WO2012074486A1

WO2012074486A1 - Milk-based pesticidal formulation

Info

Publication number: WO2012074486A1
Application number: PCT/SG2011/000415
Authority: WO
Inventors: Melvin Donovan Pullen; Chek Chiew Chan; Su Min Charmaine Chan
Original assignee: Oro Agri International Ltd; Map Pacific Pte Ltd
Priority date: 2010-11-30
Filing date: 2011-11-25
Publication date: 2012-06-07

Abstract

A pesticidal composition, method of controlling pests and a method for improving the health and vigour of plants treated with the composition are disclosed. The composition comprises 20-90 parts by weight (pbw) milk (preferably fresh milk), 5 to 50 pbw surfactant, 0.5 to 10 pbw stabilizer to provide a satisfactory shelf-life, from 0 to 75 pbw water, and optionally 4 to 18 pbw of a high-terpene oil such as a citrus oil or a pine oil.

Description

MILK-BASED PESTICIDAL FORMULATION

This invention claims priority from South African provisional application ZA 2010/08580, filed on 30 November 2011.

FIELD OF THE INVENTION

This invention relates to crop protection and specifically environmentally friendly pesticidal formulations, and to their use in combating infestations of pests such as insects, bacterial and fungal growths. The invention is particularly, although not exclusively, concerned with pesticidal formulations and their use in agriculture, especially in the production of fruit and vegetables.

BACKGROUND TO THE INVENTION

In recent years, concerns have been raised about the potential danger of synthetic pesticides to humans and other non-target organisms. With this increased concern about toxicity and environmental safety, there has been renewed interest in the use of safer substances, including natural active ingredients, for pest control.

Much attention has been focused on the use of natural oils as a substitute for traditional insecticides with their attendant toxic and other dangerous side effects. These oils include horticultural oils that are highly refined petroleum products that can be mixed with water for application for control of target insect and mite pests without deleterious effects as a low toxicity alternative to broad spectrum insecticides. Since the mechanism of insect and mite control by means of spray oils is by suffocation and/or repellency of egg laying females, there is no requirement for the addition of toxic chemicals. These properties are a valuable and well recognized component of the practice of integrated pest management where oil spraying is intrinsically linked to natural control of pests by predators and parasitoids.

The application of oils also includes the application of high terpene oils, especially orange oil, together with a surfactant. Chelating agents have a long history of human use and are widely used in the food, cosmetic and pharmaceutical industries. European patent publication number EP1679002A1 describes a pesticide that comprises two main ingredients namely a first component in the form of a chelating agent, a metal complex of a chelating agent, or mixtures thereof, and a second component in the form of a carrier material. The latter is said to include dairy products that would naturally include milk and the main object of including the dairy product is as a bait to induce pests to ingest the pesticide the active ingredient of which is the chelating agent. The milk thus serves as an edible carrier material in the pesticidal composition.

Throughout the ages, milk has been used almost exclusively as a source of food for mammals, especially humans, and has received little attention in other fields of use. In more recent applications, prior art discloses the use of fresh milk, typically diluted with water as a fungicide, especially for the purpose of combating powdery mildew, botrytis and black spot. The efficacy in these applications are however rather low.

It will further be appreciated that due to the difference between insecticides and fungicides, it is not an obvious or expected that a specific ingredient would also be effective as an insecticide.

Although both fungicide and insecticide belong to the genus of pesticides, in practice fungicides are not used as insecticides. Fungicides and insecticides are very target specific and any one generally does not have an effect on the other type of organism. Both fungicides and insecticides target specific processes in fungi and insects respectively but as they differ in their physiology the targeted processes between fungi and insects differ.

Fungicides were developed to control the infection process and the growth of the hyphae amongst others. On the other hand most insecticides block the functioning of the nervous system or interfere with the moulting process of the insect. In fact, both fungicides and insecticides are often used on a specific fungus or insect and will not have a controlling effect on others in their respective groups. Insecticides for leaf miner and aphicides are one example of insecticides that are aimed at a specific insect. The same goes for fungicides for powdery and downy mildew that only target one of the two although they are both called mildews. This principle goes even further as for the same insect, different insecticides may target the larval stage and another the eggs.

It is well known that a healthy plant that grows with a minimum of physiological stress is more resistant to attack by pests. Insects will, for instance, target plants that they sense to be under stress due to environmental factors or internal stress due to nutritional status. Similarly, diseases are more inclined to infest and start developing in a plant when it comes under stress such as, but not limited to, during seed formation (pod-fill stage) or when the leaves reach maturity. Pests can also result in stress when the plant starts resisting attack by the formation of ethylene, which if present in sufficient quantities, can lead to premature senescence. A healthy plant can better overcome the negative effects of ethylene and prevent early onset of senescence. Often, the negative effect of a pest on a plant is more due to the early induction of senescence than the effect of the pest itself. Most vegetables and row crops are grown from seeds. Healthy, strong growth from the onset, starting at germination and through the seedling stage, leads to healthy strong plants later during their life cycle. A fast expanding foliage from the onset after germination of the seed leads to a better Net Assimilation Rate and a faster plant growth rate, which as the plant has a limited life span, is a very important determinant of the final production of the crop.

