Pesticide product containing vegetable oil and essential oil as active ingredients, process for its preparation and its use

Abstract

The invention relates to a pesticidal composition comprising: a) as vegetable oil active ingredient at least three compounds selected from the group consisting of soybean oil, eugenol, cinnamaldehyde, tea tree oil, rapeseed oil, clove oil, spearmint oil; b) optionally, as inorganic active ingredient, Cu(I) oxide; c) as excipient, a compound selected from the group consisting of xanthan gum, polyvinyl alcohol, magnesium aluminium silicate, ethoxylated vegetable oil, preferably ethoxylated castor oil; benzenesulphonic acid C10-13 alkyl derivative, ethoxylated sorbitan ester, C12-14 ethoxylated alcohol, potassium polyarylphenyl ether phosphate, the sodium salt of methylene polynaphthyl sulphonate; (d) optionally glycerol as antifreeze; (e) optionally an antioxidant, preferably ascorbic acid; and (f) optionally water as a carrier, as required. The invention also relates to a process for the preparation of the composition according to the invention and its use as a pesticide.

Description

[0001] 729152 / MI

[0002] PESTICIDE PRODUCT CONTAINING VEGETABLE OIL AND ESSENTIAL OIL AS ACTIVE INGREDIENTS, PROCESS FOR ITS PREPARATION AND ITS USE

[0003] The invention relates to a pesticidal composition comprising: a) a vegetable oil as active ingredient, selected from the group consisting of soybean oil, eugenol, cinnamaldehyde, tea tree oil, rapeseed oil, clove oil, spearmint oil; b) optionally, as inorganic active ingredient, Cu(I) oxide; c) as excipient, a compound selected from the group consisting of xanthan gum, polyvinyl alcohol, magnesium aluminium silicate, ethoxylated vegetable oil, preferably ethoxylated castor oil; benzenesulphonic acid Cl 0-13 alkyl derivative, ethoxylated sorbitan ester, C12-14 ethoxylated alcohol, potassium polyarylphenyl ether phosphate, the sodium salt of methylene polynaphthyl sulphonate; (d) optionally glycerol as antifreeze; (e) optionally an antioxidant, preferably ascorbic acid; and (f) optionally water as a carrier, as required. The invention also relates to a process for the preparation of the composition according to the invention and its use as a pesticide product.

[0004] BACKGROUND ART

[0005] In our prior art search, we found that the closest known solution is represented by international patent publication No. W02020198853 Al, which discloses an aqueous nanoemulsion composition comprising 0.05-55 wt% volatile oil, 0.04-65 wt% solvent, 0.01- 25 wt% sorbate, 0.00025-0.37 wt% saponin and water in the amount necessary to achieve 100 wt%. The essential oil in the composition is one or more selected from the following: aniseed oil, lemon oil, orange oil, oregano oil, rosemary' oil, wintergreen oil, thyme oil, lavender oil, clove oil, hops oil, tea tree oil, citronella oil, wheat oil, barley oil, lemongrass oil, cedar leaf oil, cedar wood oil, cinnamon oil, fleagrass oil, geranium oil, sandalwood oil, violet oil, cranberry oil, eucalyptus oil, vervain oil, peppermint oil, basil oil, fennel oil, fir oil, balsam oil, oregano oil, goldenseal (Hydrastis canadensis) oil, Berberidaceae daceae oil, Ralanhiae oil, turmeric (Curcuma longa) oil, sesame oil, macadamia nut oil, evening primrose oil, coriander oil, pimento berries oil, rose oil, bergamot oil, rosewood oil, chamomile oil, sage oil, clary sage oil, cypress oil, sea fennel oil, frankincense oil, ginger oil, grapefruit oil, jasmine oil, juniper oil, lime oil, mandarin oil, mar oram oil, myrrh oil, neroli oil, patchouli oil, pepper oil, black pepper oil, petitgrain oil, pine oil, rose oil, spearmint oil, spikenard oil, vetiver oil, a conifer essential oil, ylang ylang or combinations thereof. However, the embodiments do not include combinations of the examined oils. This preparation optionally contains ascorbic acid. The composition is recommended for the control of fire blight, powdery / mildew, septoria and botrytis on crops including, among others, apple, pear, potato and tomato.

[0006] The fungicidal activity of the essential oils used as active ingredients in the tested composition is already known, for example, US5679351A discloses a fungicidal plant protection product containing clove oil.

[0007] The inhibitory' effect of spearmint oil against Macrophomina phaseolina, which causes root rot of cotton plants, is described by El-Shoraky et al. in Antifungal Activity of Spearmint and Peppermint Essential Oils against Macrophomina Root Rot of Cotton, J. Plant Prot. and Path., MansouraUniv., Vol.9 (11): 775-781, 2018; and the activity of tea tree oil against Botrytis cinerea is described by Wang et al. in Antifungal modes of action of tea tree oil and its two characteristic components against Botrytis cinerea, J. Applied Microbioligy, Volume! 19, Issue 5, November, pp 1253-1262.

[0008] The fungi cidal / bacteri ci dal activity of the combination of the above essential oils is described in EP1534076A1 for tea tree oil and clove oil, in CN104224923A for clove oil and spearmint oil, and in CN104224923A for tea tree oil and spearmint oil.

[0009] Other, more general antibacterial applications are described in US2011038964A1 for a preparation for the microb act eri l removal of microbes from solid surfaces containing as active ingredients one or more oils selected from the group consisting of rosemary' oil, tea tree oil, spearmint oil, peppermint oil, clove oil, lemongrass oil, cedar oil and cinnamon oil.

[0010] WO2011117891A2 discloses an environmentally friendly fungicidal composition for the protection of rubber plantations, which comprises vegetable oils or their esters, for example soybean or rapeseed oil, a mineral oil derivative and fungicidal copper salt.

[0011] US 10905114B2 relates to an arthropodicidal preparation containing benzyl alcohol and essential oils (e.g. cinnamon and clove oil) and various detergents such as ethoxylated castor oil, ethoxylated alcohol, and lauryl sulphate etc.

[0012] US2001055628A1 discloses a synergistic pesticide concentrate containing volatile and non-volatile oils, wherein the non-volatile oil may be soybean oil or rapeseed oil or other vegetable oil, and the volatile oil may be for example cinnamon oil or clove oil; and as an emulsifier it comprises lauryl sulphate.

