Cardenolide extracts

Abstract

The present invention relates to the field of protecting seeds, such as grains or any another part of a plant capable of forming a complete plant, from attacks by birds, and to the treatment thereof using compounds of natural origin.

Description

[0001] CARDENOLIDES EXTRACTS

[0002] The present invention relates to the field of protecting seeds, such as grains or other parts of a plant capable of forming a complete plant, against attacks by birds and to the treatment of these with compounds of natural origin.

[0003] In agriculture, farmers sow seeds such as corn, sunflower, soybean, peas, and other cereals in their fields within furrows made in the soil. However, birds, such as corvids, cause significant damage to the seeds that have been sown, which results in significant economic losses for the farmer. And even when we try to camouflage the furrows in which the seeds have been sown, we observe that birds still manage to find them to eat them.

[0004] Faced with this risk, which is difficult to predict and control, there is a need to take certain precautions when planting and maintaining crops.

[0005] Seed treatments are commercially available for bird repellent use. These chemicals contain active substances from the carbamate family (thiram or ziram), which have been the subject of experimental testing for several years but are not recommended due to environmental concerns.

[0006] The work of Brower and Fink (Ann NY Acad Sci. 1985;443:171-88) first reported that the presence of cardenolides in certain butterfly species (page 173, 1st paragraph) had an emetic effect on birds. This effect is due to the presence of cardenolides accumulated by ingestion of Asclepias leaves ("milkweed" in English). Thus, cardenolides present in plants of the genus Asclepias are known for their emetic effect on birds.

[0007] Ground materials from plants of the genus Asclepias were proposed in application WO 2020 / 109529, to protect seeds from attacks by birds and prevent them from being eaten by them. The products used are raw and complete ground materials from an Asclepias plant, in particular Asclepias leaves. The ground material is used as is and does not undergo any processing or extraction after grinding. The raw ground material is used to form a film around seeds to be protected against birds and for the production of plant protection products.

[0008] There is a need to develop extracts and formulations suitable for coating seeds to protect them from bird attacks.

[0009] In this context, the invention aims to provide a method for producing new extracts of at least one plant of the genus Asclepias, which are concentrated in cardenolides. In particular, the invention provides methods that are easy to implement industrially. The invention thus provides new extracts that are particularly suitable for use as bird repellents, and therefore for ensuring seed protection. The present invention thus relates to an extract of at least one plant containing cardenolides, characterized in that it comprises certain proportions of cardenolides and dry matter.

[0010] Such an extract according to the invention has the performance advantages for bird repulsion and a simplification in its use for seed coating while allowing control of the doses applied to the seeds.

[0011] Such an extract is particularly suitable for protecting seeds from attacks by birds, which eat the seeds after sowing or at the time of germination. In particular, this effect is observed against corvids, especially crows and ravens, especially rooks and other bird pests.

[0012] The extract according to the invention also has industrial advantages. A liquid extract firstly allows for more precise dosing. The extract can also be concentrated, thus allowing for similar effectiveness but with a smaller application volume on the seeds. This implies a reduction in the problems of fouling of industrial coating machines, an improvement in throughput (drying time or penetration of the applied mixtures extended if the volumes are too high), as well as a gain in quality if the stickiness of the treatment resulting from an excessively large applied volume leads to caking in the bags where the seeds are stored.

[0013] Finally, the application of too much “dry” material in seed treatment, which has no binding character but is powdery, can lead to the generation of dust, an essential quality characteristic which is subject to strict regulations.

[0014] The invention thus relates to extracts, preferably extracts of at least one plant containing cardenolides, comprising at least:

[0015] - 0.5 to 4% by weight of cardenolides, relative to the total weight of said extract,

[0016] - 10 to 85% by weight of dry matter, relative to the total weight of said extract,

[0017] - 1 to 8% by weight of cardenolides, relative to the total weight of said dry matter, with said extract having a viscosity of between 1 and 10000 mPa.s.

[0018] The present invention also relates to methods for obtaining extracts of at least one plant containing cardenolides, as well as to formulations comprising such extracts, seeds coated with extracts, and the use of these extracts and formulations for treating seeds, in particular as a bird repellent, in particular to prevent birds from consuming seeds coated with said extracts. Definitions

[0019] Seeds are a part of a plant, such as a seed, kernel or pip, capable of forming a complete plant and which can be sown.

[0020] Plants are defined as photosynthetic and autotrophic organisms characterized by plant cells. The term "plant" as used herein refers to a member of the Kingdom Plantae and includes all stages of the plant life cycle, including, without limitation, seeds, and includes all parts of the plant. Plants according to the present invention may be agricultural and horticultural plants, shrubs, trees, and herbs, hereinafter sometimes collectively referred to as plants.

[0021] Cardenolides are molecules of the cardanolide subtype, which are themselves a class of steroids, including cardioactive steroids.

[0022] Dry matter is the material comprising mineral and organic matter that is obtained when water is removed from a product. Dry matter quantifies the total biomass of a crop produced on a dry weight basis.

