TECHNICAL FIELD OF THE INVENTION

The present invention relates generally to the pesticide composition and more particularly to a synergistic plant-based composition mainly used to control insects, acarus, aphids, termites, nematodes, viruses and bacteria in agriculture field and domestic and civil disinfestation. The present invention specifically relates to a composition which based on natural extracts.

BACKGROUND OF THE INVENTION

Currently various chemicals and chemical compositions have been in use at great extent in the pesticides, which are being used to control the insects, termites, nematodes and so forth. Further several natural compositions are also in use to achieve the same goal, which show less or no toxicity to the consumers who consume it, though they can have severe impact on the environment as they are not easily eliminated. The compositions can specifically target the insects, termites, nematodes and so forth, but can pose as a toxic to non-target species.

Plant parasitic nematodes cause significant damage to a wide variety of crops, resulting in global crop yield losses estimated to range from 5% to 12% annually. Root damage by nematodes is very common and leads to stunted plants, which have smaller root systems, show symptoms of mineral deficiencies in their leaves and wilt easily. Damage by nematodes also predisposes plants to infection by a wide variety of plant pathogenic fungi and bacteria.

In order to combat and control nematodes, farmers typically use chemical nematicides. These range from gas and liquid fumigation, such as methyl bromide and chloropicrin, to application of organophosphates and carbamates, such as thionazin and oxamyl. Use of these chemical nematicides has been ongoing for several decades. Despite the effectiveness of the chemical nematicide in controlling target nematodes, there are serious limitations to these methods. One limitation is that chemical nematicides cannot act against nematodes that have already penetrated the root. Another limitation is the danger associated with the production and use of chemical nematicides. Chemical nematicides are highly toxic and can lead to human poisoning and death. As a result, countries have restricted and sometimes banned certain pesticides. Methyl bromide in particular is banned in most countries due to its ozone depleting effects.

Because of these restrictions and bans, there is a lack of viable nematode solutions. The present invention provides a safe and effective means to replace or lessen the use of chemical pesticides. It is also unique in providing a solution that both inhibits nematode penetration into the plant root and then prevents maturation of those nematodes that manage to overcome this initial barrier.

Plants are constantly challenged by a wide variety of pathogenic organisms including viruses, bacteria, fungi, and nematodes. Attempts have been made to control plant disease by means of disinfections, replacement of the soil, various cultural practices, and control by chemicals. Some plants suffer from detrimental soil-spread diseases, which have not been possible to control owing to restrictions of use of chemical control agents and hazard periods due to possible residues or lack of sufficiently effective products.

The control of fungi is important since fungal growth on plants or on parts of plants inhibits production of foliage, fruit or seed, and the overall quality of a cultivated crop. About twenty-five (25) percent of all fungal diseases in agricultural and horticulture are caused by powdery mildew phytopathogens.

Due to the vast economic ramifications of fungal propagation in agricultural and horticultural cultivations, a broad spectrum of fungicidal and fungi static products has been developed for general and specific applications. Such examples are the use of inorganic bicarbonate, carbonate compounds, lecithin, and lime. However, these fungicidal and fungistatic products may be harmful to the environment and may pollute areas such as ground waters. Thus, there is a need for an organic fungal treatment which provides for an inexpensive way to control fungi without harmingthe environment and protect the plants with a minimum of phytotoxic side effects.

Since some bacteria, molds, yeasts and viruses can lead to serious illnesses, disinfection from the everyday life of medicine and private household is indispensable. The importance of disinfection for people's well-being is often underestimated. Thus, in earlier centuries, more people died of the great epidemics (plague, cholera, smallpox or flu) than were killed in wars. Until the beginning of the 20th century, severe bacterial infections were often deadly diseases in industrialized countries. In third world countries, even today, infectious diseases, which are mostly due to inadequate hygienic conditions, are causing countless casualties. Therefore, especially in the medical field, there is still a high demand for effective and inexpensive disinfectants.