It has now surprisingly been found through inventive product engineering that the highly effective formulation(s) of the invention has both fungicidal and insecticidal properties and that it can be effectively applied in the germination of seeds and strengthening of the plant. It furthermore has a satisfactory shelf-life. The formulation can be prepared utilizing milk, or milk powder, as a starting ingredient and as at least one other ingredient.. It has also been found that a combination of milk and high terpene based oil compositions can result in an even more improved pesticidal formulation. The present invention provides a novel pesticidal composition, method of controlling pests and a method for improving the health and vigour of plants treated with the formulation of the invention. OBJECT OF THE INVENTION

It is an object of this invention to provide a pesticidal formulation including milk as at least one active ingredient. It is another object of the invention to provide a method of combating insect, mite, fungus, virus and bacterial attacks on plants using such a formulation.

It is another object of the invention to promote seed germination and seedling growth as well as further growth of the plants to better withstand attacks by pests and produce the maximum crop yield possible.

It is another object of the invention to provide for the combined use of a pesticidal formulation according to the invention and one or more other pesticidal formulations, especially a high terpene bio-pesticidal formulation.

SUMMARY OF THE INVENTION

In accordance with the invention there is provided a pesticidal formulation comprising fresh milk; a stabilizer/preservative water, and any other optional ingredients.

In accordance with the invention there is provided a pesticidal formulation comprising fresh milk; a stabilizer/preservative water, and a high terpene oil containing pesticide with any other optional ingredients. In accordance with one aspect of this invention there is provided a pesticidal formulation comprising from 20 to 90 parts by weight fresh milk; from 5 to 50 parts by weight surfactant; from 0.5 to 10 parts by weight stabilizer/preservative with a minimum quantity of stabilizer/preservative being selected to provide the formulation with a satisfactory shelf-life; from 0 to 75 parts by weight of water, and any other optional ingredients.

In certain embodiments, the compositions of the invention may include high terpene based oil compositions. Examples of high terpene (50% by weight or more) based oils, include but are not limited to citrus oils, pine oils, and other naturally occurring oils of plants that contain oil having 50% or more terpenes. Citrus oils include but are not limited to orange oil, lemon oil, lime oil, grapefruit oil and tangerine oil. Citrus oils may be obtained by any method from a citrus fruit. For example, citrus oil may be obtained from the skin or peel of a citrus fruit. Other methods of obtaining the citrus oil include but are not limited to cold pressing techniques.

Examples of surfactants include but are not limited to non-ionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants. Non-ionic surfactants include agents such as sorbitan monolaurate, sorbitan monopalmitate, sorbitan sesquioleate, sorbitan trioleate, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monostearate, polyethylene glycol monooleate, polyethylene glycol alkylate, polyoxyethylene alkyl ether, polyglycol diether, lauroyi diethanolamide, fatty acid iso-propanolamide, maltitol hydroxy fatty acid ether, alkylated polysaccharide, alkyl glucoside, sugar ester, oleophillic glycerol monostearate, self-emulsifiable glycerol monostearate, polyglycerol monostearate, polyglycerol alkylate, sorbitan monooleate, polyethylene glycol monostearate, polyoxyethylene sorbitan monooleate, polyoxyethylene cetyl ether, polyoxyethylene sterol, polyoxyethylene lanolin, polyoxyethylene bees wax, and polyoxyethylene hydrogenated castor oil; and the like. The alkyl glucosides are a class of non-ionic surfactants, which includes long chain alkyl glucosides or polyglucosides that are the condensation productsof (a) a long chain alcohol containing from about 6 to about 22, and preferably from about 8 to about 14 carbon atoms, with (b) glucose or a glucose-containing polymer. The alkyl glucosides have about 1 to about 6 glucose residues per molecule of alkyl glucoside. In one embodiment, a glucoside is decyl glucoside, which is the condensation product of decyl alcohol with a glucose polymer and is available commercially from Henkel Corporation of Hoboken, N.J. under the tradename, "Plantaren 2000." In another embodiment, the alkyl glucoside is AKZO-NOBEL AG 6210.

Anionic surfactants include agents such as sodium stearate, potassium palmitate, sodium cetyl sulfate, sodium lauryl phosphate, sodium polyoxyethylene lauryl sulfate, triethanolamine palmitate, polyoxyethylene sodium lauryl phosphate, and sodium N-acyl glutamate; and the like.

Cationic surfactants include agents such as stearyl dimethylbenzyl ammonium chloride, stearyl trimethyl ammonium chloride, benzalkonium chloride, and laurylamine oxide; and the like.

Amphoteric surfactants include alkylaminoethyl glycine chloride and lecithin; and the like.

Zwitterionic surfactants may optionally also be used. Zwitterionic surfactants are ammoniocarboxylates that are highly biodegradable.

The stabilizer/preservative for the milk, that is preferably cow's milk, would typically be a biocide selected from the 1 ,2-Benzisothiazolin-3-one or Benzyl Hemiformal family but could be any biocide having kinetics that would allow it to be used for the specific application.