[0023] Throughout this description, unless the context otherwise requires, the word "comprise" or "include", or variations thereof, shall be understood to imply the inclusion of a stated element or method step or group of elements or method steps, but not the exclusion of any other element or method step or group of elements or method steps. The words "comprise" and "include", and variations thereof, may also include the notion of "consisting exclusively of".

[0024] As used in this description, the singular forms "a," "an," and "the" include plural aspects unless the context clearly dictates otherwise.

[0025] The term “and / or” includes the meanings “and”, “or” and all other possible combinations of the elements related to this term.

[0026] The term "between" should be understood to include boundaries.

[0027] Ratios, concentrations, quantities, and other numerical data may be presented in a range format. It should be understood that such a range format is used solely for convenience and brevity and should be interpreted flexibly to include not only the numerical values ​​explicitly stated as limits of the range, but also to include all individual numerical values, or subranges encompassed within that range as if each numerical value and subrange were explicitly stated. Excerpts

[0028] The invention thus relates to an extract, preferably an extract of at least one plant containing cardenolides, comprising at least:

[0029] - 0.5 to 4% by weight of cardenolides, relative to the total weight of said extract,

[0030] - 10 to 85% by weight of dry matter, relative to the total weight of said extract,

[0031] - 1 to 8% by weight of cardenolides, relative to the total weight of said dry matter, with said extract having a viscosity of between 1 and 10000 mPa.s.

[0032] This extract may comprise from 0.5 to 4% by weight of cardenolides, for example from 0.7 to 4% by weight of cardenolides, for example from 1 to 4% by weight of cardenolides, relative to the total weight of said extract.

[0033] This extract may comprise at least 25% by weight of dry matter, for example at least 50% by weight of dry matter, relative to the total weight of said extract. This extract may comprise up to 80% by weight of dry matter, for example up to 70% by weight of dry matter, relative to the total weight of said extract. In particular, the extract comprises from 10 to 85% by weight of dry matter, preferably from 25 to 85% by weight of dry matter, relative to the total weight of said extract.

[0034] This extract may comprise from 1.25 to 4.75% by weight of cardenolides, for example from 1.5 to 4.5% by weight, in particular from 1.5 to 3.5% by weight, particularly from 1.5 to 2% by weight, relative to the total weight of its dry matter.

[0035] Preferably, the extract includes at least:

[0036] - 1.5 to 3.5% by weight of cardenolides, relative to the total weight of said extract, and

[0037] - 10 to 70% by weight of dry matter, relative to the total weight of said extract.

[0038] The extract according to the invention is not, advantageously, a crude ground product of Asclepias or Asclepias leaves.

[0039] This extract may comprise a quantity by weight of cardenolides, relative to the total weight of said extract, greater by at least 10%, for example at least 20%, for example at least 50%, relative to the quantity by weight of cardenolides in said plant.

[0040] Cardenolides generally have a structure that incorporates the following motif (I) with different possible substitutions.

[0041] According to the invention, the cardenolides may be chosen from the group consisting of: calotoxin, uscharidin, asclepin, calactin, calotropagenin, coroglaucigenin, xysmalogenin, voruscharin, gofruside, uzarigenin, 12p-hydroxycoroglaucigenin, 12p~ hydroxycalotropin, 6'-O-feruloyl-desglucouzarin, 12p, 14p-dihydroxy-3p, 19-epoxy-3a-methoxy-5a-card-20(22)-enolide, xysmalogenin, coroglaucigenin, 6-O-(E-4-hydroxycinnamoyl)-desglucouzarin 6-O-sinapinoyl-desglucouzarin, calactin, 16a- acetoxy calactin, calotropin 16a-acetoxycalotropin, 16a-hydroxyasclepin, 16a-acetoxyasclepin uscharin, 16a-hydroxyuscharin, 19-nor-16a-acetoxy-10p-hydroxyasclepin, desglucouzarin, 6'-O-feruloyl-desglucouzarin, and calotropin. One or more types of cardenolides may be used together.

[0042] Preferably, the cardenolides are chosen from calotoxin, uscharidin, asclepin and calactin.

[0043] Examples of typical cardenolides contained in plants of the genus Asclepias include, in particular, calotoxin, uscharidin, asclepin and calactin.

[0044] Dry matter may include lignocellulosic compounds, sugars, salts, polyphenols, cardenolides, organic acids, and proteins.

[0045] The viscosity of the extract can be measured as follows: in a 600 mL beaker (height 12.4 cm and diameter 9 cm), 300 g of extract or formulation are weighed and a Brookfield viscometer type RVDV at 20 rpm (revolutions per minute) with a needle is used to obtain a reliable measurement. Once the stabilization plateau is reached, the value expressed in mPa.s is noted. The temperature must be 25 °C + / - 1 °C or 30 °C + / - 1 °C.