There are several prior art patents which provides different compositions to control the insects, termites, nematodes and so forth.

WIPO patent application number 2015043986A1 by Chemia S.P.A. discloses a pesticide based on natural extracts including Thymol, an alkaloid Matrine, Carvacrol and at least one molecule chosen from Ossimatrine, Sophocarpine, N- oxysophocarpine, Cytisine, Aloperine, Carangine and Toosedanin. Said pesticide is particularly suitable for use in agriculture, gardening, hobbies, veterinary, for wood protection, domestic and civil environment, open or closed public places against theharmful effects of insects, mites, aphids, nematodes and virus diseases of plants.

US patent application 8501247B2 by Tyratech Inc, Mondelez Global LLC discloses pest control using natural pest control agent blends for controlling a target pest, wherein the composition includes at least two active ingredients selected from the group consisting of thymyl acetate, linalyl acetate, amyl butyrate, anise star oil, black seed oil, p-cymene, geraniol, isopropyl myristate, d-limonene, linalool, lilac flower oil, methyl salicylate, alpha-pinene, piperonal, piperonyl alcohol, tetrahydrolinalool, thyme oil white, thyme oil red, thymol, vanillin, and winter- green oil, wherein the composition causes synergistic control of the target pest.

US patent 20060194698A by University of Tennessee Research Foundation discloses the use of herbs as a delivery system for bioactive phytochemicals for controlling weeds, plant pests, or plant pathogens comprising the application of a bioactive herbage (plant material) composition to soil as a soil amendment or as top dressing for potted plants. In certain embodiments, the method of controlling weeds, plant pests, or plant pathogens comprises the application of a bioactive herbage (plant material) composition to soil as a soil amendment or as top dressing for potted plants in amounts sufficient to control weeds, plant pests, or plant pathogens, wherein said bioactive herbage is obtained from: (a) Monarda spp.;(b) Chamaemelum spp.; (c) Matricaria spp.; (d) Chenopodium spp.; or (e) various combinations of thereof. Herbage can be dried or wet and other sources of bioactive herbage are also suitable for use in the subject invention is provided.

WIPO patent application 2006097700A1 by Plant Impact Pic discloses plant growth enhancement and method thereof wherein said method comprising applying to a plant or to the environment thereof, an agricultural composition comprising (i) one or more essential oils, or active components thereof, or mixtures of these, (ii) a plant oil and (iii) an emulsifier. Such compositions enhance plant growth, in particular by enhancing nutrient uptake from the soil or the environment. Compositions for use in the method are also described and claimed.

Indian patent application number 245/MUM/2004 by North Maharashtra University discloses A process for preparing a botanical pesticide from seeds of Annona Squamosa, characterized in that (i) defatting seed powder of A. squamosa with defatting solvent(s); (ii) treating the residue cake obtained in step (i) with a polar solvent; (iii) filtering the extract of step (ii); (iv) concentrating the filtered extract of step (iii); (v) partitioning the concentrated extract of step (iv) with a suitable extracting solvent; (vi) adding suitable dehydrating agent in extract of step (v); (vii) completely drying the extract of step (vi); (viii) dissolving the residue of step (vii) in a suitable extracting solvent; (ix) further defatting extract of step (viii) with defatting solvent; (x) drying completely/partially the extract of step (ix); (xi) dissolving the residue of step (x) in a suitable polar solvent and (xii) purifying by passing through a column as described herein before.

In order to achieve a strong disinfecting effect, in the past, above all, highly persistent chemical compounds have been used as disinfectants, which preserve their disinfecting effect as long as possible and thus lead to an effective and long- lasting protection against microorganisms. However, this leads to major environmental problems. The highly persistent disinfectants accumulate in the groundwater or in the food chain and lead to ecological and health problems. For example, environmental problems can arise when disinfectants reach biological sewage treatment plants in high concentrations.