The ingredients are preferably selected such that the pesticidal formulation conforms to the requirements of a biopesticidal formulation. Further features of this aspect of the invention provide for the pesticidal formulation to be in the form of a concentrate for use as is or for dilution preparatory to product application if a specific application so requires. In accordance with a second aspect of the invention there is provided a method of combating bacterial and insecticidal attacks on plants, the method comprising spraying on the foliage of the plants a formulation as defined above, diluted as may be appropriate with water. In accordance with a third aspect of the invention there is provided a method of combating pesticidal attacks on plants, the method comprising spraying on the foliage of the plants a fonnulation as defined above in admixture with a second biopesticidal formulation diluted as may be appropriate with water. Further features of the third aspect of the invention provide for the second biopesticidal formulation to be a high terpene-based biopesticidal formulation; and for the second biopesticidal formulation as typically described in US 6,277,389B1. For ease of reference these are defined as a High terpene containing natural oil as used herein means those natural oils having a terpene content of at least 50 per cent. It is preferable that the high terpene natural oil contains at least 90 per cent. Suitable high terpene containing natural oils includes oil from conifers such as citrus peel oils, preferably orange oil, grapefruit oil, lemon oil, or pine oil. Of these, orange oil is preferred and cold pressed orange oil the most preferred. Generally, the non-toxic pesticide composition contains from about 1 per cent to about 15 per cent by weight of high terpene containing natural oil, preferably from about 4 per cent to about 10 per cent by weight and more preferably from about 5 per cent to about 7 per cent by weight. Surfactants such as anionic and non-ionic surfactants are acceptable for use in the non-toxic aqueous pesticide of the present invention. Preferred are anionic surfactants such as salts of fatty acids, alkyl sulphates, alkyl ether sulphonates and alkyl aryl sulphonates. Examples of preferred surfactants include about 10 per cent sulfonic acid, about 6 per cent to about 7 per cent sodium laurel sulfate, from about 8 per cent to about 12 per cent alcohol ethoxylate and from about 1 per cent to about 2 per cent olefin sulfonate.

Generally, the non-toxic aqueous pesticide will contain from about 10 per cent to about 40 per cent by weight of surfactant(s), preferably from about 20 per cent to about 35 per cent by weight and more preferably from about 25 per cent to about 30 per cent by weight. The non-toxic aqueous pesticide may also contain various additives such as preservatives, pH neutralizes and/or clarifiers or stabilizers. Such preservatives may include butylated hydroxytoluene (BHT), p-Hydroxybenzoic acid, fungicide and bactericide.

The butylated hydroxytoluene (BHT) acts as an antioxidant. The antioxidant ranges from about 0.008 per cent to about 0.02 per cent by weight and more preferably about 0.01 per cent by weight.

The preferred non-toxic aqueous pesticide comprises about 6 per cent cold pressed orange oil, about 6.9 per cent sodium lauryl sulfate, about 11.6 per cent of alcohol ethoxylate, about 1.7 per cent sodium olefin sulfonate, about 10.2 per cent sulphonic acid, about 0.01 per cent butylate hydroxytoluene, about 0.58 per cent p-

Hydroxybenzoic acid, about 0.4 per cent bactericide, about 1.0 per cent fungicide, about 0.9 per cent urea and about 1.3 per cent sodium hydroxide caustic crystal with the balance a diluent such as water, all by weight. In accordance with a fourth aspect of the invention there is provided a method of treating seeds and improving germination and plant strength.

Soil insects can be extremely destructive to seed and seedlings, particularly when soil conditions are cold, wet, or do not favour rapid germination and emergence.

Seed treatments are used to protect seed from four major groups of pests: fungi and bacteria that cause seed; rots and decays, seedling blights, and smuts; soil insects, such as seed corn maggot and wireworms; storage insects, including weevils, moth larvae, and beetles; and birds and rodents that eat stored or planted seed and young seedlings. Treating seed with pesticides to protect it from pests that attack seed and seedlings can improve stand quality, increase yields, and increase return on investment. Germination is the process in which a plant emerges from a seed or spore, respectively, and begins growth.

In order that the invention may be more fully understood and expanded description thereof together with various test results will now follow.

DETAILED DESCRIPTION OF EXAMPLES OF THE INVENTION

In one embodiment of the invention, in a formulation herein termed ME, milk was employed as the sole active ingredient, and the formulation was prepared as a soluble liquid concentrate formulation being a water miscible formulation. The formulation is an environmentally friendly formulation in which all the liquid ingredients are blended into one final product.

In this particular embodiment of the invention the components were 80 parts by weight fresh cow's milk; 20 parts by weight surfactant such as selected from the organo silicone surfactant family (See Table i) with a suitable activity; and 1 part by weight biocide such as selected from the 1 ,2-Benzisothiazolin-3-one or Benzyl Hemiformal family as a stabliser/preservative. Whatever stabilizer is used, it should be suitably compatible with the other ingredients and should be effective to provide the formulation with a suitable shelf-life at room temperatures; a two-year shelf-life commonly being acceptable for pesticide registration requirements.

The formulation is prepared by ad-mixing the active ingredient, surfactant and biocide. The mixture is then stirred until homogenous.

