of the present disclosure, the fluid medium is present in an amount of 91.36 mass% with respect to the total mass of the pesticidal formulation. In still another exemplary embodiment of the present disclosure, the fluid medium is present in an amount of 91.50 mass% with respect to the total mass of the pesticidal formulation.
In accordance with the present disclosure, the pesticidal formulation exhibits 100% solubility in water with remarkable ratio of 1 ml in 100 ml (1: 100 solubility).
In an embodiment of the present disclosure, the pesticidal formulation is in the form of a liquid.
The pesticidal formulation of the present disclosure can be used against pests of family Pseudococcidae, particularly, for mealy bugs.
In another aspect, the present disclosure provides a process for the preparation of a pesticidal formulation.
In an embodiment of the present disclosure, the process for the preparation of the pesticidal formulation comprises mixing predetermined amounts of at least one bacterial concentrated extract, a natural repellent, a penetrating agent, a surfactant, a stabilizing agent, optionally a flavouring agent in a predetermined amount of a fluid medium under stirring for a predetermined time at a predetermined temperature to obtain the pesticidal formulation.
In an embodiment of the present disclosure, the bacterial concentrated extract is obtained from at least one bacterial species selected from the group consisting of Bacillus species and Priestia species.
The Bacillus species is selected from the group consisting of Bacillus velezensis, Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus aryabhattai and the Priestia species is Priestia megaterium.
In an embodiment of the present disclosure, the bacterial concentrated extract is extract of a combination of Bacillus velezensis and Priestia megaterium.
In an exemplary embodiment of the present disclosure, a mass ratio of Bacillus velezensis extract to Priestia megaterium extract is in the range of 1:4 to 4:1 in the pesticidal formulation. In an exemplary embodiment of the present disclosure, a mass ratio of Bacillus velezensis extract to Priestia megaterium extract is 1:1 in the pesticidal formulation. In another exemplary embodiment of the present disclosure, a mass ratio of Bacillus velezensis extract to Priestia megaterium extract is 1:3 in the pesticidal formulation. In still another exemplary embodiment of the present disclosure, a mass ratio of Bacillus velezensis extract to Priestia megaterium extract is 3:1 in the pesticidal formulation.
In accordance with the present disclosure, the bacterial concentrated extract is obtained by culturing bacteria using suitable culture media. Further, the cultured bacterial cells present in broth are lysed by using a fluid medium to obtain a mixture comprising lysed bacterial cells and the bacterial metabolites. The mixture is concentrated to obtain the bacterial concentrated extract.
In an embodiment of the present disclosure, the predetermined amount of the bacterial concentrated extract is in the range of 2 mass% to 10 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the bacterial concentrated extract is present in an amount of 5.07 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the bacterial concentrated extract is present in an amount of 4.97 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the bacterial concentrated extract is present in an amount of 5.02 mass% with respect to the total mass of the formulation.
In an embodiment of the present disclosure, the natural repellent is selected from the group consisting of neem oil, castor oil, lemon grass oil and eucalyptus oil. In an exemplary embodiment of the present disclosure, the natural repellent is neem oil.
In an embodiment of the present disclosure, the predetermined amount of the natural repellent is in the range of 1 mass% to 5 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the predetermined amount of the natural repellent is 2.03 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the predetermined amount of the natural repellent is 0.99 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the predetermined amount of the natural repellent is 1.50 mass% with respect to the total mass of the formulation. In yet another exemplary embodiment of the present disclosure, the predetermined amount of the natural repellent is 1.77 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the predetermined amount of the natural repellent is 1.24 mass% with respect to the total mass of the formulation.
In an embodiment of the present disclosure, the penetrating agent is selected from the group consisting of isopropyl alcohol, ethanol and methanol. In an exemplary embodiment of the present disclosure, the penetrating agent is isopropyl alcohol.
In an embodiment of the present disclosure, the predetermined amount of the penetrating agent is in the range of 0.5 mass% to 5 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the predetermined amount of the penetrating agent is 1.01 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the predetermined amount of the penetrating agent is 1.98 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the predetermined amount of the penetrating agent is 1.50 mass% with respect to the total mass of the formulation. In yet another exemplary embodiment of the present disclosure, the predetermined amount of the penetrating agent is 1.26 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the predetermined amount of the penetrating agent is 1.74 mass% with respect to the total mass of the formulation.