In particular, the disinfecting compositions according to the invention should exhibit the lowest possible toxicity, be ecologically compatible, i. show no accumulation in groundwater and food chain. Despite these properties, the compositions according to the invention should nevertheless have strong and long- lasting disinfecting effects and thus offer reliable protection against microorganisms

OBJECTS OF THE INVENTION:

The primary object of the present invention is to provide a composition to control plant pathogenic fungi and parasitic nematodes.

Another object of the present invention is to provide a composition of plant extracts which is organic alternative to the chemical-based pesticides.

Yet another object of the present invention is to provide a plant based composition comprising of extract of Thymus Vulgaris containing Monoterpenoid phenol derivatives of p-cymene isomeric with carvacrol polyphenols as an anti-fungal agent, Extract of Annona Squamosa containing acetogenins from class of polyketide containing 32 or 34 carbon chain having oxygenated functional group including Hydroxyls, Ketones, Expoxides, Tetrahydrofuran and Tetrahydropyrans as insecticidal agent and 5-Chloro-2-(4-chlorophenoxy) phenol as anti bacterial, anti-viral agent.

SUMMARY OF THE INVENTION

The present invention discloses a natural synergistic composition of plant extracts which acts anti-fungal, anti-bacterial and insecticidal composition. The said synergistic composition comprises of plant-based extracts wherein the various forms of the said composition comprising of Organic or aqueous extract of Thymus vulgaris as anti-fungal component, Organic or aqueous extract of Annona Squamosa as insecticidal component, 5-Chloro-2-(4- chlorophenoxy) phenol asanti-bacterial component.

Surfactants preferably selected from the group of Naturally Ethoxylated/Propoxylated or Ethoxylated-Propoxylated Natural Vegetable oil including but not limited to castor oil, Palm Oil, Coconut Oil, Soybean oil as natural surfactant. However, any other surfactant which is having HLB (Hydrophile Lypophile Balance) of 9 to 20 can be used in the formulation described in the present invention.

Diluents preferably selected from the group of but not limited to Natural oil and its derivatives from the sources like soyabean, cotton or palm, and 2-ethyl hexyl lactate/dimethyl sulfoxide/water/N, N Dimethyl decanamide and water.

According to the present invention, the said plant-based composition is highly effective, natural and low toxic composition, which can be used to control plant pathogenic fungi, plant nematodes, viruses and bacteria.

DETAILED DESCRIPTION OF INVENTION

Aspects of the present disclosure relates to a composition based on the natural extracts to eliminate and control the plant nematodes and to more particularly to a synergistic plant- based composition mainly used to control insects, acarus, aphids, termites, nematodes, Bacteria and viruses in agriculture field and domestic and civildisinfestation.

In the following description, numerous specific details are set forth in order to provide a thorough understanding of embodiments of the present invention. It will be apparent to one skilled in the art that embodiments of the present invention may be practiced without some of these specific details.

Detailed embodiments of the present invention are disclosed herein below. However, it is to be understood that the disclosed embodiments are merely exemplary of the invention, which can be embodied in various forms. The scope of the invention is not limited to the disclosed embodiments and terms and phrases used herein are not intended to be limiting but rather to provide an understandabledescription of the invention.

If the specification states a component or feature “may”, “can”, “could”, or “might” be included or have a characteristic, that particular component or feature is not required to be included or have the characteristic.

The present invention discloses a natural synergistic comprises of plant-based extracts wherein the various forms of the said composition comprising of Organic or aqueous extract of Thymus vulgaris as anti-fungal component, Organic or aqueous extract of Annona Squamosa as insecticidal component, 5-Chloro-2-(4- chlorophenoxy) phenol as anti-bacterial component.

The synergistic composition comprises of plant-based extracts wherein the various forms of the said composition comprising of Organic or aqueous extract of Thymus vulgaris as anti-fungal component, which acts against the fungal infections seen in the plants. Thymus Vulgaris has antibacterial, antifungal and antivirus properties. The antifungal potential of Thymus Vulgaris , it can be used in agriculture as a fungicide. The main advantage of using Thymus vulgaris as a pesticide is that it has very low toxicity and can rapidly decompose after it is applied on the plants thus safe for any other handlers or consumers who come in contact with the plant.