It will be understood that the composition of cow's milk will vary from place to place and also according to the breed of cow and the season and it should be noted that tests may be necessary in order to confirm the suitability of any particular type of milk to the invention. Tests may also be necessary to determine accurately the proportions of the various ingredients that provide a satisfactory, and preferably an optimal, formulation.

It is well known that the makeup of milk is highly complex and milk contains numerous constituents including many vitamins. Whilst it is not yet known exactly what the active constituents of milk are, it is known that the content of lactoferrin and lactoperoxidase is, in applicants' view, likely to play an important role and the content of these constituents also depends on the origin of the milk. It is also within the scope of this patent that milk other than cow's milk could well be used. The patentee has tested the invention using goat milk, and powdered milk formulations and it is envisaged that most animal milk and milk formulations will be suitable as a constituent of the formulation.

It is also to be noted that, depending on the breed of animal from which the milk is obtained, the fat content can vary considerably with the fat content so for example milk from Jersey cows being as much as 40% higher than that from Holstein Frisian cows. In the prior art antifungal applications of raw milk it was suggested that the fat content can be reduced without in any way reducing the efficacy of the fungicidal activity of the milk. One of the main aims of such fat reduction was to avoid odours being created by fat that may go rancid, the milk being devoid of any preservative or stabliser.

In the instance of the present invention, the stabilizer/preservative should serve to avoid this but a decrease in fat content could well be beneficial from the point of view of reducing the quantity of fat applied to the foliage of a plant and also to usefully apply butterfat to its customary uses and market that which may otherwise be wasted.

In order to apply the pesticidal formulation described above, the formulation described is diluted (about 3 to 7 ml of ME with 1 litre of water) and sprayed directly on the target.

Typical insects that may be treated may include Thrips, Spodoptera Litura, Stem Borer, Green Leaf Hopper, Brown Plant Hopper, White Backed Plant Hopper, Mealy bug, Fruit Borer, Flea Beetle, Aphids, Whitefly, and Armyworm. Typical fungi that may be treated include Panicle Blast Disease, Anthracnose Disease, Powdery Mildew, Downy Mildew, Botrytis, and Sooty Mould. This list is not exhaustive and can be extended as more tests are done and more results are observed.

It has been found that the pesticidal formulation described above has multiple actions, these being insecticidal, bactericidal, fungicidal and plant growth enhancing.

Application of the pesticidal formulation has thus been observed to increase plant health and yield for agricultural products. The pesticidal formulation has also been found to be effective for seed treatment purposes to enhance germination of the crop, improve the yield and quality of the plant as well as reducing disease and insect incidence. This effect has been enhanced by adding ME with conventional products as well as ME and an orange oil containing pesticide; it also has shown to increase the efficacy and plant health properties of plants.

Milk contains a variety of carbohydrates, proteins, minerals and vitamins which will be absorbed by the plant. Through the vitamins, minerals and proteins absorbed into the plants, the plant's health is improved and hence there is a reduction in plant stress. The contribution towards the plant growth enhancing is believed to be primarily the proteins present in milk which are absorbed through the plants foliage in the manner of a foliar fertilizer thereby enhancing plant growth. It is believed that milk can improve the immunity of plants to disease and attack by bacteria and insects due to its constituents.

As a result of the fact that the product has more than one activity, farmers can reduce the number of pesticides sprayed onto their crops. This naturally reduces pesticide residue levels, another significant improvement over the existing procedure. Bioproducts such as the pesticidal formulation according to the invention, on the other hand, typically have no pesticidal residue levels compared to conventional products. In tests conducted to date, the performance of the embodiment of the invention described above on its own was tested in comparison with existing pesticides, namely, CONFIDOR™ a commercially available pesticide from Bayer with the active ingredient Imidacloprid; PROCLAIM™ 1.9EC a commercially available pesticide from Syngenta with the active ingredient Emamectin Benzoate; ANTHRACOL™ 70WP 2KH from Bayer with active ingredient Propineb; and RIDOM il™ gold 68 WP 2 KH from Syngenta with active ingredient Metalaxyl-M.

The results showed that the performance of the embodiment of the invention according to the invention was equivalent or better than that of the commercially available pesticide and, of course, without any of the environmental issues associated with such commercially available pesticides.

Combinations of the embodiment of the invention described above together with an orange oil containing pesticide such as that described in US patent no 6,277,389 and various other combinations were also tested. The results of these tests revealed that the combination of the two environmentally friendly pesticides performed better than either did on its own, especially when used in a ratio of about 5 (ME) to 3 (orange oil containing pesticide).

Indeed it was found that whereas the efficacy of the ME pesticidal formulation according to this invention when used on its own was up to about 80%; the efficacy of the orange oil containing pesticide was up to about 60%; whilst that of the combination in the ratio 5 (ME) to 3 (orange oil containing pesticide) showed an efficacy of up to about 90%. It was also noted that the plant health and strength were enhanced when the combination of the two pesticides was employed.

The invention therefore provides not only a highly useful environmentally friendly pesticide based on milk, but also a novel combination of pesticides both of which are environmentally friendly that has an even better efficacy.