[0013] WO2023132811 Al relates to a composition comprising microencapsul ted essential oils and chitosan, wherein e.g. cinnamic aldehyde and / or eugenol are included as active ingredients in addition to detergents, carrier oil and emulsion stabilizer and other excipients. The preparation is for use against the following plant pests: Aspergillus niger, Penicillium digitatum, Botrytis cinerea, Xanthomonas juglandis, Xanthom.onas phaseoli, Clavibacter michiganensis, Fusarium culmorum, Rhizoctonia solani, Sclerotinia sclerotiorium, Ascohyta rabiei.

[0014] In addition to those listed above, there are numerous publications on the use of plant oils or essential oils, alone or in combination, for plant protection:

[0015] Wurms et al. in Fungicidal Activity of Soybean Oil against Powdery Mildew on Wheat (http: / / dx.doi.org / 10.5772 / intechopen.81516), Baker et al. in Soybean Oil Profile: Active Ingredient Eligible for Minimum Risk Pesticide Use (http: / / hdl.handle.net / 1813 / 56142), and the Peer review of the pesticide risk assessment of the active substance rape seed oil (EF SA Journal 2022;20(5):7305; 10, 2903 / j.efsa.2022.7305) analyse the issues of the applicability of rape seed oil.

[0016] Thabet et al. in Antifungal Activities of Clove Oil Against Root Rot and Wilt Pathogens of Tomato Plants, American -Eurasian J. Agric. & Environ. Sci., 18 (3): 105-114 2018; 10.5829 / idosi.aejaes.2018.105.114; Huang et al. in Effect of clove oil on plant pathogenic bacteria and bacterial wilt of tomato and geranium, journal of Plant Pathology (2010), 92 (3), 701-707; Kumar et al. in Greenhouse and field experiments revealed that clove oil can effectively reduce bacterial blight and Increase yield in pomegranate. Food Energy Secur. 2021;10:e305.; https: / / doi.org / 10. I002 / fes3.305; and Mahmoud et al. in Laboratory and Semi - Field Evaluation and Effect of Clove Essential OU against Two - Spotted Spider - Mite Tetranychus urticae, Koch. (Acari:Tetranychidae); J. of Plant Protection and Pathology, Mansoura Univ., Vol. 13 (2):59 - 61, 2022 discuss the effects of eugenol (or clove oil), the latter two of these documents report greenhouse and outdoor applications.

[0017] Abbo et al. in The Response of Tea Tree Oil as a Biofungicide Against Early Blight Disease in Tomato Crop (Solanum Lycopersicum in Sudan T Conference on International Research on Food Security, Natural Resource Management and Rural Development, Tropentag 2009 University of Hamburg, October 6-8, 2009; Hendges et al. in Antifungal activity and control of the early blight in tomato through tea tree essential oil. Crop Protection 148 (2021) 105728 https: / / doi.Org / 10.1016 / j.cropro.2021.105728; Dalio et al. in Tea Tree Oil Induces Systemic Resistance against Fusarium wilt in Banana and Xanthomonas Infection in Tomato Plants 2020, 9, 1137; doi:10.3390 / plants9091137; and Yasin et al. in River Tea Tree Oil: Composition, Antimicrobial and Antioxidant Activities, and Potential Applications in Agriculture Plants 2021, 10, 2105. https: / / doi.org / I0.3390 / plants l0102105 disclose a study of the effects of tea tree oil, while Shen et al. in Application of cinnamaldehyde for the postharvest storage of fresh horticultural products, Horticult. Int. J. 2021;5(3):103-105. and Kowalska et al. in Cinnamon as a Useful Preventive Substance for the Care of Human and Plant Health, Molecules 2021 :26:5299. htps: / / doi.org / 10.3390 / molecules26175299 disclose a study of the effects of cinnamic acid aldehyde (cinnamaldehyde or cinnamonoil).

[0018] For further general overviews on the role of essential oils in plant protection, see Chang et al. in Biocontrol Potential of Essential Oils in Organic Horticulture Systems: From Farm to Fork Frontiers in Nutrition | www.frontiersin.org; January 2022 Volume 8 Article 805138 and Breiing et al. in Fungicidal Efficacy of Drying Plant Oils in Green Beans against Bean Rust Uromyces appendiculatus) Plants 2021, 10, 143. https: / / doi.org / 10.3390 / plantsl0010143.

[0019] A synergistic composition is described by Soe et al. in Synergistic effect of sesame oil and clove oil on toxicity against the pulse beetle, Callosobruchus maculatus (Fabricius) fColeoptera: ChysomelidaeZ KHON KAEN AGR. J. 47 SUPPL.1 (2019): this describes a combination of sesame oil and clove (essentially eugenol), while US2001055628A1 discloses a pesticidal concentrate containing volatile and non-volatile oils with synergistic activity, where the non-volatile oil can be soybean oil, rapeseed oil, or an other vegetable oil, and the volatile oil can be e.g. cinnamon oil (essentially cinnamon aldehyde) or clove oil (essentially eugenol); and also comprises lauryl sulphate as an emulsifier.

[0020] A combination formulation without the mention of synergistic effect is disclosed in CN103783093B, which also includes clove oil or cinnamon oil as active ingredient, in addition to rapeseed oil or esterified soybean oil.

[0021] It can be stated that no document relating to a composition identical or equivalent to the composition of the present invention was found during the search. Although the beneficial effects of individual components, or combinations of components, have been described in state- of-the-art documents, these are only basic observations and do not necessarily provide an incentive for those skilled in the art to pursue further research into developing a new, more effective composition containing more than two vegetable oil components.

[0022] BRIEF DESCRIPTION OF THE FIGURES

[0023] Figure 1 Inhibitory effect of the essential oil mixture on the growth of Fusarium graminearum, Fusarium culmorum and Phytophthora infestans strains in Petri dishes after 7 days.

[0024] Figure 2 Inhibitory effect of the essential oil mixture on the growth of different strains in Petri dishes after 7 days.

[0025] Figure 3 Growth of Fusarium graminearum strain in media with different concentrations of the active ingredient (0, 0.125, 0.250, 0.500, 0.750, 1.125 V / V%).