In an embodiment of the present disclosure, the surfactant is selected from the group consisting of anionic surfactants, non-ionic surfactants, cationic surfactants and zwitterionic surfactants.
The anionic surfactant is selected from the group consisting of sodium laureth sulfate, sodium lauryl sarcosinate, sodium myreth sulfate, sodium pareth sulfate, sodium stearate, sodium lauryl sulfate, a-olefin sulfonate and ammonium laureth sulfate.
The non-ionic surfactant is selected from the group consisting of polyethylene glycol, glycerol, sorbitan monooleate (Span 80), ethylene oxide condensate of alkyl phenol (Alfox 200) and polysorbate 80 (tween 80).
The cationic surfactant is selected from the group consisting of benzethonium chloride, benzododecinium bromide ethyl lauroyl arginate, dimethyl dioctadecyl ammonium chloride and dioleoyl-3-trimethylammonium propane.
The zwitterionic surfactant is selected from the group consisting of betaines, sulfobetaines and amine oxides.
In an exemplary embodiment of the present disclosure, the surfactant is ethylene oxide condensate of alkyl phenol (Alfox 200), which is the non-ionic surfactant.
In an embodiment of the present disclosure, the predetermined amount of the surfactant is in the range of 0.01 mass% to 2 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the surfactant is present in an amount of 0.203 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the surfactant is present in an amount of 0.248 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the surfactant is present in an amount of 0.226 mass% with respect to the total mass of the formulation. In yet another exemplary embodiment of the present disclosure, the surfactant is present in an amount of 0.19 mass% with respect to the total mass of the formulation.
In an embodiment of the present disclosure, the stabilizing agent is selected from the group consisting of sodium carbonate, potassium bicarbonate and sodium bicarbonate. In an exemplary embodiment of the present disclosure, the stabilizing agent is sodium carbonate.
In an embodiment of the present disclosure, the predetermined amount of the stabilizing agent is in the range of 0.01 mass% to 3 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the stabilizing agent is present in an amount of 0.304 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the stabilizing agent is present in an amount of 0.223 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the stabilizing agent is present in an amount of 0.251 mass% with respect to the total mass of the formulation. In yet another exemplary embodiment of the present disclosure, the stabilizing agent is present in an amount of 0.279 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the stabilizing agent is present in an amount of 0.298 mass% with respect to the total mass of the formulation.
In an embodiment of the present disclosure, the flavouring agent is selected from the group consisting of lemon grass oil and eucalyptus oil. In an exemplary embodiment of the present disclosure, the flavouring agent is lemon grass oil.
In an embodiment of the present disclosure, the predetermined amount of the flavouring agent is in the range of 0 mass% to 1 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the flavouring agent is present in an amount of 0.01 mass% with respect to the total mass of the formulation. In another exemplary embodiment of the present disclosure, the flavouring agent is present in an amount of 0.04 mass% with respect to the total mass of the formulation. In still another exemplary embodiment of the present disclosure, the flavouring agent is present in an amount of 0.03 mass% with respect to the total mass of the formulation.
In an embodiment of the present disclosure, the fluid medium is water.
In an embodiment of the present disclosure, the predetermined amount of the fluid medium is in the range of 75 mass% to 97 mass% with respect to the total mass of the formulation. In an exemplary embodiment of the present disclosure, the fluid medium is present in an amount of 91.36 mass% with respect to the total mass of the pesticidal formulation. In another exemplary embodiment of the present disclosure, the fluid medium is present in an amount of 91.52 mass% with respect to the total mass of the pesticidal formulation. In still another exemplary embodiment of the present disclosure, the fluid medium is present in an amount of 91.44 mass% with respect to the total mass of the pesticidal formulation. In yet another exemplary embodiment of the present disclosure, the fluid medium is present in an amount of 91.36 mass% with respect to the total mass of the pesticidal formulation. In still another exemplary embodiment of the present disclosure, the fluid medium is present in an amount of 91.50 mass% with respect to the total mass of the pesticidal formulation.
The pesticidal formulation prepared in accordance with the present disclosure can be used against pests of family Pseudococcidae, particularly, for mealy bugs.