Numerous variations are possible within the scope of the invention without departing from the scope hereof and the following non-limiting examples serve to further illustrate the invention: In all the examples the following definitions shall apply:

MS: Orange oil based product

ME: First embodiment of the invention: Milk product

MES15: Second embodiment of the invention (combination of MS and ME)

Table 1 : Organo-Silicone Surfactants

PRODUCT NAME MANUFACTURER/ ADJUVANT PRINCIPAL FUNCTIONING USE COMMENTS

DISTRIBUTOR CATEGORY AGENTS RANGE

ACTION 99 UCPA LLC Organo- Nonionic organosilicone

Silicone wetting agent

Surfactant

BREAK-THRU Plant Health Organo- Polyether- see 100% nonionic

Technologies Silicone polymethylsiloxane- label organosilicone,

Surfactant copolymer wetter, spreader, surfactant

CADENCE KALO, Inc. Organo- Polyether- 6-16 Approved for use

Silicone polymethylsiloxane- oz/100 with Regiment Surfactant copolymer and nonionic gal herbicide

and surfactant

Nonionic

Surfactant

CHEMPRO S-163 Chemorse, Ltd. Organo- Alkylphenol ethoxylate, 0.03- Silicone polyether-modified 1%

Surfactant polysiloxane, and glycol

blend

CHEMPRO S-172 Chemorse, Ltd. Methylated or Proprietary blend of 0.125- Ethylated polyalkyleneoxide modified 0.75%

Vegetable polydimethylsiloxane,

Oil nonionic emulsifiers and

and methylated seed oil

Organo- Silicone

Surfactant

DRENCH Garrco Products, Organo- Siloxane surfactant 100% 2-12 pH stable for up to

Inc. Silicone oz/100 4 hours, reduces

Surfactant gal surface tension

DYNE-AMIC Helena Chemical Methylated or Proprietary blend of 0.375- Blend of

Co. Ethylated polyethoxlated dimethyl 0.75% organosilicone

Vegetable siloxanes, alkylaryl surfactants and on ethoxylates and methylated methylated seed

and

Organo- Silicone

Surfactant

SYLGARD 309 Wilbur-Ellis Organo- Organosilicone 0.125- 100% active

Company Silicone 0.375%

Surfactant

THOROUGHBRED Estes, Inc. Organo- Propietary blend of 6-64 Nonionic silicone

Silicone polyalkyleneoxide modified oz/100 blend

Surfactant polydimethylsiloxane and gal

and nonionic surfactants

Nonionic

Surfactant

WIDESPREAD Loveland Products, Organo- Polyether- see Silicone surfactant MAX Inc. Silicone polymethylsiloxane- label

Surfactant copolymer, polyether

Example 1 : Efficacy Test and Farmer Field Practice treatments on Cabbage White Butterfly (Pieris rapae) of Cabbage Crop

Target crop : Cabbage

Days to maturity : 60 - 70 days

Target disease : Cabbage White Butterfly

Design : 6 Treatments x 4 Rep. (6 x 4 RCB)

Time of application : 25 days, 35 days and 45 days after sowing

Treatments

T1 = Control

T2 = MS 6% SL alone 80 ml/16 Lit

T3 = ME alone 80 ml/16 Lit

T4 = ME H30 alone 80 ml/16 Lit

T5 = ME (50 ml) + MS (30 ml) 80 ml/16 Lit

T6 = Fipronil (Conventional pesticide) 40 ml/16 Lit .Before application, at 25 days after sowing, the white butterfly infestation on the cabbage was severe. After 3 applications, there were no insects in the plots treated with MES15. The remaining treated plots showed a few insects, whereas in the Untreated Control plots insect control was substantially less.

MES15 resulted in the best control compared with the Untreated Control (100%), ME S15-H30 was second (85%), ME was 75%, Fipronil 80% and MS resulted in 70% control (See Table 1 below).

All the treated plots were slightly different on average weight of 10 cabbages in the treatments with the Control plots producing the smallest cabbages (Table 1).

All spray programs resulted in insect control with the combination MES15 (MS: ME products of 5:3) resulting in the 100% control compared with the individual products. The fact that this combination resulted in better control of the insects than the ME products alone, which were applied at a higher rate than in the combination, was surprising and not expected. The MES15 performed better than the conventional pesticide Fipronil.

Table 1. Efficacy Test of Map products and Farmer Field Practice on

Cabbage White Butterfly (Pieris rapae) and Yield

Mortality (%)

Rate Yield

Sr. No. Treatments After 3

(16 Lit) (Average weight of applications

10 cabbages)

3 5 10

(Lbs)

DAA DAA DAA

1 Control - - - - 2.88

2 MS 6% SL alone 80 ml 50 60 70 4.38

3 ME alone 80 ml 50 55 75 4.61

4 ME-H30 alone 80 ml 55 65 85 4.50

MES15 (ME: MS =

5 50 ml + 30 ml 80 90 100 4.75

5:3)

6 Fipronil 40 ml 60 75 80 4.57 Example 2: Efficacy Test of products and Farmer Field Practice Treatments on Late Blight (Phytophthora infestans) of Cherry Tomato

Days to maturity : 150 days

Design : 6 Treatments x 4 Rep. (6 x 4 RCB) Spraying time : Before Flowering time, flowering time and fruit setting time (3 applications)