[0046] Process for obtaining an extract from at least one plant

[0047] The invention also relates to a method for obtaining an extract of at least one plant containing cardenolides, comprising at least the following successive steps: a) bringing a raw material of at least one plant containing cardenolides into contact, in a cardenolide extraction solvent S, b) optionally separating the result of the contacting of step a), to recover its liquid part L1, c) carrying out a selective precipitation of the constituents other than the cardenolides of the mixture obtained in step a) or of the liquid part L1 obtained in step b), d) carrying out one or more separations of the result of step c) to recover its liquid part L2, and e) carrying out a concentration of the liquid part L2 to recover an extract concentrated in cardenolides.

[0048] Plants containing cardenolides are preferentially selected among: the apocynacae: Asclepias curassavica, Asclepias eriocarpa, Asclepias incarnate, Asclepias labriformis, Asclepias syriaca, Asclepias tuberosa, Asclepias fruticosa, Cryptostegia, Oxlosteurama, Oxloystera Pergularia tomentosa, Periploca graeca, Periploca nigrescens, Periploca forrestii, Calotropis procera, Calotropis gigantea, et les brassicaceae: Erysimum cheiri, Erysimum canescens, et Erysimum cheiranthoides.

[0049] Preferably, the plant extracts produced within the scope of the invention can be derived from any type of plants of the genus Asclepias, and in particular from plants of one of the species A. (Asclepias) curassavica, A. tuberosa, A. asperula, A. incamata, A. incamata, A. fruosa. viridis, A. speciosa, A. verticillata or A. sullivanti. The A. curassavica species is particularly preferred, in the context of the invention.

[0050] The plants used for the production of an extract according to the invention can, in a conventional manner, be cultivated under standardized cultivation conditions and in a controlled environment. This facilitates the reproducibility and optimization of the process according to the invention.

[0051] The raw material of at least one plant containing cardenolides preferably corresponds to the leaves of at least one plant, to a set of leaves and stems of at least one plant, to the complete aerial parts of at least one plant, or to at least one whole plant. To improve extraction, the whole plant or parts of the plant may be ground beforehand. Before grinding, the plant is preferably dried. Before or after grinding, the raw material may be subjected to dehydration, or even to freeze-drying. Dehydration is carried out using any suitable device well known to those skilled in the art, such as a hot air flow device, a cylinder dryer, a zeolite device or a freeze-dryer, in the case of freeze-drying.

[0052] Step a) thus advantageously consists of bringing into contact a raw material of at least one plant containing cardenolides, in a cardenolide extraction solvent S. Step a) can be carried out in different ways, such as for example by maceration or extractions, in particular by Soxhlet extraction or continuous counter-current extraction for example.

[0053] For maceration, the whole plant or certain parts of the plant are brought into contact with at least one cardenolide extraction solvent S. One or more solvents may be used. According to the invention, a cardenolide extraction solvent S refers to a solvent capable of extracting, in whole or in part, preferably at least one or more types of cardenolides from a raw material of at least one plant containing cardenolides. The solvent used may be any polar solvent, for example water, ethanol, methanol, chloroform, diethyl ether, dichloromethane, ethyl acetate, DMSO. Aqueous solutions or water / organic solvent mixtures may be considered. Several polar solvents may be mixed in various proportions, for example a water / ethanol or water / methanol mixture. The extraction solvent S is preferably water or an aqueous solution.Such solvents make it possible to solubilize the cardenolides of interest, and, in particular, calotoxin, uscharidin, asclepin and calactin.

[0054] The extraction solvent may, in particular, be an aqueous solution containing an alcoholic solvent, such as methanol or ethanol, preferably present at a level of less than 20% v / v.

[0055] For practicality on an industrial scale, the extraction solvent is preferably water or an aqueous solution consisting of water and a buffer such as a phosphate.

[0056] Several dilution levels can be used; for example, 5-12 g of water can be used to extract 1 g of biomass. The contact is preferably carried out at a temperature lower than or equal to the boiling point of the solvent, in particular between 20 and 95 °C. The use of such a temperature makes it possible in particular to minimize the risks of degradation, and in particular of hydrolysis, of the cardenolides of interest. Thus, in general, all the steps of the process according to the invention are preferably carried out at a temperature lower than 85 °C, and preferably lower than 60 °C.

[0057] Maceration can last between 30 minutes and 3 hours.

[0058] The biomass can be extracted several times by renewing the solvent in order to recover as many cardenolides as possible.

[0059] Advantageously, the maceration is carried out with a ratio of mass of plant raw material in grams / volume of extraction solvent in liters of 66.7 to 300, preferably 100 to 200.

[0060] Step b), optional, consists of carrying out a separation of the result of the contacting of step a), to recover its liquid part LL

[0061] This separation can be done using separation methods, for example by filtration, centrifugation or decantation. Separation by filtration is preferred at this stage. In particular, the result of the maceration can be routed to pass through a filter, which will retain the plant material. Thus, the liquid part Ll, which is already rich in cardenolides, is recovered.

[0062] For example, a centrifugal decanter, type Flottweg Z4D4 can be used. The mixture enters the decanter with a dry matter content, for example, of between 7 and 17% by weight. After decantation, a liquid phase L1 is recovered with a dry matter content preferably of between 1 and 5% by weight, in particular between 2 and 4% by weight. For example, by filtration, the mixture arrives on a paper filter preferably having pores of 5-50 μm; the juice L1 can be vacuumed and the wet biomass remains on the filter. The liquid phase L1 can have a dry matter content of between 1 and 5% by weight, in particular between 2 and 4% by weight.