In an embodiment of the present disclosure, the predetermined time period is in the range of 15 minutes to 60 minutes. In an exemplary embodiment of the present disclosure, the predetermined time period is 30 minutes.
In an embodiment of the present disclosure, the predetermined temperature is in the range of 25 ºC to 35 ºC. In an exemplary embodiment of the present disclosure, the predetermined temperature is 30 ºC.
The process for the preparation of the pesticidal formulation in accordance with the present disclosure is simple, economic, environment friendly and is convenient for industrial scale-up.
The pesticidal formulation of the present disclosure can be used against the pests of family Pseudococcidae. The pesticidal formulation of the present disclosure is ready to use (RTU) type formulation.
The foregoing description of the embodiments has been provided for purposes of illustration and not intended to limit the scope of the present disclosure. Individual components of a particular embodiment are generally not limited to that particular embodiment but are interchangeable. Such variations are not to be regarded as a departure from the present disclosure, and all such modifications are considered to be within the scope of the present disclosure.
The present disclosure is further described in light of the following experiments which are set forth for illustration purposes only and not to be construed for limiting the scope of the disclosure. The following experiments can be scaled up to industrial/commercial scale and the results obtained can be extrapolated to industrial scale.
EXPERIMENTAL DETAILS:
Experiment 1: Process for the preparation of a pesticidal formulation in accordance with the present disclosure
General procedure: Predetermined amounts of at least one bacterial concentrated extract, a natural repellent, a penetrating agent, a surfactant, a stabilizing agent, optionally a flavouring agent were mixed in a predetermined amount of a fluid medium under stirring at a 30ºC for 30 minutes to obtain the pesticidal formulation.
The so obtained pesticidal formulation was directly applied on the infected plants.
The pesticidal formulations were prepared in Examples E1 to E5 by following the general procedure as mentioned above. The predetermined amounts of the specific ingredients of the pesticidal formulations are as given in Table 1.
Table 1: Predetermined amounts of the ingredients used in examples E1-E5
Ingredients and their function E1
(amount in Kg) E2
(amount in Kg) E3
(amount on Kg) E4
(amount in Kg) E5
(amount in kg)
Sr. no. Ingredient Function
1. Bacillus velezensis Bacterial concentrated extract 25 Kg 12.5 37.5 35 15
Priestia megaterium Bacterial concentrated extract 25 Kg 37.5 12.5 15 35
2. Neem oil Natural repellent 20 Kg 10 15 17.5 12.5
3. Isopropyl alcohol (IPA) Penetrating agent 10 Kg 20 15 12.5 17.5
4. Alfox 200 Surfactant 2 Kg 2.5 2.25 1.95 2.0
5. Sodium carbonate (Na2CO3) Stabilizing agent 3 Kg 2.25 2.5 2.75 3
6. Lemon grass oil Flavouring agent 100 ml 500 350 500 450
7. Water Fluid medium 900 Kg 920 910 900 920
For the sake of brevity, multiple number of experiments were not included in the specification. However, other ingredients having similar functionality will give similar results for the pesticidal formulation.
(A) Evaluation of insecticidal efficacy of the pesticidal formulation in accordance with the present disclosure
For the evaluation of insecticidal efficacy Abbott's formula was considered. This formula serves as a quantitative tool for precisely gauging the effectiveness of the product under controlled laboratory conditions.
Pesticidal efficacy was calculated by the following formula
Percentage of Population Reduction = T1 - T2/T1 × 100
Where,
T1= Number of populations before treatment in the treated sample.
T2= Number of Population after treatment in the treated sample.
(I) Evaluation of efficacy at field level:
The efficacy of the pesticidal formulation was studied in the fields of Gangasandra village premises in Karnataka. The pesticidal formulation prepared in Experiment 1 was sprayed on the infected plant leaves of brinjal plants. Additionally, water was employed as a negative control, while commercially available pesticidal formulation (lazy Gardener) was utilized as the positive control. These pesticidal formulations and controls were then subjected to rigorous efficacy studies carried out at the field level and pesticide efficacy was calculated according to Abbot’s formula. The results of the efficacy study at field level are tabulated in Table 2 below.
Table 2: Population data of mealy bugs at field level

Sl. No.