[0026] Figure 4 Growth of Fusarium culmorum strain in media with different concentrations of the active ingredient (0, 0.125, 0.250, 0.500, 0.750, 1.125 V / V%).

[0027] Figure 5 Growth of Phytophthora infestans strain in media with different concentrations of the active ingredient (0, 0.125, 0.250, 0.500, 0.750, 1.125 V / V%).

[0028] Figure 6 Growth of Monilia laxa strain in media with different concentrations of the active ingredient (0, 0.125, 0.250, 0.500, 0.750, 1.125 V / V%).

[0029] Figure 7 Growth of Monilia fructicola strain in media with different concentrations of the active ingredient (0, 0.125, 0.250, 0.500, 0.750, 1.125 V / V%).

[0030] Figure 8 Growth oiMonilia fructigena strain in media with different concentrations of the active ingredient (0, 0.125, 0.250, 0,500, 0.750, 1.125 V / V%).

[0031] Figure 9: Growth of Botrytis cinerea strain in media with different concentrations of the active ingredient (0, 0.125, 0.250, 0.500, 0.750, 1.125 V / v%).

[0032] Figure 10 Growth of Fusarium oxysporum strain different media with different concentrations of the active ingredient (0, 0.125, 0.250, 0.500, 0.750, 1.125 V / v%).

[0033] Figure 11 Growth of Fusarium solani strain in media with different concentrations of the active ingredient (0, 0.125, 0.250, 0.500, 0.750, 1.125 V / v%). Figure 12 Growth of Fusarium sambucinum strain in media with different concentrations of the active ingredient (0, 0.125, 0.250, 0.500, 0.750, 1.125 V / v%).

[0034] Figure 13 Growth of Sclerotinia sclerotiorum strain in media with different concentrations of the active ingredient (0, 0.125, 0.250, 0.500, 0.750, 1.125 V / v%).

[0035] Figure 14 Microscopic images of Botrytis (A) and Monilia (B).

[0036] Figure 15 In vitro efficacy of essential oils and their mixtures against Monilia laxa

[0037] Figure 16 In vitro efficacy of essential oils and their mixtures against Monilinia fructigena

[0038] Figure 17 In vitro efficacy of essential oils and their mixtures against the pathogen Fusarium oxysporum.

[0039] Figure 18 In vitro efficacy of essential oils and their mixtures against the pathogen Fusarium solani.

[0040] Figure 19 Photo documentation of the efficacy trial series of the compositions with three active ingredients.

[0041] THE TECHNICAL PROBLEM TO BE SOLVED BY THE INVENTION

[0042] The technical problem to be solved by the present invention is to provide a pesticide that comprises vegetable and essential oils and exhibits exceptional activity against plant pathogens.

[0043] INSIGHT UNDERLYING THE INVENTION

[0044] It is recognized that outstanding pesticide efficacy can be achieved by the application of the active ingredients according to the invention.

[0045] BRIEF DESCRIPTION OF THE INVENTION

[0046] 1. A pesticide composition comprising:

[0047] (a) at least three compounds selected from the group consisting of soybean oil, eugenol, cinnamaldehyde, tea tree oil, rapeseed oil, clove oil, and spearmint oil as vegetable oil active ingredients; (b) optionally Cu(I) oxide as inorganic active agent;

[0048] (c) a compound selected from the group consisting of xanthan gum, polyvinyl alcohol, magnesium aluminium silicate, ethoxylated vegetable oil, preferably ethoxylated castor oil; benzenesulphonic acid Cl 0-13 alkyl derivative; ethoxylated sorbitan ester; Cl 2- 14 ethoxylated alcohol; potassium polyarylphenyl ether phosphate; sodium salt of methylene polynaphthyl sulphonate as an excipient;

[0049] (d) optionally glycerol as antifreeze;

[0050] (e) optionally an antioxidant, preferably ascorbic acid; and

[0051] (f) optionally water as a carrier, as needed.

[0052] 2. The pesticide composition according to point 1, comprising at least four of the compounds listed in point (a) of point 1 as vegetable oil active ingredients.

[0053] 3. The pesticide composition according to point 1, comprising at least five of the compounds listed in point (a) of point 1 as vegetable oil active ingredients.

[0054] 4. The pesticide composition according to Point 1, comprising the following as active ingredients:

[0055] (a) soybean oil, eugenol, cinnamaldehyde, tea tree oil; or

[0056] (b) soybean oil, eugenol, cinnamaldehyde, tea tree oil, rapeseed oil; or

[0057] (c) clove oil, spearmint oil, tea tree oil; or

[0058] (d) soybean oil, eugenol, cinnamaldehyde, tea tree oil and Cu(I) oxide.

[0059] 5. The composition according to Point 1, comprising clove oil, spearmint oil and tea tree oil as vegetable oil active ingredients.

[0060] 6. The pesticide composition according to Point 1 comprising at least three of the substances listed in point (a) of Point 1 as vegetable oil active ingredients and Cu(I) oxide as inorganic active ingredient.

[0061] 7. A concentrate of a pesticide composition, wherein the pesticide composition according to any one of Points 1 to 6 comprises the following amounts of the vegetable oil or inorganic active ingredients: (a) soybean oil: 100 — 500 g / 1, preferably 200-400 g / 1;

[0062] (b) cinnamaldehyde: 10-40 g / 1, preferably 20-30 g / 1;

[0063] (c) eugenol: 20-100 g / 1, preferably 50 g / 1;

[0064] (d) tea tree oil: 10-150 g / 1, preferably 10-100 g / 1;

[0065] (e) rapeseed oil: 100-500 g / 1, preferably 200-400 g / 1;

[0066] (f) clove oil: 50-200 g / 1, preferably 50-100 g / 1;

[0067] (g) spearmint oil: 50-200 g / 1, preferably 50-100 g / 1;

[0068] (h) Cu(I) oxide: 50-150 g / 1, preferably 50-100 g / 1.