Thymus Vulgaris is a widely available Lamiaceae which is a native of meridional Europe, with its essential oil mainly composed of two phenolic monoterpenes, carvacrol and/or thymol, which are highly responsible for is antifungal activity. Fungi constitute the largest number of plant pathogens and are responsible for a range of serious plant diseases which damage plants by killing cells and/or causing plant stress. The organic or aqueous extract of Thymus Vulgaris can help in combating the fungal infections seen on various parts of the plants.

Some of the fungi are which are responsible for fungal diseases include downy mildews; powdery mildews; white blister, etc., There are several other fungi which causes diseases in the plants. The vegetable plants which are grown also get infected with some fungal diseases including anthracnose; botrytis rots; downy mildews; fusarium rots; powdery mildews; rhizoctonia rots; sclerotinia rots; sclerotium rots. The extract of Thymus Vulgaris with the properties can be used to combat the fungal disease. The extract of Thymus Vulgaris can actively act against the fungal infections which are spread on the plants.

For Example: If we consider Grape plant, the downy mildews; powdery mildews are the pathogens which affect the grape plant. In case of tomato plant, the most common fungal diseases that infect tomato plants include Anthracnose fruit rot; wilts, early blight, Septoria leaf spot, Fate blight, and Buckeye rot all which produce distinct symptoms. In case of the corn plant, the fungal infections such as such as grey leaf spot, northern com leaf blight, tar spot and common and southern rust, and so forth. With the help of the said synergistic composition, the diseases spreadon the grape plant can be treated.

The present invention discloses a synergistic composition which comprises of organic or aqueous extract of Annona Squamosa which acts as an insecticidal component. The Annona Squamosa extract or the custard apple (Common Name of Annona Squamosa) is a potential medicinal plant from the family Annonaceae, with diversified medicinal and pesticidal use, which can be cultivated in all tropical and subtropical countries including India. Annona Squamosa extract is used in the pesticide composition as an insecticidal agent which contains squamocin as an active ingredient.

According to the present invention, the Annona Squamosa seeds also contain acetogenins, which inhibits the development of the microbes on the plants and acts as insect control for plant sources. The extract of the Annona Squamosa is non-toxic to the animals but have pesticidal and / or insecticidal properties which can combat any insecticidal infections. The use of Annona Squamosa can be used on a wide variety of plants which can include pulses, millets, fibre crops, sugar crops, plantation crops, vegetables, fruits, spices, flowers, medicinal crops and so forth.

The present invention comprises of a component 5-Chloro-2-(4-chlorophenoxy) phenol which acts as an anti-bacterial component. 5-Chloro-2-(4-chlorophenoxy) phenol is a polychloro phenoxy phenol which is aromatic ether that is phenol which is substituted at C- 5 by a chloro group and at C-2 by a 2, 4-dichlorophenoxy group. It works as an antibacterial agent

According to the present invention, 5-Chloro-2-(4-chlorophenoxy) phenol acts as biocide specifically at lower concentrations (100 ppm- 500 ppm), which attacks multiple cytoplasmic and membrane of the bacteria, to eliminate the bacterial infection. 5-Chloro- 2-(4-chlorophenoxy) phenol at lower concentrations binds to bacterial enoyl-acyl carrier protein reductase enzyme which destabilizes it to combat the bacterial infections.

5-Chloro-2-(4-chlorophenoxy) phenol or 5-Chloro-2-(4-chlorophenoxy) phenol, which is the antibacterial ingredient which acts to slow or stop the growth of bacteria, fungi, and mildew. The use of 5-Chloro-2-(4-chlorophenoxy) phenol which can be directly be applied on the plants to prevent microbial growth.