Sample Total Population (In Number*)
% of Population reduction
Before treatment After treatment
1 Negative Control (Water) 45 42 6.66%
2 Pesticidal formulation of Experiment 1 45 5 88.88%
3 Positive control Commercially available pesticidal formulation (Lazy Gardener) 45 25 44.44%
* The mealy bugs were counted by direct counting method wherein the mealy bugs infestation was measured by counting the insects inside a selected region.
From Table 2, it was observed that the pesticidal formulation of the present disclosure showed significant control of mealy bugs and maximum % population reduction for mealy bugs as compared to the commercially available pesticidal formulation (Lazy gardener) and negative control (water). Further, with the use of the pesticidal formulation of the present disclosure, the mealy bugs turned to dark brown color, indicating mortality of the mealy bugs.
Figure 1 illustrates (A) images of mealy bugs infested plant leaves before treatment with the commercially available pesticidal formulation (Lazy gardener), (B) images of mealy bugs infested plant leaves before treatment with the pesticidal formulation in accordance with the present disclosure, (C) images of mealy bugs infested plant leaves after treatment with the commercially available pesticidal formulation, (D) images of mealy bugs infested plant leaves after treatment with the pesticidal formulation in accordance with the present disclosure, (E) microscopic images of mealy bugs infested plant leaves after treatment with the commercially available pesticidal formulation (Lazy gardener), (F) microscopic images of mealy bugs infested plant leaves after treatment with the pesticidal formulation in accordance with the present disclosure.
(B) Phytotoxicity study of the pesticidal formulation in accordance with the present disclosure
The pesticidal formulation as prepared in Experiment 1 was tested for phytotoxicity on brinjal plants.
The results are evident from Figure 1(B) and (D), wherein Figure 1(B) shows the image of mealy bugs infested plant leaves after treatment with the commercially available pesticidal formulation and Figure 1(D) shows the images of mealy bugs infested plant leaves after treatment with a pesticidal formulation of the present disclosure.
It was observed that the brinjal plants treated with the pesticidal formulation of the present disclosure (ready to use) showed no phytotoxic symptoms such as there is no leaf injury observed on tips and leaf surfaces. Further, there is no wilting, necrosis, vein clearing, epinasty and hyponasty were observed in the leaves.
Stability study for the pesticidal formulation was also conducted.
The process involved the preparation of the pesticidal formulation followed by subjecting it to rigorous stability assessments at various temperature conditions. The selected temperatures encompassed a range of environmental and physiological scenarios, aiming to gauge the product's (pesticidal formulation of the present disclosure) resilience and durability. Throughout a continuous 15-days observation period under controlled conditions, the pesticidal formulation of the present disclosure underwent stability evaluation without any external interventions. The pesticidal formulation was found to be stable. Upon completion of 15-days acceleration period, the same set of samples that were subjected to the stability studies were selected for a comprehensive quality assessment.
Following Table 3 enlists the various parameters studied for the pesticidal formulation in accordance with the present disclosure.
Table 3: Various parameters testing of the pesticidal formulation in accordance with the present disclosure
Sr. No. Parameters Test Method Reference range Observed range
1. Appearance Visual method Creamy white colored liquid Creamy white colored liquid
2. pH pH meter method 8.5 to 10.5 9.87
3. Specific gravity Specific gravity bottle method 0.98 to 1.01 0.99
4. Free fatty acids % by wt (%) Titration method Nil Nil
5. Crude protein % by weight (%) Kjeldahl method Nil Nil
6. Total nitrogen % by weight (%) Kjeldahl method Nil Nil
7. Amino acids % by weight (%) Soromen’s method Nil Nil
8. Carbohydrates % by weight Anthrone reagent method Nil Nil
9. Solubility --- Soluble in water Soluble in water
10. Odour --- Characteristic odor Characteristic odor
From Table 3, it was observed that all the values of the parameters of the pesticidal formulation of the present disclosure are within the range of reference range and therefore, the pesticidal formulation of the present disclosure is effective to treat against mealy bugs.
TECHNICAL ADVANCEMENTS
The present disclosure described herein above has several technical advantages including, but not limited to, the realization of;
? a pesticidal formulation that:
• is effective against mealy bugs; and
• is non-phytotoxic even at high dosage; and
? a process for the preparation of a pesticidal formulation that is
• simple, economic and environment friendly.