[0063] Advantageously, step c) consists of carrying out a selective precipitation of the constituents other than the cardenolides of the mixture obtained in step a) or of the liquid part L1 obtained in step b).

[0064] Such a step can be carried out in different ways, such as by flocculation of solid materials. Advantageously, this selective precipitation step, for example flocculation, will make it possible to eliminate the molecules which are in suspension or in the form of a colloidal dispersion, and will thus make it possible to lower the quantity of dry matter, conventionally called dry extract, of the extract, while retaining the cardenolides which are soluble in the extraction solvent. Thus, the process according to the invention leads to an extract which is even more concentrated in cardenolides at the end of step c). Indeed, step c) will make it possible to increase the proportion represented by the cardenolides in the dry extract of the extract produced.

[0065] Step c) preferably consists of introducing into the mixture obtained in step a) or, in particular if step b) is carried out, into the liquid part L1 obtained in step b) at least one agent leading to the flocculation of solid materials, and is preferably followed by a step of eliminating the solid materials resulting from said flocculation.

[0066] Several approaches can be used to selectively remove a portion of the undesirable molecules from the liquid phase and enrich the cardenolide content of the non-volatile matter transferred from the biomass to the solvent. The undesirable compounds can be removed by an acid treatment by adjusting the pH of the medium to a value below 7, preferably below 4, using acids such as sulfuric acid, hydrochloric acid, phosphoric acid; a basic treatment, by adjusting the pH to a value below 10, using for example calcium hydroxide, potassium hydroxide, barium hydroxide, sodium hydroxide, especially in the presence of calcium chloride; and / or by using flocculating agents such as aluminum sulfate, iron sulfate, aluminum chloride, Per(III) chloride; or by using resins.

[0067] Step c) can thus be carried out by introducing into the mixture obtained in step a) or into the liquid part L1 obtained in step b), at least one agent leading to the flocculation of solid materials, the selective precipitation being preferentially carried out by flocculation of solid materials, for example by treatment with flocculation agents, by acid treatment or by basic treatment.

[0068] To do this, it is possible to use different agents, in particular: agents leading to the formation of an inorganic precipitate which will carry with it the suspended solids; as examples of such agents, we can cite the lime / phosphoric acid pair, and the barium hydroxide / sulfuric acid pair; flocculation agents by complexation; these agents lead to the formation of a complex with certain molecules in suspension which will then precipitate; as examples of such agents, we can cite lime; flocculation agents by modification of the pH; the modification of the pH obtained will accentuate the insoluble character of certain molecules; as examples of such agents, we can cite sulfuric acid.

[0069] In the context of the invention, the use of a lime / phosphoric acid couple, a barium hydroxide / sulfuric acid couple, lime alone, activated carbon and sulfuric acid are particularly preferred.

[0070] The lime / phosphoric acid and barium hydroxides / sulfuric acid couples are efficient flocculating systems, advantageously both in terms of the absence of degradation of cardenolides of interest, the reduction of the dry extract and its effectiveness regardless of the part of the plant used to constitute the starting plant raw material. The combined use of lime and phosphoric acid in particular leads to the formation of an inorganic precipitate (calcium phosphate) which carries with it the molecules which are still in suspension or in colloidal form, in the mixture obtained in step a) or the liquid part L1 obtained in step b).

[0071] Step c) can be carried out with stirring, in particular with mechanical stirring in the range from 15 to 1500 rpm.

[0072] The time taken to complete step c) to obtain satisfactory flocculation is in particular from 1 second to 30 minutes, once the selected agent(s) have been introduced.

[0073] Step d) then advantageously consists of carrying out one or more separations of the result of step c) to recover its liquid part L2.

[0074] The separation(s) carried out in step d) may be the same as those carried out in step b), such as filtration, centrifugation or decantation. It is particularly preferred to use centrifugation at this stage.

[0075] The separation of the solids resulting from flocculation and the remaining liquid part is conventionally carried out by any suitable separation device. In particular, the flocculated solids / liquid combination can be subjected to a filtration operation. It is also possible to carry out the separation of flocculated solids / liquid using a centrifuge, or by simple decantation. At the end of such a centrifugation or decantation step, the supernatant is recovered.

[0076] Thus, preferably, at the end of separation step d), the purified liquid part L2 is recovered and the flocculated solid matter eliminated.

[0077] Advantageously, step e) finally consists of carrying out a concentration of the liquid part L2 to recover an extract concentrated in cardenolides, for example by evaporation or filtration. The purified liquid part L2 can thus also be concentrated into dry extract, by at least partial evaporation of the extraction solvent or any other liquid present in the extract. For this, any suitable device, such as a reduced pressure evaporator, can be used. The extract can be concentrated by removing part of the solvent, for example by evaporation, if the solvent is water; by completely removing the solvent and replacing it with a certain quantity of water, if it is an organic solvent.