The present invention comprises of another component Ethoxylated/Propoxylated or Ethoxylated-Propoxylated Natural Vegetable oil including but not limited to castor oil, Palm Oil, Coconut Oil, Soybean oil as natural surfactant. The said Ethoxylated/Propoxylated or Ethoxylated-Propoxylated Natural Vegetable oil molecules spontaneously bond with each other to form sealed bubbles as they have lower the surface tension. These surfactants may act as emulsifiers wherein the said surfactant is used in the synergistic composition as a part of formulation which creates an oil/water emulsion when the concentrate is added to water. These formulations are used when the active ingredient may not enter plant foliage readily due to high water surface tension or evaporation that leaves herbicides on foliage and results in no activity. The formulation components of the said composition includes Ethoxylated/Propoxylated or Ethoxylated- Propoxylated Natural Vegetable oil acts as a natural surfactant, can spontaneously bond with the all the components of the said composition. Additionally, any other surfactant which is having HLB (Hydrophile Lypophile Balance) of 9 to 20 can be used in the formulation described in the present invention.

The present invention comprises of another component which is natural oil and its derivatives from the sources like soyabean, cotton or palm. According to the present invention, the liquid formulations which are used in the said synergistic composition where the active ingredients are dissolved in the said natural oil and the surfactant is used in the said formulation so that it can be mixed with water for spraying. The natural oils and its derivatives like soya oil, cotton oil or palm oil which acts as stabilizers or solvents, which enables to preserve the desirable properties of the active ingredients of the said composition until it is prepared for desired application and applied on the plants.

According to the present invention, natural oil and its derivatives from the sources like soybean, cotton or palm, and 2-ethyl hexyl lactate/dimethyl sulfoxide/water/N, N Dimethyl decanamide and water can be used as a diluent is basically used to dilute the said synergistic composition. The said composition comprises of a diluent and an emulsifier which prevents the said composition from separating, and the diluent dilutes the pesticide to make it safer, more effective, and easier to measure, mix, apply, and handle.

According to the present invention, the said synergistic composition can be applied to the plants using to any type of common application equipment including but not limited to basic sprayer or a hydraulic sprayer, mist blowers and so forth. The said composition has high potential as an alternative to synthetic pesticides which are exiting in the market and their associated negative effects.

The synergistic composition as disclosed in the present invention can be prepared using various formulations which can include emulsifiable concentrate, micro- emulsion, oil dispersion, suspension concentrate, wettable powder, water dispersible granules and so forth.

Table 1: The formulation used to develop the emulsifiable concentrate of the synergistic composition

The process of formulating the emulsifiable concentrate of the synergistic composition is as follows: In laboratory mixer (DISPERMAT-R30 supplied by VMA-GETZMANN GMBH), the required quantity of Solvent/Diluent is weighed and added and mixed with continuous stirring; organic extract of Thymus Vulgaris is added in the mixer and stirred well until it dissolves, and while continuous stirring, the organic extract of Annona Squamosa is added and mixed until it forms clear solution. If required, the said mixture is heated to 40 to 45 degree Celsius. After the mixture has cooled down, 5-Chloro-2- 2(4-Chlorophenoxy Phenol) is added and stirred continuously until it completely dissolves; and the active matter of the composition is tested using HPLC (High performance liquid Chromatography/GC (Gas Chromatography).

Further, with continuous stirring, the ethoxylated derivatives of castor oil (Emulsogen EL-40 from Clariant, Agnique CSO 36 from BASF, Alkamuls 40 - Solvey) is added and mixed until it forms a clear solution. While stirring the solution continuously, the desired amount of Calcium alkyl benzene sulfonate (Agnique ABS-60-CE-H BASF, Ninate 70 B-Stepan Chemical Co, Rhodacal 60BR-Solvey) is weighed and added to the mixture, mixing it continuously by stirringuntil it is completely dissolved.