The embodiments herein and the various features and advantageous details thereof are explained with reference to the non-limiting embodiments in the following description. Descriptions of well-known components and processing techniques are omitted so as to not unnecessarily obscure the embodiments herein. The examples used herein are intended merely to facilitate an understanding of ways in which the embodiments herein may be practiced and to further enable those of skill in the art to practice the embodiments herein. Accordingly, the examples should not be construed as limiting the scope of the embodiments herein.
The foregoing description of the specific embodiments so fully reveal the general nature of the embodiments herein that others can, by applying current knowledge, readily modify and/or adapt for various applications such specific embodiments without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments. It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. Therefore, while the embodiments herein have been described in terms of preferred embodiments, those skilled in the art will recognize that the embodiments herein can be practiced with modification within the spirit and scope of the embodiments as described herein.
The use of the expression “at least” or “at least one” suggests the use of one or more elements or ingredients or quantities, as the use may be in the embodiment of the disclosure to achieve one or more of the desired object or results.
Any discussion of documents, acts, materials, devices, articles or the like that has been included in this specification is solely for the purpose of providing a context for the disclosure. It is not to be taken as an admission that any or all of these matters form a part of the prior art base or were common general knowledge in the field relevant to the disclosure as it existed anywhere before the priority date of this application.
The numerical values mentioned for the various physical parameters, dimensions or quantities are only approximations and it is envisaged that the values higher/lower than the numerical values assigned to the parameters, dimensions or quantities fall within the scope of the disclosure, unless there is a statement in the specification specific to the contrary.
While considerable emphasis has been placed herein on the components and component parts of the preferred embodiments, it will be appreciated that many embodiments can be made and that many changes can be made in the preferred embodiments without departing from the principles of the disclosure. These and other changes in the preferred embodiment as well as other embodiments of the disclosure will be apparent to those skilled in the art from the disclosure herein, whereby it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustrative of the disclosure and not as a limitation. ,CLAIMS:WE CLAIM:
1. A pesticidal formulation comprises:
a. a predetermined amount of at least one bacterial concentrated extract;
b. a predetermined amount of a natural repellent;
c. a predetermined amount of a penetrating agent;
d. a predetermined amount of a surfactant;
e. a predetermined amount of a stabilizing agent;
f. optionally a predetermined amount of a flavouring agent; and
g. a predetermined amount of a fluid medium.
2. The formulation as claimed in claim 1, wherein
• said predetermined amount of said bacterial concentrated extract is in the range of 2 mass% to 10 mass%;
• said predetermined amount of said natural repellent is in the range of 1 mass% to 5 mass%;
• said predetermined amount of said penetrating agent is in the range of 0.5 mass% to 5 mass%;
• said predetermined amount of said surfactant is in the range of 0.01 mass% to 2 mass%;
• said predetermined amount of said stabilizing agent is in the range of 0.01 mass% to 3 mass%;
• said predetermined amount of said flavouring agent is in the range of 0 mass% to 1 mass%; and
• said predetermined amount of said fluid medium is in the range of 75 mass% to 97 mass%,
wherein said mass% of each ingredient is with respect to the total amount of said formulation.
3. The formulation as claimed in claim 1, wherein said bacterial concentrated extract is obtained from at least one bacterial species selected from the group consisting of Bacillus species and Priestia species.
4. The formulation as claimed in claim 3, wherein
• said Bacillus species is selected from the group consisting of Bacillus velezensis, Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus aryabhattai; and
• said Priestia species is Priestia megaterium.
5. The formulation as claimed in claim 3, wherein said bacterial concentrated extract is extract of a combination of Bacillus velezensis and Priestia megaterium.
6. The formulation as claimed in claim 1, wherein
• said natural repellent is selected from the group consisting of neem oil, castor oil, lemon grass oil and eucalyptus oil;
• said penetrating agent is selected from the group consisting of isopropyl alcohol, ethanol and methanol;
• said surfactant is selected from the group consisting of anionic surfactants, non-ionic surfactants, cationic surfactants and zwitterionic surfactants;
• said stabilizing agent is selected from the group consisting of sodium carbonate, potassium bicarbonate and sodium bicarbonate;
• said flavouring agent is selected from the group consisting of lemon grass oil and eucalyptus oil; and
• said fluid medium is water.