[0078] It is also possible to concentrate the liquid part L2 to recover an extract concentrated in cardenolides by federation, in particular membrane filtration, for example by nanofiltration or ultrafiltration.

[0079] It is also possible that the purified liquid part L2 is then completely dried or freeze-dried. In this case, the cardenolide-concentrated extract produced is the dry extract or the freeze-dried extract resulting from this drying or freeze-drying step.

[0080] These treatments will allow the extract to be further concentrated in cardenolides, while avoiding reaching excessively high viscosities.

[0081] The invention also relates to an extract obtained or capable of being obtained by the process as defined above.

[0082] Formulations

[0083] The invention also relates to a formulation comprising at least one extract as defined above and at least one additive, in particular chosen from the group consisting of: coating agents, binders, colorants, pigments, surfactants, dispersing agents, wetting agents, anti-foaming agents, rheological agents, anti-freeze agents, plasticizers, agents for promoting seed flow (such as waxes, graphite or oils), fillers, adjuvants, biocides, and active ingredients. The formulation of the invention may comprise one or more additives.

[0084] Preferably, the formulation comprises from 10 to 99% by weight of said extract, relative to the total weight of the formulation. The formulation may comprise from 30 to 99% by weight of said extract, relative to the total weight of the formulation, in particular when the extract is formulated with an active ingredient. The formulation may comprise from 90 to 99% by weight of said extract, relative to the total weight of the formulation, in particular when the extract is formulated with a dispersing agent and / or a wetting agent.

[0085] These additives, also sometimes called film-forming agents, are used to apply a microporous film to the surface of the seed. They improve seed coverage and the uniformity of coverage by the active ingredient. They can also facilitate seed flow in the seed drill.

[0086] Coatings can modify the shape, size and / or weight of the seed, thereby improving sowing accuracy. A single-layer coating can be carried out or the coating steps can be repeated to obtain a multi-layer coating. The formulation can also comprise one or more binders, in particular selected from the group consisting of gelatin, polyvinyl acetates, copolymers of polyvinyl acetate monomer with another monomer, polyvinyl alcohols, copolymers of polyvinyl alcohol monomer with another monomer, styrene-butadiene, styrene-carboxylated. butadiene, celluloses selected from the group consisting of ethylcelluloses and methylcelluloses, hydroxypropylcelluloses, hydroxymethylcelluloses and hydroxymethylpropylcelluloses, polyvinylpyrrolidones, dextrins, polysaccharides, fats, calcium oils, proteins, gum arabic, shellac, calcium oils, proteins, gum arabic, shellac,vinylidene chloride, vinylidene, chloride copolymers, vinylidene chlorides copolymers of acrylic monomers with another monomer, underivatized starch, derivatized starch, cationic starch, polyvinyl acrylates, zeins, carboxymethylcellulose, chitosan, polyethylene oxide, acrylimide polymers and copolymers of acrylimide monomer with another monomer, polyhydroxyethyl acrylate, methylacrylimide monomers, ethylcellulose, polychloroprene, guar, derivatized guar, polyacrylamide, poly(methacrylic acid), poly(acrylic acid), polyacrylate, poly(ethylene glycol), phosphonate-terminated polymers, polyethylene oxide, poly(vinyl alcohol), polyglycerol, polytetrahydrofuran, polyamides, cationic hydroxypropyl guar, hydroxypropyl guar, cationic carboxymethyl guar, guar carboxymethylhydroxypropyl, starch, derived starch, cationic starch, corn starch, wheat starch, rice starch, potato starch, tapioca, waxy maize,sorghum, waxy sarghum, sago, dextrin, chitin, chitosan, alginate compositions, xanthan gum, carrageenan gum, karaya gum, gum arabic, pectin, cellulose, hydroxycellulose, hydroxyalkylcellulose, hydroxyethylcellulose, carboxymethylhydroxyethylcellulose, hydroxypropylcellulose, a derivative of any of the foregoing and a combination of any of the foregoing.,

[0087] The binders may be, for example, acrylic polymers composed of one or more of the following monomers: acrylic acid, methyl acrylate, ethyl acrylate, butyl acrylate, hydroxyethyl acrylate, hydroxybutyl acrylate, carboxyethyl acrylate, methacrylic acid, methyl methacrylate, hydroxybutyl methacrylate, lauryl methacrylate, and stearyl methacrylate; with none and / or one or more of the following monomers: acrylamide, N-methylacrylamide, N,N-dimethylacrylamide, N-octylacrylamide, maleic anhydride, maleic acid, monoethyl maleate, diethyl maleate, monooctyl maleate, dioctyl maleate; and their corresponding sodium, potassium, ammonium, isopropylamine, triethylamine, monoethanolamine, and / or triethanolamine salts.Other suitable binders from this list include: copolymers of methyl vinyl ether with maleic anhydride or monoalkyl esters of maleic anhydride (e.g., ISP's Agrimer® VEMA product range); polyvinylpyrrolidones; copolymers of vinylpyrrolidone with vinyl acetate (e.g., ISP's Agrimer VA product range); copolymers of vinylpyrrolidone with vinylalkyls (e.g., ISP's Agrimer® AL product range); polyvinyl acetate; ethylene / vinyl acetate copolymers (e.g., Uniqema's Atlox® SemKote E product range); acrylic copolymers of vinyl acetate (e.g., Uniqema's Atlox® Semkote V product range); AB block copolymers of ethylene oxide and propylene oxide; ABA triblock copolymers of EO-PO-EO (e.g. BASF's Pluronic® range); polyvinyl alcohol, styrene acrylic polymers and vinyl acetate-versatate polymers.