[0069] 8. A concentrate of pesticide composition, wherein the pesticide composition according to Point 4 comprises the following amounts of the active ingredients:

[0070] (a) 220 g / 1 of soybean oil, 50 g / 1 of eugenol, 30 g / 1 of cinnamaldehyde, 10 g / 1 of tea tree oil; or

[0071] (b) 500 g / 1 of soybean oil, 50 g / 1 of eugenol, 20 g / 1 of cinnamaldehyde, 10 g / 1 of tea tree oil, 320 g / 1 of rapeseed oil; or

[0072] (c) 100 g / 1 of clove oil, 100 g / 1 of spearmint oil, 100 g / 1 of tea tree oil; or

[0073] (d) 500 g / 1 of soybean oil, 50 g / 1 of eugenol, 20 g / 1 of cinnamaldehyde, 10 g / 1 of tea tree oil and 100 g / 1 of Cu(I) oxide.

[0074] 9. The pesticide composition or concentrate of pesticide composition according to any one of points 1 to 8 in the form of an aqueous phase oil emulsion, an aqueous phase suspension emulsion or an emulsifiable concentrate.

[0075] 10. Process for the preparation of the pesticide composition according to any one of Points 1 to 9, characterised in that the active ingredients and the excipients, optionally the antifreeze, antioxidant and carrier are mixed by a method known per se, heat treated as required and the product is homogenised and formulated and / or diluted as required.

[0076] 11. The use of the pesticide composition or concentrate of the pesticide composition according to Points 1-10 as a pesticide, wherein the plants to be protected are selected from the group consisting of: apples, pears, cucumbers, tomatoes, leafy vegetables, potatoes, grapes, peppers, strawberries, raspberries, currants, gooseberries, aubergines, beans, apricots, sour cherries, cherries, cereals including wheat, triticale, hops, peaches, plums, sunflowers, grape, oranges, lemons, bananas. 12. The use according to Point 11, wherein the pathogens to be controlled are selected from the group consisting of Botrytis cinerea, Sclerotinia sclerotiorum, Fusarium oxysporum, Fusarium solani, Fusarium sambucinum (F roseum), Monilia laxa, Monilia fructigena, Monilia fructicola, Phytophthora infestans, Fusarium graminearum, Fusarium culmorum, Alternaria sp, Botrytis sp, Fusarium sp, Monilia sp, Penicillium sp.

[0077] 13. The use according to Point 12, wherein the pathogens to be controlled are selected from the group consisting of Fusarium oxysporum, Fusarium solani, Monilia laxa, Monilia fructigena.

[0078] 14. The use according to any one of points 12 to 14, characterized in that prior to the application the pesticide composition or concentrate of pesticide composition is diluted with water to a concentration of at least 0.075 v / v%, preferably 0.125 v / v%, of each of the active ingredient plant oils.

[0079] DETAILED DESCRIPTION OF THE INVENTION

[0080] The subject invention is a pesticide composition comprising:

[0081] (a) a plant oil active ingredient selected from the group consisting of soybean oil, eugenol, cinnamaldehyde, tea tree oil, rapeseed oil, clove oil, and spearmint oil;

[0082] (b) optionally Cu(I) oxide as inorganic active ingredient;

[0083] (c) as excipient, a compound selected from the group consisting of xanthan gum, polyvinyl alcohol, magnesium aluminium silicate, ethoxylated vegetable oil, preferably ethoxylated castor oil; benzenesulphonic acid C10-13 alkyl derivative; ethoxylated sorbitan ester; C12- 14 ethoxylated alcohol; potassium polyarylphenyl ether phosphate; sodium salt of methylene polynaphthyl sulphonate;

[0084] (d) optionally glycerol as antifreeze;

[0085] (e) optionally an antioxidant, preferably ascorbic acid; and

[0086] (f) optionally water as a carrier, as necessary.

[0087] In one embodiment of the invention, the components according to Table 1 are comprised in the amounts indicated therein. Table 1 : Embodiment 1 of the invention

[0088] Type of formulation proposed for the embodiment: EW = oil emulsion in aqueous phase.

[0089] In an embodiment of the invention, the components according to Table 2 are comprised in the amounts indicated therein.

[0090] Table 2: Embodiment 2 of the invention

[0091] Type of formulation proposed for the embodiment: EC = Emulsifiable concentrate.

[0092] In an embodiment of the invention the components according to Table 3 are comprised in the amounts indicated therein. Table 3 : Embodiment 3 of the invention

[0093] Type of formulation proposed for the embodiment: EW = Oil emulsion in aqueous phase. The product contains only organic and / or biodegradable components.

[0094] Recommended crops where the formulation can be used: apples, pears, cucumbers, tomatoes, leafy vegetables, potatoes, grapes, peppers, strawberries, raspberries, aubergines, beans, apricots, sour cherries, cherries, cereals, hops, peaches, plums, sunflowers, rape.

[0095] Suggested uses: open field, greenhouse, post-harvest.

[0096] In an embodiment of the invention, the components according to Table 4 are comprised in the amounts indicated therein.

[0097] Table 4: Embodiment 4 of the invention

[0098] Type of formulation proposed for the embodiment: EW = oil emulsion in aqueous phase. The clove oil, spearmint oil and tea tree oil in this product act together as active ingredients to inhibit the growth of various plant pathogenic fungal species. The product contains organic and / or biodegradable components.

[0099] The plant pathogenic fungal species against which the composition has growth inhibitory activity are Botrytis cinerea. Sclerotinia sclerolionim. Fusarium oxysporum. Fusarium solani. Fusarium sambucinum (F roseum). Monilia laxa. Monilia fructigena, Monilia fructicola, Phytophthora infestans, Fusarium graminearum, Fusarium culmorum, Alternaria sp. Botrytis sp. Fusarium sp. Monilia sp. Penicillium sp.

[0100] Crops in which the product has shown to have a pesticidal effect based on the results of in vitro tests: apples, pears, cucumbers, tomatoes, leafy vegetables, potatoes, grapes, peppers, strawberries, raspberries, currants, gooseberries, aubergines, beans, apricots, sour cherries, cherries, cereals, including wheat, triticale, hops, peaches, plums, sunflowers, rape, oranges, lemons, bananas.

[0101] Suggested uses: open field, greenhouse, post-harvest.

[0102] In an embodiment of the invention, the components according to Table 5 are present in the amounts indicated therein.