Table 2: The formulation is tested for physico-chemical properties as per below specifications

15 Accelerated Stability Studies @ 54 deg C (CIPAC MT 46.3)

Table 3:

The formulation used to develop the Micro emulsion formulation includes:

The process of the formulation of the micro emulsion formulation includes weighing desired amount of water (aqueous phase) in laboratory mixer (DISPERMAT-R30 supplied by VMA-GETZMANN GMBH) and adding Propylene Glycol (Kollisolv PG-BASF) (organic phase) and Defoamer (Agnique DFM-lll-S) and mix until homogeneous solution with constant stirring. This is followed by weighing desired amount of N, N Dimethyl decanamide (Agnique AMD-10-BASF) solvent in a separate beaker and stirring continuously to mix well, and in same beaker weigh desired amount of organic extract of Thymus vulgaris and mix till dissolves, and in same beaker weigh and add desired amount of organic extract of Annona Squamosa followed by 5-Chloro-2- (4-Chlorophenoxy) phenol and stir until they are completely dissolved. The product is then analysed for active matter and then proceeded for further addition of Soybean oil, followed by calcium alkyl benzene sulfonate(Rhodacal 65-BR-Solvey), and castor oil ethoxylate (Agnique CSO-36-BASF) and stir well until it completely dissolves. Then slowly add aqueous phase in to organic phase with stirring at high speed of 1500 to 2000 rpm to form white milky suspension of particle size less than 5 microns. To this suspension, add Xanthan gum 2% solution (Agropol-23 -Solvey) to modify viscosity.

The formulation tested for physico-chemical properties as per below specification.

The formulation for oil dispersion composition is:

The process of the formulation of oil-dispersion composition includes: In laboratory mixer (DISPERMAT-R30 supplied by VMA-GETZMANN GMBH) weigh desired amount of Methylated soybean oil and desired amount of Alcohol Ehtloylate (Lutensol TO 5-BASF) and calcium alkyl Benzene sulfonate (Rhodacal 6 BR- Solvey) and mix well to form dispersion along with constant stirring. Keeping the constant stirring, add Gerenol Odessy 5 supplied by solver and stir well to form homogeneous mix. Now at high speed of 1500 rpm slowly add organic extract of Thymus Vulgaris followed by organic extract of Annona Squmosa and -5- Chloro- 2-(4-Chlorophenoxy)Phenol and stir for 30 min. Once the suspension is ready, the formulation is completed by adding Rhemoer SC plus supplied by Solvey. The formulation is tested for physico-chemical properties as per belowspecification.

Accelerated Stability Studies @ 54 deg C (CIPAC MT 46.3) Table 9:

The formulation for suspension concentrates of the composition is:

The process of preparation of the said formulation includes: In a separate beaker equipped with homogenization (DISPERMAT-R30 supplied by VMA GETZMANN GMBH) facility weigh desired amount and DM water followed by Soprophor FLK, Agnique CP-72 L- Supplied by BASF and Pluronic PE 10500- Supplied by BASF with slow stirring. Weigh and add propylene glycol and Agnique DFM-111 -S-Supplied by BASF with constant stirring to form a solution. Increase the speed to 2000 rpm and slowly add kaolin clay to form a suspension. Weigh and add aqueous extract of Thymus Vulgaris followed by aqueous extract of Annona Squamosa and 5-Chloro-2-(4-Chlorophenoxy) Phenol and stir continuously to form greenish white suspension. This suspension is slowly feed to bead mill (DISPERMAT SL-B BEAD MILL supplied by VMA-GETZMANN GMBH USA for particle size reduction, ensuring the temperature of milling chamber and that to be product shouldn’t be raised to more than 25 deg C to have effective particle size reduction. Once the bead milling completed, the resulting product is transferred to a separate container, to which Agropol -23 is added by Solvey with constant stirring for 30 min, to complete the formulation.