[0088] Binders may also be a latex polymer, i.e., a dispersion in an aqueous carrier of polymer particles having a particle size of about 0.05-0.20 microns and a weight-average molecular weight greater than 500,000 g / mol. Latex polymers may be formed by conventional emulsion polymerization.

[0089] Binders may also be selected from biodegradable binders. Biodegradable binders may be water-soluble or water-insoluble and are generally degraded over a period of several weeks when placed in an application environment. Examples of biodegradable polymers include biodegradable polyesters; starch-polyester alloys; starch; starch-PCL blends; polylactic acid (PLA) starch blends; polylactic acid; poly(lactic acid-glycolic acid) copolymers; PCL; polygalactomannans, such as derivatized or underived guars; cellulose esters; cellulose acetate butyrate; starch esters; aliphatic starch-polyester ester blends; modified corn starch; polycaprolactone; poly(n-amyl methacrylate); ethylcellulose; wood rosin; polyanhydrides; polyvinyl alcohol (PVOH); polyhydroxybutyrate valerate (PHBV); biodegradable aliphatic polyesters; and polyhydroxybutyrate (PHB).

[0090] Guars may be guars or guar derivatives. Guar derivatives include, but are not limited to, carboxymethyl guar, hydroxyethyl guar, hydroxypropyl guar, carboxymethylhydroxypropyl guar, cationic guar, cationic hydroxypropyl guar, carboxymethyl guar, hydrophobically modified guar, hydrophobically modified carboxymethyl guar, hydrophobically modified hydroxyethyl guar, hydrophobically modified hydroxyethyl guar, hydrophobically modified cationically modified hydroxypropyl guar, hydrophobically modified carboxymethylhydroxypropyl guar, hydrophobically modified cationic guar, or any combination thereof. Starch or amylum is a carbohydrate consisting of a large number of glucose units linked by glycosidic bonds.

[0091] Binders can include vegetable oils such as linseed oil, soybean oil, rapeseed oil, and castor oil. These oils can be used as is or frozen and / or refined. These oils can contain triglycerides of various fatty acids, mostly unsaturated. These oils can optionally be ethoxylated.

[0092] Examples of pigments include: mica particles, titanium dioxide coated mica particles, titanium dioxide, phthalocyanine, chromium oxide, pigment red 112 (CAS No. 6535-46-2), pigment red 2 (CAS No. 6041-94-7), pigment red 48:2 (CAS No. 7023-61-2), pigment blue 15:3 (CAS No. 147-14-8), pigment green 36 (CAS No. 14302-13-7), pigment green 7 (CAS No. 1328-53-6), pigment yellow 74 (CAS No. 6358-31-2), pigment orange 5 (CAS No. 3468-63-1), pigment violet 23 (CAS No. 6358-30-1), pigment black 7 (CAS No. 97793-37-8), and white pigment 6 (CAS No. 98084-96-9).

[0093] In particular, ethoxylated tristyrylphenol phosphates (such as, for example, the products Soprophor® FL, Soprophor® FL / 60, Soprophor® FLK and Soprophor® 3D33), ethoxylated tristyrylphenol sulfates (such as, for example, the product Soprophor® 4D384), polycarboxylates (such as, for example, the products Geropon® SC / 213, Geropon® T / 36 and Geropon® TA / 72), methyl oleyl laurates (such as, for example, the product Geropon® T / 77), phenylsulfonates (such as, for example, the product Supragil® GN) and acrylic copolymers can be used as dispersing agents.

[0094] In particular, ethopropoxylated fatty alcohols (such as the product Antarox® FM / 33), ethoxylated fatty alcohols (such as the product Rhodasurf® CET 5, or the product Rhodasurf® 860 / P), sulfosuccinates (such as the products Geropon® DOS / PG and Geropon® SDS), alkylnaphthalene sulfonates (such as the product Supragil® WP) can be used as wetting agents (illustrative and non-limiting list).

[0095] Active ingredients may be, for example, one or more of the following: plant nutrients, growth-promoting agents and plant protection products.

[0096] According to the invention, the term "plant nutrient" includes any nutrient such as a micronutrient or a macronutrient. The term "nutrient" as used herein may refer to an additive or substance used by plants, grasses, shrubs for the growth of plants, grasses, and shrubs, respectively. Macronutrients may be used in larger amounts by plants, grasses, etc. in proportionately larger amounts compared to micronutrients. Nutrients include, but are not limited to, manganese, boron, copper, iron, chlorine, molybdenum and zinc, potassium, nitrogen, calcium, magnesium, phosphorus, and sulfur, among others. The formulations of the present invention may include various combinations and relative amounts of individual macronutrients.