[0103] Table 5: Embodiment 5 of the invention

[0104] Type of formulation proposed for the embodiment: SE = water based suspension emulsion

[0105] EXAMPLES

[0106] Example 1: Preparation of the composition according to embodiment 1 of the invention

[0107] To prepare the oil phase, first the soybean oil is measured and we start stirring. Then eugenol, cinnamaldehyde, tea tree oil and Emulsogen EL 360 are added. The mixture is stirred continuously for one hour. In the thickening phase, part of the water is measured and added, stirring is started and then part of the glycerine and xanthan gum are added. The mixture is stirred for two hours. To obtain the final product, the remaining water is measured and mixing is started, then the mixture is heated to 70°C. Ghosenol GL-05 is added and stirred for 30 minutes. Then the remaining glycerol, Van Gel B, the oily phase and the thickener phase are added. The mixture is stirred for 15 minutes with a high shear mixer and finally stirred continuously for 1 hour.

[0108] Example 2: Preparation of the composition according to embodiment 2 of the invention

[0109] The preparation process is carried out as follows: first, the soybean oil is measured and the mixing is started. Then rapeseed oil, eugenol, cinnamaldehyde and tea tree oil are added. Then Nansa EVM 63 / B and finally Emulsogen EL 360 are added.

[0110] Example 3: Preparation of the composition according to embodiment 3 of the invention

[0111] In the thickening phase, a portion of the water is measured, stirring is started, then a portion of the glycerol and xanthan gum are added and stirred for two hours. To obtain the final product, the remaining water is measured, the stirring is started and the mixture is heated to 50°C. Ghosenol GL-05 is added and stirred for 30 minutes. Then the ascorbic acid, the rest of the glycerol, Van Gel B, Tween 20, clove oil, peppermint oil and tea tree oil are added. Finally, the thickening phase is added and the mixture is stirred with a high shear mixer for 15 minutes and then stirred continuously for 1 hour. Example 4: Preparation of the composition according to embodiment 4 of the invention

[0112] During the thickening phase, a portion of the water is measured, stirring is started, then a portion of the glycerol and Rhodopol 23 are added and stirred for two hours. To obtain the final product, the remaining water is measured, stirring is started and the mixture is heated to 50°C. Kuraray Poval is added and stirred for 30 minutes. Then ascorbic acid, the rest of the glycerol, Van Gel B, Alkamuls TS20, clove oil, spearmint oil and tea tree oil are added. Finally, the thickening phase is added, stirred with a high shear mixer for 15 minutes and then stirred continuously for 1 hour.

[0113] Example 5: Preparation of the composition according to embodiment 5 of the invention

[0114] During the thickening phase, a portion of the water is measured, the stirring is started, then the xanthan gum is added and stirred for two hours. To prepare the oil phase, the soybean oil is measured and stirring is started, then eugenol, cinnamon dehyde, tea tree oil, Nansa EVM 63 / B and Rhodasurf LA 30 are added. This mixture is stirred for 1 hour. To obtain the final product, the remaining water is measured, stirring is started then Soprophor FLK, Supragil RM 210 El and Cu(I) oxide are added. The mixture is stirred for 30 minutes with a high shear mixer, then the oily phase is added and stirred again for 30 minutes with a high shear mixer. Then the thickener phase is added, stirred for 15 minutes with a high shear mixer then stirred continuously for 1 hour.

[0115] EFFICACY STUDIES

[0116] The efficacy of the composition according to the invention is demonstrated by the following test results.

[0117] The efficacy results of embodiment 4 of the invention are summarised below.

[0118] 1. Test results of the fungicidal activity of the composition according to the invention

[0119] Duration of the tests: 16.04.2024-05.09.2024.

[0120] Location of the tests: MATE Georgikon Campus, Institute of Plant Protection, Department of Plant Protection, Laboratories of the Imre Festetics Bioinnovation Centre 8360 Keszthely Festetics u. 7.

[0121] The materials and methods of the studies: the study of the fungicide activity of the composition according to embodiment 4 of the invention was carried out on infected potato dextrose agar plates in single-use Petri dishes. In the experiment, the composition of embodiment 4 was used at dilutions of 0.125, 0.250; 0.500; 0.750 and 1.125 v / v%. The facultative parasitic phytopathogenic fungi used in the studies were Alternaria sp., Botrytis sp., Fusarium sp., Monilia sp., Penicillium sp. The control fungal cultures were grown on potato dextrose agar without the composition of embodiment 4. The experiment was set up in 5-5 replicates per pathogen and dilution. After inoculation, incubation was at 22°C. Evaluation was performed by visual examination on days 5, 7, 11 and 21 after the pathogen inoculation.

[0122] Test results: No mycelial growth was observed in Petri dishes containing the composition of embodiment 4 for any of the fungal species tested at all the applied concentrations on days 5, 7, 11 and 21 post-inoculation. Accordingly, a total fungicidal effect was observed in the treated Petri dishes. The results are quantified in the tables below:

[0123] Table 6. Results (Keszthely, 22 Apr, day 5 after inoculation)

[0124] Table 7. Results (Keszthely, 24 Apr, day 7 after inoculation) Table 8. Results (Keszthely, 29 Apr. day 11 after inoculation)

[0125] Table 9. Results (Keszthely, 9 May, day 21 after inoculation)

[0126] From the above, the surprising and outstanding fungicidal activity of the composition according to the invention can be concluded.

[0127] Further studies were carried out with embodiment 4 of the invention to test the antifungal activity of the compositions of the invention in vitro.

[0128] Isolates and methods used in the experiment

[0129] In the laboratory study, the growth activity of one isolate of each of nine plant pathogenic fungal species of the EW formulation according to embodiment 4 containing three different plant essential oils was evaluated using the agar plate method. The fungal species included in the experiment were Botrytis cinerea, Sclerotinia sclerotiorum, Fusarium oxysporum, Fusarium solani, Fusarium sambucinum (F roseum), Monilia laxa, Monilia fructigena, Monilia fructicola, Phytophthora infe stans.

[0130] The experiments were set up in 4 replicates, on media corresponding to the individual nutrient requirements of the species. The antifungal activity of the vegetable oil mixture was tested at five different concentrations and an untreated control treatment was also included. The concentrations (v / v%) used for the in vitro study were 0.125%, 0.25%, 0.5%, 0.75%, 1.125%. To determine the in vitro efficacy of the essential oil mixture, the diameter of fungal colonies was measured in the cultures after 72, 120 and 168 hours.