The product specification of the said specification is: The product specification at Stability Studies at 54 Deg C discloses the following:

The wettable powder formulation of the said composition includes:

The process of preparation of the said composition includes: In a ribbon blender (Sanitary lab model 1 cu ft- Supplied by ROSS USA) which is charged with Tamol DN supplied BASF, Tamol FBPP Supplied by BASF and Supragil WP Supplied by Solvey were mixed together to form uniform mass. This mass is then fed to dry grinding mill air jet mill (Supplied by Promas machinery India), where the mixture is ground to the particle size of D-90- 20 microns and again fed to post blender and mixed for 1 hr to finish the dry blending part of the formulation. In a separate container, weigh and add Thymus Vulgaris organic extract, Annona Squamosa organic extract and 5-Chloro-2-(4-Chlorophenoxy) Phenol and mix well to form homogeneous solution/mixture. This mixture is then sprayed using the automated laboratory spray on the ground material which is holding in the previous ribbon blender to continue the blending operation for 2 hrs unless the mixture homogeneously to absorb the liquid active materials, and the product is analysed asper the specification.

The product specification of the said formulated product is:

Table 14

Storage stability @ 54 deg C for 14 days

The formulation for water dispersible granules is:

The process of formulation of the said product includes: The ribbon blender (Sanitary lab model 1 cu ft- Supplied by ROSS USA) is charged with Tamol DN supplied BASF, Tamol FBPP Supplied by BASF and Supragil WP Supplied by Solvey were mixed together to form uniform mass. This mass is then fed to dry grinding mill air jet mill (Supplied by Promas machinery India) where the mixture is ground to the particle size of D-90- 20 microns and again fed to post blender and mixed for 1 hr to finish the dry blending part of the formulation. In a separate container, weigh and add Thymus Vulgaris organic extract, Annona Squamosa organic extract and 5-Chloro-2-(4-Chlorophenoxy) Phenol and mixed well to form homogeneous solution/mixture. This mixture is then sprayed using the automated laboratory spray on the ground material which is holding in the previous ribbon blender continue this blending operation for 2 hrs unless the mixture homogeneously absorbs the liquid active materials, analyse the product as per the specification, and the mixture is then fed to Bextruder (Supplied Hosokawa-Japan) to form strands of extruded granules, water or any other organic lubricant can be added in to powder for better extrusion.

The product specification of the above formulation:

Storage stability @ 54 deg C for 14 days Table 18:

The present invention further discloses the bio-efficacy of the said synergistic composition showing the nematicide activity of various formulations.

Test 1: Nematicide Activity

The efficacy of nematicides viz with various formulations prepared was tested against some important genera of plant parasitic nematodes. All these formulations with plant origin active matters significantly suppressed populations of all the nematodes to varying degrees around roots of various plants including tomato, chilli, okra, etc., The emulsifiable concentrate (EC) Tl, Oil Dispersion (OD) T2 proved to be most effective in reducing the population of nematodes around Chilli, Tomato and Okra. Beneficial effects of these nematicides were evident by manifoldincrease in the yield or plant growth.

Materials and methods used in test: A field naturally infested with moderate populations of Meloidogyne incognita, Rotylenchulus reniformis, Tylenchorhynchus brassicae, Hoplolaimus indicus, Helicotylenchus indicus, Tylenchus filiformis and Trichodorus mirzai was prepared and divided into small beds, and later the field was irrigated and all the treatments were randomly arranged with three replications. After three weeks of treatment, the seeds of Okra (Abelmoschus esculentus Moench.) cv. Nirmal were directly sown and four-week- old seedlings, raised in autoclaved soil, of tomato (Lycopers'con lycopersicum (L) Karsten). cv.Bhagyashree, chilli (Capsicum annuum L.) cv. Pusha Sadabahar. Snowball were transplanted. Necessary cultivation practices were observed. Nematode populations of each treatment were determined with the help of Oostenbrink's elutriator and Baermann funnels (Southey, 1970), both prior to treatment as well as at the termination of the experiment (three months).