Treatments T1 = Control

T2 = MS 6% SL alone 80 ml/16 Lit

T3 = ME alone 80 ml/16 Lit

T4 = ME H30 alone 80 ml/16 Lit

T5 = ME (50 ml) + MS (30 ml) 80 ml/16 Lit

T6 = Mancozeb (F.F.P) (Conventional pesticide) 60 gm/16 Lit

At harvest time, after 3 spray applications, all the treated plots showed good disease control compared with the Untreated Control (Table 2). MES15 (ME: MS = 5:3) 80 ml/16 treatment resulted in the lowest disease incidence of all the treatments. ME- H30 alone was (3.40%), ME alone (5.6%), MS 6% SL alone (9.5%) and conventional fungicide (10.3%) decreased disease incidence of late blight in this order. The finding that MES15 (ME: MS = 5:3) was superior in disease control although the ME content was only 62.5% of the individual treatment, was surprising. Even more surprising was that the treatment was decidedly more effective than the standard fungicide, mancozeb, which also has the disadvantage that it is an endocrine disruptor.

Results (Table 3) showed that among the treatments, MES15 (ME: MS = 5:3) had statistically significantly better effect on yield compared with all the other treatments. The yield results of ME-H30 and ME were lower but very similar to each other. MS treatment was next on yield. All treated plots had significantly more yield than the Untreated Control. Likewise, in examining the effect of the treatments on plant height, MES15 (ME: MS = 5:3) treatment resulted in the highest plants which differed statistically significantly from all the other treatments. The rest of the treatments followed the same pattern of statistical differences as yield (Table32). No phytotoxic effect on the crop was found.

All treatments resulted in disease control with MES15 (ME: MS = 5:3) by far superior in this respect. The plants treated with MES15 (ME: MS = 5:3) also had the highest yield and plant height.

Table 2. Efficacy Test of 5 Types of products and Farmer Field Practice on

Late Blight (Phytophthora infestans) of Cherry Tomato

Sr. No. Treatments Rate Disease Incidence (%)

BF 5 DAA 25 DAA 45 DAA

1 Control - 20.8 35.62 40.5 50.1

80 ml/16

2 MS 6% SL alone 20.9 15.45 12.3 9.5

Lit

80 ml/16

3 ME alone 25.8 18.3 10.6 5.6

Lit

80 ml/16

4 ME - H30 alone 20.9 14.5 10.1 3.4

Lit

80 ml/16

5 MES15 (ME: MS = 5:3) 20.3 10.8 5.8 1.2

Lit

60 gm/16

6 Mancozeb (F.F.P) 25.6 20.2 15.8 10.3

Lit Table 3. Efficacy Test of products and

Farmer Field Practice Treatments on Cherry Tomato Yield and Plant Height

Coefficient of Variation S_x = Standard Deviation

□□ = Highly Significant

Example 3: Efficacy Test and Farmer Field Practice Treatments on Bacterial Wilt {Pseudomonas solanacearum) of Sweet Chilli

Days to maturity : 110 - 120 days

Target disease : Bacterial Wilt

Design : 6 Treatments x 20 plants (Simple Trial)

Spraying time : Fruit setting time and fruiting time

Time of application : Two sprays starting at fruit setting and 10 days later. Treatments

T1 = Control

12 = MS 6% SL alone

T3 = ME alone

T4 = ME H30 alone

T5 = ME (50 ml) + MS (30 ml)

T6 = Metalaxyl 25% WP (Conventional pesticide)

The bacterial wilt infection on the plants was already serious (around 25%) at the time of the first spray. At harvest time, all the spray treatments had controlled the disease compared with the Untreated Control that showed 60% infection (Table 4). MES15 (ME: MS = 5:3) was the best for controlling the disease. Disease Incidence for ME and ME -H30 treatment was slightly less than that of MES15 treatment but all were still better than the conventional product.

MES15 (ME: MS = 5:3) treatment resulted in the best yield (5.92 lb/plant) and was statistically significantly better than the next 4 treatments. ME, ME-H30, MS and F.F.P, which did not differ significantly from on each other. The Untreated Control had significantly less yield than all the spray treatments. All treatments that had ME in the mixture resulted in significantly better plant height than MS or metalaxyl. The Untreated Control had significantly less height than all the other treatments (Table 4). No phytotoxic effect on the crop by spraying the above treatment rates MES15 treatment resulted in the best control of the disease and the highest yield was also obtained with this treatment. The result that the combination of the dairy product and the orange oil product resulted in the best control even though less of each had been used than in the individual product treatments was surprising and not expected. Table 4. Efficacy Test and Farmer Field Practice Treatments on Bacterial Wilt (Pseudomonas solanacearum) and Yield of Sweet Chili

Example 4: Russian aphid on wheat

Objective and methods:

A trial was conducted to test the effect of a MES15 on the control of Russian wheat aphid on winter wheat at Malmesbury, South Africa. Trail layout was a randomized complete block design with 4 replicates. Two spray applications on 15 and 22 August 2011 respectively with various combinations of the products individually and combined in different mixtures at a rate of 500 liter/ha were tested. This was compared with an industry standard pesticide application of Mospilan. Assessments were made at pre-treatment and 3, 7 and finally 14 days after the last application. An ANOVA was performed on the severity of infestation data and the percentage control by each treatment compared with the pre-treatment assessment was also determined.