[0131] LABORATORY EXPERIMENTAL RESULTS

[0132] Figure 9 shows the in vitro efficacy of the essential oil mixture against Botrytis cinerea. The essential oil mixture proved complete mycelial growth inhibition of Botrytis cinerea even when applied at the lowest concentration of 0.125% tested.

[0133] Figure 13 shows the in vitro efficacy of the essential oil mixture against the pathogen Sclerotinia sclerotiorum. The essential oil mixture significantly inhibited the growth of the pathogen at a concentration of 0.25 %. At 0.125%, the lowest concentration tested, the inhibitory effect observed after 72 hours was not persistent, and after further 96 hours the growth vigour of the fungal strain was similar to that of the control culture.

[0134] Figure 10 shows the in vitro efficacy of the essential oil mixture against the pathogen Fusarium oxysporum. The essential oil mixture, applied at the lowest concentration of 0.125% tested, significantly reduced the growth of the Fusarium oxysporum pathogen by about one third. Total mycelial growth inhibition was achieved at 0.25%.

[0135] Figure 11 shows the in vitro efficacy of the essential oil mixture against the pathogen Fusarium solani. There was a significant difference between the first three treatment levels when comparing each time. On the medium containing the mixture at a concentration of 1.125%, Fusarium solani did not start to develop even after 7 days, demonstrating complete inhibition.

[0136] Figure 12 shows the in vitro efficacy of the essential oil mixture against Fusarium sambucinum. At the highest concentration of the essential oil mixture tested, the growth inhibition effect against Fusarium sambucinum is significant but not complete.

[0137] Figure 7 shows the in vitro efficacy of the essential oil mixture against Monilia fructicola. On media containing the essential oil mixture at a concentration of 0.25% or higher, the pathogen Monilia fructicola did not start to grow by the time of evaluation and thus complete inhibition was observed.

[0138] Figure 8 shows the in vitro efficacy of the essential oil mixture against Monilia fructigena. Cultures inoculated on medium containing different concentrations of essential oils did not start to grow even after five days, thus complete inhibition was observed at the concentrations tested.

[0139] Figure 6 shows the in vitro efficacy of the essential oil mixture against Monilia laxa. The lower concentrations used during the treatment significantly inhibited the growth of Monilia laxa cultures, while concentrations of 0.5 % and higher inhibited it completely.

[0140] Summary of the results

[0141] The series of tests carried out under in vitro conditions demonstrated that the tested essential oil mixture was effective against all plant pathogens in the experiment. The extent of growth inhibition after 72 hours compared to the control is illustrated in the table below:

[0142] Table 10. Degree of growth inhibition compared to control

[0143] The essential oil mixture, applied at 0.125%, the lowest concentration tested, demonstrated complete mycelial growth inhibition on Botrytis cinerea and Monilia fructigena isolates after 7 days.

[0144] For the Fusarium oxysporum and Monilia fructicola isolates, growth inhibition was complete for media containing essential oil mixtures of 0.25% and higher concentrations.

[0145] Complete growth inhibition of Monilia laxa isolates was observed at 0.5% concentration.

[0146] On the medium containing 1.125% essential oil mixture, the pathogen Fusarium solani did not grow even after 7 days and complete inhibition was observed.

[0147] In the case of Sclerotinia sclerotiorum and Fusarium sambucinum (F roseum). the highest concentration tested (1.125%) had a strong, significant inhibitory effect on the growth of the pathogen, which effect was not complete after 7 days.

[0148] Due to the weak growth vigour of Phytophthora infestans, the study takes longer time.

[0149] In possession of the above results, further efficacy studies were conducted including three additional pathogens.

[0150] Isolates and methods used in the studies

[0151] In the laboratory study, the individual effect of three EW type preparations of three different plant essential oils of embodiment 4 on the growth activity of three isolates of three plant pathogenic fungal species was evaluated by the agar plate method. The fungal species included in the experiment are Phytophthora infestans, Fusarium graminearum, Fusarium culmorum. The experiments were set up in 4 replicates, on media corresponding to the individual nutrient requirements of the species. The antifungal activity of the vegetable oil mixture was tested at five different concentrations and an untreated control treatment was also included. The concentrations (v / v%) used for the in vitro study were 0.125%, 0.25%, 0.5%, 0.75%, 1.125%. To determine the in vitro efficacy of the essential oil mixture, the diameter of fungal colonies was measured in the cultures after 72, 120 and 168 hours.

[0152] LABORATORY EXPERIMENTAL RESULTS

[0153] Figure 5 shows the in vitro efficacy of the essential oil mixture against Phytophthora infestans. The growth vigour of this fungal strain is weak in specific culture media of different compositions. When the in vitro efficacy of the essential oil mixture was tested on media containing different concentrations of the mixture, the cultures did not grow even after three weeks. The essential oil mixture demonstrated complete mycelial growth inhibition of the pathogen Phytophthora infestans even when applied at 0.125%, the lowest concentration tested.

[0154] Figure 3 shows the in vitro efficacy of the essential oil mixture against the pathogen Fusarium graminearum. The essential oil mixture significantly suppressed the growth of the pathogen Fusarium graminearum even when applied at 0.125%, the lowest concentration tested. Total mycelial growth inhibition was achieved at the concentration of 0.25%.

[0155] Figure 4 shows the in vitro efficacy of the essential oil mixture against the pathogen Fusarium culmorum. Concentrations of 0.25% and higher applied during the treatment completely inhibited the growth of Fusarium culmorum cultures.

[0156] Summary of results

[0157] A series of tests carried out under in vitro conditions demonstrated that the tested essential oil mixture was effective against all plant pathogens in the experiment. The extent of growth inhibition after 72 hours compared to the control is illustrated in the table below.

[0158] Table 11. Degree of growth inhibition compared to control

[0159] In case of the Phytophthora infestans isolate with poor growth vigour, the essential oil mixture at 0.125%, the lowest concentration tested, already demonstrated complete mycelial growth inhibition.