7. The formulation as claimed in claim 6, wherein
• said anionic surfactant is selected from the group consisting of sodium laureth sulfate, sodium lauryl sarcosinate, sodium myreth sulfate, sodium pareth sulfate, sodium stearate, sodium lauryl sulfate, a-olefin sulfonate and ammonium laureth sulfate;
• said non-ionic surfactant is selected from the group consisting of polyethylene glycol, glycerol, sorbitan monooleate, ethylene oxide condensate of alkyl phenol and polysorbate 80;
• said cationic surfactant is selected from the group consisting of benzethonium chloride, benzododecinium bromide ethyl lauroyl arginate, dimethyl dioctadecyl ammonium chloride and dioleoyl-3-trimethylammonium propane; and
• said zwitterionic surfactant is selected from the group consisting of betaines, sulfobetaines, and amine oxides.
8. The formulation as claimed in claim 1 is in the form of a liquid.
9. A process for the preparation of a pesticidal formulation as claimed in claim 1, wherein said process comprises mixing predetermined amounts of at least one bacterial concentrated extract, a natural repellent, a penetrating agent, a surfactant, a stabilizing agent, optionally a flavouring agent in a predetermined amount of a fluid medium under stirring for a predetermined time period at a predetermined temperature to obtain said pesticidal formulation.
10. The process as claimed in claim 9, wherein
• said predetermined time period is in the range of 15 minutes to 60 minutes; and
• said predetermined temperature is in the range of 25 oC to 35 oC.
11. The process as claimed in claim 9, wherein
• said natural repellent is selected from the group consisting of neem oil, castor oil, lemon grass oil and eucalyptus oil;
• said penetrating agent is selected from the group consisting of isopropyl alcohol, ethanol and methanol;
• said surfactant is selected from the group consisting of anionic surfactants, non-ionic surfactants, cationic surfactants and zwitterionic surfactants;
• said stabilizing agent is selected from the group consisting of sodium carbonate, potassium bicarbonate and sodium bicarbonate;
• said flavouring agent is selected from the group consisting of lemon grass oil and eucalyptus oil; and
• said fluid medium is water.
12. The process as claimed in claim 11, wherein
• said anionic surfactant is selected from the group consisting of sodium laureth sulfate, sodium lauryl sarcosinate, sodium myreth sulfate, sodium pareth sulfate, sodium stearate, sodium lauryl sulfate, a-olefin sulfonate and ammonium laureth sulfate;
• said non-ionic surfactant is selected from the group consisting of polyethylene glycol, glycerol, sorbitan monooleate, ethylene oxide condensate of alkyl phenol and polysorbate 80;
• said cationic surfactant is selected from the group consisting of benzethonium chloride, benzododecinium bromide ethyl lauroyl arginate, dimethyl dioctadecyl ammonium chloride and dioleoyl-3-trimethylammonium propane; and
• said zwitterionic surfactant is selected from the group consisting of betaines, sulfobetaines, and amine oxides.
13. The process as claimed in claim 9, wherein
• said predetermined amount of said bacterial concentrated extract is in the range of 2 mass% to 10 mass%;
• said predetermined amount of said natural repellent is in the range of 1 mass% to 5 mass%;
• said predetermined amount of said penetrating agent is in the range of 0.5 mass% to 5 mass%;
• said predetermined amount of said surfactant is in the range of 0.01 mass% to 2 mass%;
• said predetermined amount of said stabilizing agent is in the range of 0.01 mass% to 3 mass%;
• said predetermined amount of said flavouring agent is in the range of 0 mass% to 1 mass%; and
• said predetermined amount of said fluid medium is in the range of 75 mass% to 97 mass%,
wherein said mass% of each ingredient is with respect to the total amount of said formulation.
14. The pesticidal formulation as claimed in claim 1 is used against pests of family Pseudococcidae.

Dated this 2nd day of May, 2024

_______________________________
MOHAN RAJKUMAR DEWAN, IN/PA – 25
OF R. K. DEWAN & CO.
AUTHORIZED AGENT OF APPLICANT

TO,
THE CONTROLLER OF PATENTS
THE PATENT OFFICE, AT CHENNAI