Results

The results presented in Table 5 include the severity levels of the pest at pre- treatment and finally at 14 days after last application. The control obtained by each treatment compared with the pre-treatment severity is also provided.

Table 5. Severity of Russian wheat aphid infestation by various treatments after two sprays applications, one week apart.

a-d: Means accompanied by a common letter do not differ significantly differ at P=0.05

Discussion and conclusions:

All spray treatments resulted in statistically significant control compared with the Untreated Control which actually had an increase in severity of the pest over time. The product of the invention and ME alone at 800 ml all resulted in as efficient control of the pest compared with the Mospilan standard.

However, the product of the invention required a much lower amounts for efficient control than ME or MS on its own. Compare the 70% control obtained by ME at 800 ml with the 70% control obtained by only 105 ml: 195 ml) blend of ME and MS. This result is surprising, showing the higher efficiency of the product of the invention. Example 5: Seed treatment

Diseases of vegetable crops often are caused by fungi or bacteria that are carried on or with the seed.

Disinfection of the seed to kill these parasites is an important method of fighting vegetable diseases. In addition, the chemicals that remain on or in the seed help protect the seed and the developing seedling from decay or dam ping-off caused by fungi in the soil.

The effect of the product of the invention on the germination of seeds and growth of various types of vegetable plants were tested on cabbage, cauliflower, spinach and tomato.

Germination testing

A germination test determines the percentage of seeds that are alive in any seed lot. The level of germination in association with seed vigour provides a very good estimate of the potential field performance. While the speed of germination varies slightly across varieties, seeds should absorb moisture within 2 days and produce a root and the first leaf within 4 days. At this point the seed is considered to have germinated.

Germination testing is important as it is often the only test a farmer can conduct on seed to determine if it is suitable for planting. When seed is stored in traditional open systems the germination rate of most rice seed begins to deteriorate rapidly after 6 months. Also many varieties have a dormancy period immediately after harvest that can last for 1-2 months. By knowing the germination rate farmers can adjust their planting rates to attain the desired plant population in the field

Procedure

1. Cabbage, Cauliflower - Seed treatment

- Dilute 5 liter of MS + ME with 5 liter of water to spray on 6 kg of cabbage seeds or cauliflower seeds, then cabbage seeds are sowed in trays contain substrate

- 6 kg of seeds for 1 ha (6 kg equivalent to 30 000 seedlings)

- 5 liter of product for seed treatment on 1 ha

- It is customary for farmers to buy seedlings from local seed nurseries to plant and actual sowing of seeds rarely occurs.

2. Spinach - Seed treatment

- Dilute 2 liter of MS + ME with 20 liter of water, we sow 20 kg of seeds for 1 ha

- Method of seed soaking: dilutes product with 50 - 60°C warm water in bucket or basin then soaks seeds in 24 hours, after that picks seeds to drain and sow seeds in field.

- Because seed skin of spinach is very hard so we must soak seeds to help seeds can germinate better

- 2 kg of seeds for 1000 m², 20 kg of seeds for 1 ha

- 200 ml of product for 1000 m², 2 liter of product for 1 ha

3. Tomato - Seed treatment.

- Dilute 50 ml of MS + ME with 1 liter of water to spray on 30 g of tomato seeds, then allow seeds to drain, after that sows seeds in trays containing the substrate

- 30 g of seeds for 1 ha

- 50 ml of product for seed treatment on 1 ha.

- It is customary for farmers to buy seedlings from local seed nurseries to plant and actual sowing of seeds rarely occurs. Calculating the germination rate

Germination rate is the average number of seeds that germinate over the five and ten day periods.

Germination (%) = Number of seeds germinated x 100

Number of seeds on tray

Example 5.1 : Cabbage - Seed treatment (Figure 1)

The product of the invention showed a significantly higher germination rate than the conventional product. At 21 DAS, plants in treatment were a little bit higher and greener than the seeds treated with the conventional product.

Example 5.2: Spinach - Seed treatment (Figure 2) Table 6: Rating of plant characteristics: 1 best, 10 worst

Both product of the invention and the conventional treatments showed a small improvement in plant height compared with the untreated plants. The product of the invention however resulted in the stems being thicker and the leaves greener than in the untreated plants and/or conventional product treated plants.

Figure 3: Seedling at 26 days after sowing Example 5.3: Cauliflower - Seed treatment (Figure 4)

Both product of the invention and the conventional treatments showed a small improvement in plant height compared with the untreated plants. Both product of the invention and the conventional treatments, plants were higher and the stems thicker than plants in untreated plots.

Example 5.4: Tomato - Seed treatment (Figure 5)

Table 7: Rating of plant characteristics: 1 best, 10 worst.

Figure 6: Seedlings at 10 days after sowing

Both product of the invention and the conventional treatments, plants were higher, stems were thicker and the leaves were greener than that of untreated plants. The conventional product showed the best in germination rate, followed by the product of the invention.