[0097] According to the invention, the term “growth-promoting agents” includes biological additives, such as bacteria- or fungal-type inoculants, as well as plant biostimulants. Plant biostimulants are generally non-fertilizer components that affect plant growth and / or metabolism when applied foliarly, to the seed, or when added to the soil. Plant biostimulants generally fall into one of three categories: hormone-containing products, amino acid-containing products, and humic acid-containing products. Plant biostimulants are used to treat crops in a commercial setting due to their ability, for example, to increase growth rates, decrease the growth of harmful plants, increase stress tolerance, increase the rate of photosynthesis, and increase disease tolerance.Growth-promoting agents can also be microorganisms, for example. Microorganisms include, but are not limited to, Gram-positive bacteria, Gram-negative bacteria, including Rhizobia and Bacillus bacteria, and fungi, including Trichoderma.

[0098] Growth-stimulating agents may also be polysaccharides such as guars and cationic guars, as described in international applications WO2016 / 101862 and WO2021 / 074369.

[0099] According to the invention, the term "plant protection product" includes any pesticide such as, in particular, herbicides, fungicides or insecticides, preferably fungicides or insecticides. The pharmaceutical product may be a chemical substance, a biological agent (such as a macro-organism, a microorganism), a semiochemical product (such as pheromones) or natural substances of mineral, plant or animal origin used against pests, in particular insects, phytopathogens, weeds, molluscs, birds, mammals, fish, nematodes (roundworms) and microbes that compete with humans for food, destroy property, spread disease or are a nuisance. Pharmaceutical products include biopesticides.Suitable examples of active ingredients of the invention include fungicidal agents, bactericidal agents, insecticidal agents, nematicidal agents, molluscicidal agents, acaricides or miticides, pesticides and biocides. Other possible active ingredients include disinfectants, microorganisms, rodent killers, weedkillers (herbicides), attractants, repellents (for birds), plant growth regulators (such as gibberellic acid, auxin or cytokinin), nutrients (such as magnesium sulfate, iron chelate), plant hormones, minerals, plant extracts, germination stimulants, pheromones, biological preparations, etc.

[0100] The active ingredients may also be chosen, for example, from the group consisting of: triazine herbicides, sulfonylurea herbicides, uracils, urea herbicides, acetanilide herbicides, organophosphonate herbicides, glyphosate salts, glyphosate esters, nitrilo-oxime fungicides, imidazole fungicides, carbamate fungicides, chlorinated aromatics, dichloroaniline fungicides, carbamate insecticides, organothiophosphate insecticides; perchlorinated organic insecticides, methoxychlor, acaricides, propynyl sulfite, triazapentadiene acaricides, chlorinated aromatic acaricides, tetradifan, dinitrophenol acaricides, binapacryl, molluscicides, bactericides, repellents or any mixture thereof.

[0101] Additives can be added to the extract to obtain the formulation in various ways, such as by introducing them into a stirred tank to obtain a homogeneous mixture.

[0102] Among the various possible formulations, we can cite in particular: formulations comprising at least the extract according to the invention and a growth-stimulating agent of polysaccharide type, in particular chosen from guars and cationic guars. formulations comprising at least the extract according to the invention and a micronutrient formulation. formulations comprising at least the extract according to the invention and a film-coating formulation optimized for seed coating. formulations comprising at least the extract according to the invention and a biopesticide formulation for pest control.

[0103] The formulation according to the invention comprising at least one extract as defined above can perfectly be a bird repellent composition or a seed treatment spray. Coated seeds

[0104] The present invention also relates to a seed coated with an extract as defined above or a formulation as defined above.

[0105] Generally speaking, the active ingredients suitable for the desired effect should be applied evenly over the entire surface of the seeds. In fact, if the surface of the grain is not entirely covered by the products, including in the furrow, the protection could be less effective or even ineffective. In the present case, however, to the extent that the extract or formulation of the invention has a repellent effect, this effect is observed even if the entire seed is not covered.

[0106] The extract or formulation of the invention may be applied to seeds in various ways conventional in the art of seed treatment, including, but not limited to, mixing in a container, such as a bottle, bag or cup, mechanical application, spraying and immersion, optionally followed by drying.

[0107] Examples of seed coating techniques and machines that can be used include fluidized bed techniques, the roller mill method, rotary seed treatments, drum coaters, side-feed bins, drum mixers, and spout beds.

[0108] Seeds can be pre-sized before coating. The seed treatment mixture can be applied to the seeds in a seed treatment device, such as the Hege Seed Treatment Device, which rotates as the formulation is added to the seeds.

[0109] Mixing is preferably continued until the seed treatment formulation is evenly distributed over the seed (i.e., uniform coatings on all seeds to be treated and a uniform coating on each individual seed).