[0160] In case of the Fusarium graminearum and Fusarium culmorum isolates, growth inhibition was complete for media containing the essential oil mixture at concentrations of 0.25% and higher.

[0161] A composition containing the three plant oil active ingredients of the invention was further tested. During the tests, it was examined whether the composition according to the present invention showed a technical added effect compared to the state of the art fcompositions only comprising only any one or two vegetable oil active ingredients of the three vegetable oil active ingredients according to the invention.

[0162] Isolates and methods used during the study

[0163] During the laboratory test, the independent effect of seven EW-type preparations (essential oils and their mixtures) on the growth of isolates of four plant pathogenic fungal species was investigated using the agar plate method.

[0164] The essential oils and their mixtures used in the tests:

[0165] - spearmint oil EW

[0166] - clove oil EW

[0167] - cinnamaldehyde EW

[0168] - spearmint oil + clove oil EW

[0169] - spearmint oil + cinnamaldehyde EW

[0170] - clove oil + cinnamaldehyde EW

[0171] - clove oil + spearmint oil + tea tree oil EW List of fungi species used in the tests:

[0172] • Monilinia laxa

[0173] • Monilinia fructigena

[0174] • Fusarium oxysporum

[0175] • Fusarium solani

[0176] The tests were set up in 4 replicates, on media corresponding to the individual nutrient requirements of the species. The antifungal activity of the vegetable oils and their mixtures was tested at three different concentrations, and an untreated control treatment was also included.

[0177] Concentrations used in the in vitro study (v / v%):

[0178] - 0.005%

[0179] - 0.075%

[0180] - 0.125%

[0181] To determine the in vitro efficacy of the essential oils and their mixtures, the diameter of fungal colonies was measured in the cultures on days 3, 5 and 7 after inoculation.

[0182] LABORATORY EXPERIMENTAL RESULTS

[0183] Figure 15 shows the in vitro efficacy of the essential oils and their mixtures against Monilinia laxa. Of the essential oil components, cinnamaldehyde EW has the weakest growth inhibition effect on Monilinia laxa. The efficacy observed for the spearmint EW was relatively higher, with an inhibitory effect above 20% in all three concentrations. The effectiveness of clove oil was 26.5% at the highest concentration tested.

[0184] The cumulative effect of using different essential oils together was not significant in case of two components, with the highest inhibitory effect (27.6%) observed for spearmint+clove oil EW.

[0185] The three-component mixture resulted in an inhibitory effect of 64-83% at the two highest concentrations tested. The efficacy for this pathogen was also above 20% at lower concentrations.

[0186] Figure 16 shows the in vitro efficacy of essential oils and their mixtures against the pathogen Monilinia fructigena.

[0187] Among the essential oils tested, cinnamaldehyde EW has the lowest efficacy against Monilinia fructigena. axa. At the highest tested concentration of 0.125 v / v%, spearmint oil and clove oil showed efficacies of 38 and 39%, respectively. The combined use of spearmint oil and cinnamaldehyde in all three concentrations increased the inhibitory effect of the individual components. For the clove oil + cinnamaldehyde mixture, no cumulative effect was observed at the two higher concentrations. The spearmint + clove oil EW mixture also increased the efficacy of the essential oils alone at the two higher concentrations tested. The latter essential oil mixture demonstrated a significant mycelium growth inhibition of 44.7% at a concentration of 0.125v / v%.

[0188] The triple essential oil mixture proved to be the most effective against the Monilinia fructigena isolate at all three concentrations tested. Even at a concentration of 0.075%, the clove oil + spearmint oil + tea tree oil EW formulation reduced the size of fungal colonies by more than three quarters.

[0189] Of the pathogens tested, this fungal species proved to be the most sensitive to the essential oils tested.

[0190] Figure 17 shows the in vitro efficacy of essential oils and their mixtures against the pathogen Fusarium oxysporum.

[0191] Among the single component products, spearmint EW had the weakest inhibitory effect on the growth of fungal colonies compared to the control colonies based on the average of the concentrations tested. In case of clove oil EW, this inhibitory effect is almost twice as high, 8.1%.

[0192] In comparing the two-component mixtures, the best result was obtained with the clove oil + cinnamaldehyde EW, with a maximum inhibition value of 16%.

[0193] Against the Fusarium oxysporum isolate at concentrations of 0.075 v / v% and 0.125 v / v%, the clove oil + spearmint oil + tea tree oil mixture proved to be the most effective. The three-component formulation including tea tree oil exceeded the average efficacy of the other formulations more than four times, but the inhibitory effect was still only 4.2% at the lowest concentration (0.005 v / v%).

[0194] Figure 18 shows the in vitro efficacy of essential oils and their mixtures against the pathogen Fusarium solani.

[0195] Of all the essential oils tested, spearmint oil showed the lowest efficacy against Fusarium solani isolates, too, at all three concentrations tested. Furthermore, it is proportionally even more inferior to the efficacy of the other two single-ingredient preparations.

[0196] In the case of the two-component formulations, clove oil + cinnamaldehyde EW was also the most effective at the highest concentration tested, but in terms of averages, spearmint oil + clove oil EW proved to be more effective. The performance of the three-component formulation was also outstanding here, but it did not stand out as much in terms of proportions as it was seen with the Fusarium oxysporum isolate. The highest inhibition rate was 40.1 %.

[0197] Summary of the results

[0198] The percentage of growth inhibition compared to control cultures is shown in Table 12 below:

[0199] A series of tests carried out under in vitro conditions demonstrated that the tested essential oils and their mixtures can inhibit the development of all plant pathogens in the experiment.

[0200] The cinnamaldehyde and clove oil showed a more stable effect, thus proving more effective against Fusarium pathogens than spearmint oil when used in higher concentrations.

[0201] In this series of tests, the mixture of spearmint oil + clove oil was found to be the most effective of the two-component products. The maximum value of the inhibitory effects against Fusarium species was 18.4% in case of the application of the clove oil + cinnamaldehyde mixture. The representatives of the genus Monilinia showed a higher susceptibility to the two- component mixtures, with 27.6% inhibition for Monilinia laxa and 44.7% for Monilinia fructigena isolates when using the mixture of spearmint oil + clove oil.