Example 5.5: Comparative treatment: cabbage - tomato - spinach (Figure 7)

CONCLUSIONS ON ALL TRIALS:

In repeated trials, the product of the invention showed the best in germination rate, promoting plant development. Plots, plants leaves are thicker and greener in the product of invention treatments than plants treated with the conventional products and untreated plots.

Claims

1. A pesticidal formulation comprising from 20 to 90 parts by weight milk, from 5 to 50 parts by weight surfactant, from 0.5 to 10 parts by weight stabilizer with a minimum quantity of stabilizer/ being selected to provide the formulation with a satisfactory shelf-life; from 0 to 75 parts by weight of water, and any other optional ingredients.

2. A pesticidal formulation as claimed in claim 1 wherein the milk is fresh milk.

3. A pesticidal formulation as claimed in either one of claims 1 or 2 which includes as an ingredient, high terpene oil.

4. A pesticidal formulation as claimed in any one of claims 1 to 3 wherein the formulation includes from 4 to 18 parts by weight of a terpene containing pesticidal composition.

5. A pesticidal formulation as claimed in either one of claims 3 or 4 wherein a high terpene based oil includes one or more of citrus oils, pine oils, and other naturally occurring oils of plants that contain oil having 50% or more terpenes.

6. A pesticidal formulation as claimed in claim 5 wherein the terpene containing pesticidal composition contains between 2 and 15% citrus oil.

7. The formulation of any of the preceding claims wherein the surfactants include a surfactant selected from anyone of a non-ionic surfactants, anionic surfactants, cationic surfactants, zwitterionic and amphoteric surfactants

8. A pesticidal formulation as claimed in any one of the preceding claims wherein the surfactant includes at least one non-ionic surfactant selected from sorbitan monolaurate, sorbitan monopalmitate, sorbitan sesquioleate, sorbitan trioleate, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monostearate, polyethylene glycol monooleate, polyethylene glycol alkylate, polyoxyethylene alkyl ether, polyglycol diether, lauroyl diethanolamide, fatty acid iso-propanolamide, maltitol hydroxy fatty acid ether, alkylated polysaccharide, alkyl glucoside, sugar ester, oleophillic glycerol monostearate, self-emulsifiable glycerol monostearate, polyglycerol monostearate, polyglycerol alkylate, sorbitan monooleate, polyethylene glycol monostearate, polyoxyethylene sorbitan monooleate, polyoxyethylene cetyl ether, polyoxyethylene sterol, polyoxyethylene lanolin, polyoxyethylene bees wax, and polyoxyethylene hydrogenated castor oil or organosilicon .

9. A pesticidal formulation as claimed in any one of the preceding claims wherein the surfactant includes at least one anionic surfactant selected from sodium stearate, potassium palmitate, sodium cetyl sulfate, sodium lauryl phosphate, sodium polyoxyethylene lauryl sulfate, triethanolamine palmitate, polyoxyethylene sodium lauryl phosphate, and sodium N-acyl glutamate.

10. A pesticidal formulation as claimed in any one of the preceding claims wherein the surfactant includes at least one cationic surfactant selected from stearyl dimethylbenzyl ammonium chloride, stearyl trimethyl ammonium chloride, benzalkonium chloride, and laurylamine oxide.

11. A pesticidal formulation as claimed in any one of the preceding claims wherein the surfactant includes at least one amphoteric surfactant selected from alkylaminoethyl glycine chloride and lecithin.

12. A pesticidal formulation as claimed in any one of the preceding claims wherein the stabilizer is a preservative.

13. A pesticidal formulation as claimed in any one of the preceding claims wherein the stabilizer is a biocide.

14. A pesticidal formulation as claimed in Claim 13 wherein the biocide is selected from the 1 ,2-Benzisothiazolin-3-one and Benzyl Hemiformal family.

15. A pesticidal formulation as claimed in any one of the preceding claims wherein the ingredients are selected such that the pesticidal formulation conforms to the requirements of a biopesticidal formulation.

16. A method of combating bacterial and insecticidal attacks on plants, the method comprising spraying on the foliage of the plants a pesticidal formulation as defined in any one of the preceding claims.

17. A method of combating pest attacks on plants, the method comprising spraying on the foliage of the plants a formulation as defined in any one of the preceding claims in admixture with a second biopesticidal formulation.

18. A method as claimed in claim 17 wherein the second biopesticidal formulation is a high terpene-based biopesticidal formulation.

19. A method as claimed in claim 17 wherein the second biopesticidal formulation comprises at least one surfactant selected from the group consisting of sodium laurel sulphate, alcohol ethoxylate, olefin sulphonate, sulphonic acid and mixtures thereof and at least one high terpene containing natural oil selected from the group consisting of citrus peel oils, pine oils and mixtures thereof, the pesticide containing from about 20% to about 35% of said at least one surfactant by weight per cent and from about 4% to about 10% of said at least one high terpene containing natural oil by weight per cent and from about 60% to about 70% of water by weight per cent.

20. A method of treating seeds and improving germination and plant strength, the method comprising treatment of the seeds with a pesticidal formulation as claimed in any one of the claims 1 to 15.