Results:

Table-20- Effect of Nematicide Formulation on population of Nematodes and Plant

Growth

Total parasitic % Reduction=Percent reduction of parasitic forms over untreated control .. Equation 1

Note:

Hop =Hoplolaimus indicus, Hel=Helicotylenchus indicus,

T yl=Tylenchus filifonnis, Mel=Meloidogyne incognita larvae, Tr i=Trichodorous mirzai Test 2: Minimum Inhibitory Concentration

Further, the formulations prepared as per the examples are tested for minimum inhibitory concentration (MIC) for gram-positive and gram-negative bacteria. In this test, the bio-efficacy of each formulation on plurality of gram-positive bacteria and plurality of gram-negative bacteria are tested, whose results are given below.

The below table discloses the efficacy of the formulation various gram-positive bacteria.

The below table discloses the efficacy of the formulation various gram-negative bacteria.

Note:

MIC- Minimum Inhibitory Concentration EC= EMULS IFIAB LE CONCENTRATE;

OD = OIL DISPERSION;

ME = MICRO EMULSION;

SC=SUSPENSION CONCENTRATE;

WP= WETTABLE POWDER;

WDG= WATER DISPERSIBLE GRANULES

Test 3: Fungicide Activity

Field experiment was conducted to study the efficacy of ametoctradin 300g/l + dimethmorph 225 g/1 SC against downy mildew disease of cucumber in farmer’s field. Cucumber variety used for the study was Ranebennur local. The soil type was red soil. The 30 days old cucumber plants were transplanted at the spacing of 90 cm x 75 cm. The plot size was 25-meter square. The cucumber crop was grown under irrigated condition. The experiment was conducted in randomized block design with 8 treatments and 3 replications. The treatments included were different formulations of fungicides along with untreated control. Observations on fruit as per plot were recorded. The data on disease severity and fruit yield were analyzed statistically. The treatment with Tl, T2, T3, T4, T5, T6 and TO serve as control. The dose 2 ml/liter and 3 ml/liter were used to evaluate the results, whereas TO control plot were treated with plain water. The observations were made after 30,45 and 60 days respectively. The results were showed that all the fungicide treatments were effective to reduce substantially the fungicide index, however the Tl (EC formulation), T2(OD formulation) and T3 (ME formulation) have showed better control over the other formulation and untreated control # 2 and 3 ml/liter of formulation, 3 ml / liter were substantially effective concentration to control check of fungicide index.

Bio efficacy of various formulations against Downy mildew disease of cucumber in rabi season during 2019-2020 @ 2 ml/liter Table 25:

Note:

TO: Control Tl: EC T2: OD T3: ME T4: SC T5: WP T6: WDG

Bio efficacy of various formulations against Downy mildew disease of cucumber in rabi season during 2019-2020 @ 3 ml/litre

The Percent Disease Index (PDI) was calculated using the formula below.

Percent Disease Index (PDI)= Sum of all disease ratings* 100 _

Total Number of ratings* Maximum disease grade

... Equation 2

Test 4: Insecticidal Activity

Insecticidal activity of Rhipiphorothrips cruentatus (grapevine thrips) using various formulation prepared on Adults and Nymphs

Reduction = (Total number of insects before treatment-number of living insects ^' ) * 100 percentage total number of insects before treatment

... Equation 3

Insecticidal activity of cabbage moth P. xylostella larvae using various formulation prepared on Adults and Nymphs

Insecticidal activity of gram borer Helicoverpa armigera using various formulation prepared

Table 29:

Examples of the said composition:

Example 1:

Organic/aq. Extract of Thymus vulgaris as anti-fungal component: 8.0% w/w Organic/aq. Extract of Annona Squamosa as insecticidal component: 10% w/w 5-Chloro-2-(4-chlorophenoxy) phenol as anti-bacterial component: 10% w/w Ethoxylated castor oil/Ethoxylated soybean oil another natural surfactant: 25% w/w Natural oil and its derivatives (Soya. Cotton/palm): 25% w/w 2, ethyl hexyl lactate/dimethyl sulfoxide/water/N, N Dimethyl decanamide: up to 100% w/w of total volume.