[0202] The three-component composition of clove oil + spearmint oil + tea tree oil EW significantly inhibited the growth of pathogens at concentrations of 0.075 and 0.125 v / v%, the inhibition was 60.6% in average. Compared to the efficacy of the spearmint oil + clove oil EW, the preparation also comprising the tea tree oil component showed a significant, 2.5-fold higher efficacy against Fusarium and Monilinia pathogens in this series of tests.

[0203] Photo documentation of the experimental series is shown in Figure 19.

[0204] In conclusion, the anti-pathogenic activity of the compositions comprising three vegetable oil active ingredients is unexpectedly and significantly superior to that of the state of the art compositions comprising one or two components.

[0205] INDUSTRIAL APPLICABILITY

[0206] In the light of the above, the composition according to the invention is suitable as a natural pesticide product for plant protection. The composition according to the present invention is suitable for use in the industry, especially in the field of pesticide production.

Claims

Claims1. A pesticide composition comprising:(g) at least three compounds selected from the group consisting of soybean oil, eugenol, cinnamaldehyde, tea tree oil, rapeseed oil, clove oil, and spearmint oil as vegetable oil active ingredients;(h) optionally Cu(I) oxide as inorganic active agent;(i) a compound selected from the group consisting of xanthan gum, polyvinyl alcohol, magnesium aluminium silicate, ethoxylated vegetable oil, preferably ethoxylated castor oil; benzenesulphonic acid CIO-13 alkyl derivative; ethoxylated sorbitan ester; C12-14 ethoxylated alcohol; potassium polyarylphenyl ether phosphate; sodium salt of methylene polynaphthyl sulphonate as an excipient;(j) optionally glycerol as antifreeze;(k) optionally an antioxidant, preferably ascorbic acid; and(l) optionally water as a carrier, as needed.

2. The pesticide composition according to Claim 1, comprising at least four of the compounds listed in point (a) of Claim 1 as vegetable oil active ingredients.

3. The pesticide composition according to Claim 1, comprising at least five of the compounds listed in point (a) of Claim 1 as vegetable oil active ingredients.

4. The pesticide composition according to Claim 1, comprising the following as active ingredients:(e) soybean oil, eugenol, cinnamaldehyde, tea tree oil; or(f) soybean oil, eugenol, cinnamaldehyde, tea tree oil, rapeseed oil; or(g) clove oil, spearmint oil, tea tree oil; or(h) soybean oil, eugenol, cinnamaldehyde, tea tree oil and Cu(I) oxide.

5. The composition according to Claim 1, comprising clove oil, spearmint oil and tea tree oil as vegetable oil active ingredients.

6. The pesticide composition according to Claim 1 comprising at least three of the substances listed in point (a) of Claim 1 as vegetable oil active ingredients and Cu(I) oxide as inorganic active ingredient.

7. A concentrate of a pesticide composition, wherein the pesticide composition according to any one of Claims 1 to 6 comprises the following amounts of the vegetable oil or inorganic active ingredients:(h) soybean oil: 100-500 g / 1, preferably 200-400 g / 1;(i) cinnamaldehyde: 10-40 g / 1, preferably 20-30 g / 1;(j) eugenol: 20-100 g / 1, preferably 50 g / 1;(k) tea tree oil: 10-150 g / 1, preferably 10-100 g / 1;(l) rapeseed oil: 100-500 g / 1, preferably 200-400 g / 1;(m) clove oil: 50-200 g / 1, preferably 50-100 g / 1;(n) spearmint oil: 50-200 g / 1, preferably 50-100 g / 1;(h) Cu(I) oxide: 50-150 g / 1, preferably 50-100 g / 1.

8. A concentrate of pesticide composition, wherein the pesticide composition according to Claim 4 comprises the following amounts of the active ingredients:(a) 220 g / 1 of soybean oil, 50 g / 1 of eugenol, 30 g / 1 of cinnamaldehyde, 10 g / 1 of tea tree oil; or(b) 500 g / 1 of soybean oil, 50 g / 1 of eugenol, 20 g / 1 of cinnamaldehyde, 10 g / 1 of tea tree oil, 320 g / 1 of rapeseed oil; or(e) 100 g / 1 of clove oil, 100 g / 1 of spearmint oil, 100 g / 1 of tea tree oil; or(f) 500 g / 1 of soybean oil, 50 g / 1 of eugenol, 20 g / 1 of cinnamaldehyde, 10 g / 1 of tea tree oil and 100 g / 1 of Cu(I) oxide.

9. The pesticide composition or concentrate of pesticide composition according to any one of claims 1 to 8 in the form of an aqueous phase oil emulsion, an aqueous phase suspension emulsion or an emulsifiable concentrate.

10. Process for the preparation of the pesticide composition according to any one of Claims 1 to 9, characterised in that the active ingredients and the excipients, optionally the antifreeze, antioxidant and carrier are mixed by a method known per se, heat treated as required and the product is homogenised and formulated and / or diluted as required.

11. The use of the pesticide composition or concentrate of the pesticide composition according to Claims 1-10 as a pesticide, wherein the plants to be protected are selected from the group consisting of: apples, pears, cucumbers, tomatoes, leafy vegetables, potatoes, grapes, peppers, strawberries, raspberries, currants, gooseberries, aubergines, beans, apricots, sour cherries, cherries, cereals including wheat, triticale, hops, peaches, plums, sunflowers, rape, oranges, lemons, bananas.

12. The use according to Claim 11, wherein the pathogens to be controlled are selected from the group consisting of Botrytis cinerea, Sclerotinia sclerotiorum, Fusarium oxysporum, Fusarium solani, Fusarium sambucinum (F roseum), Monilia laxa, Monilia fructigena, Monilia fructicola, Phytophthora infestans, Fusarium graminearum, Fusarium culmorum, Alternaria sp, Botrytis sp, Fusarium sp, Monilia sp, Penicillium sp.

13. The use according to Claim 12, wherein the pathogens to be controlled are selected from the group consisting of Fusarium oxysporum, Fusarium solani, Monilia laxa, Monilia fructigena.

14. The use according to any one of claims 12 to 13, characterized in that prior to the application the pesticide composition or concentrate of pesticide composition is diluted with water to a concentration of at least 0.075 v / v%, preferably 0.125 v / v%, of each of the active ingredient plant oils.

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