[0110] The seed treatment mixture may be applied to the seeds in a batch treatment process or in a continuous treatment process. In a representative batch treatment process, the seeds to be treated are introduced into a batch treatment tank and the treatment mixture is then added and mixed with the seeds. Alternatively, a continuous treatment process may be used to apply the seed treatment mixture to the seeds in which a stream of seeds is introduced into a receptacle containing the seed treatment suspension and, after contact with the formulation, recovered from the receptacle for drying.

[0111] Appropriate coating techniques can be used to coat seeds or seed agglomeration.

[0112] After the seed treatment mixture has been applied (whether in a batch or continuous process), the seeds are allowed to dry for a period of time. For example, the seeds may be turned in a bowl for a period of time, e.g., at least 15 seconds, to allow for drying. Different periods of time may be required to account for variability in drying conditions due to weather conditions or different seed sizes. In addition, heat may be provided, if desired, to reduce drying times, e.g., in the form of a heated air stream. After drying, the coated seeds may undergo a separation or size classification process.

[0113] Seeds may be separated before coating by mechanical means such as a sieve. The separated seeds may then be fed into a coating machine with a seed tank. The seeds in the mixing bowl may be combined with one or more coatings and bonded with a binder or adhesive.

[0114] The seeds of the plants according to the present invention may be agricultural or horticultural plants, of the monocotyledonous or dicotyledonous type, and may be planted for the production of an agricultural or horticultural product. The plant may be, for example, a cereal plant, in particular grasses.

[0115] The extracts and formulations of the invention can be applied to virtually any variety of plants and fruits. The plants can be selected from, but are not limited to, the following list:

[0116] - Food crops: such as cereals including maize / corn (Zea mays), sorghum (Sorghum spp.), millet (Panicum miliaceum, P. sumatrense), rice (Oryza sativa indica, Oryza sativa japonica), wheat (Triticum sativa), barley (Hordeum vulgare), rye (Secale cereale), triticale (Triticum X Secale), sunflower (Helianthus annuus) and oats (Avena fatua).

[0117] - Seeds can also be of any leguminous plant species, including, but not limited to, beans and peas. Beans include guar, carob, fenugreek, soybeans, garden beans, cowpeas, mung beans, lima beans, kidney beans, lentils, chickpeas, peas, broad beans, fava beans, kidney beans, lentils, and dry beans.Legumes include, but are not limited to, plants of the genus Arachis, e.g., peanuts, Vicia, e.g., crown vetch, hairy vetch, azuki bean, mung bean and chickpea, plants of the genus Lupinus, e.g., lupin, trifolium, plants of the genus Phaseolus, e.g., common bean and lima bean, plants of the genus Pisum, e.g., faba bean, plants of the genus Melilotus, e.g., clover, plants of the genus Medicago, e.g., lucerne, plants of the genus Lotus, e.g., clover, lentil, e.g., lentil and false indigo. Forage and turf include, but are not limited to, alfalfa, tall fescue, perennial ryegrass, creeping bentgrass, lucerne, clover, clover, stylosanthes, Lotononis bainesii, sainfoin, and white bentgrass. Other grass species include barley, wheat, oats, rye, orchardgrass, guinea fowl, sorghum, or turf.

[0118] - Leafy vegetables: such as brassica plants such as cabbage, broccoli, bok choy, arugula; green salads such as spinach, watercress, basil and lettuce;

[0119] - Root vegetables: such as radishes, carrots, beets and celeriac.

[0120] - Fruiting and flowering vegetables: such as sweetcorn, artichokes, cucurbits, e.g. squash, cucumbers, melons, watermelons, squash such as zucchini, pumpkins; solononaceous vegetables / fruits, e.g. tomatoes, aubergines and peppers; - Podded vegetables: such as peanuts, peas, beans, lentils, chickpeas and okra;

[0121] - Bulb and stem vegetables: such as celery, allium crops, e.g. garlic, onions and leeks;

[0122] Coating can also be carried out on barley, rye, corn, rapeseed, soybean or sunflower seeds, but this method can also be applied to cucurbit, solanaceous, brassica or asteraceae vegetable species.

[0123] The dosage of extracts or formulations on seeds can vary depending on various parameters such as the nature of the extract and the type of seeds. For example, approximately 0.1 to 3 kg, preferably 0.1 to 2 kg, of extracts or formulations can be used for 1 quintal (q) of seeds.

[0124] The coated seeds according to the invention may in particular be coated with an extract or a formulation according to the invention and a composition comprising at least one film-coating agent. These two compositions may be mixed beforehand before application to the seeds. These two compositions may be applied successively or at the same time to the seeds.

[0125] Use of an extract or formulation

[0126] The invention also relates to the use of an extract as defined above or of a formulation as defined above, for treating seeds.

[0127] The invention also relates to the use of an extract as defined above or of a formulation as defined above, as a bird repellent.

[0128] The solution also has an effect against rodents and other domestic pests, thus providing double protection for treated seeds. The use of this solution for its repellent effect against birds and rodents can be extended to other areas of activity.

[0129] The invention also relates to the use of the extracts and formulations as a phytosanitary product, or the use of the extracts and formulations for the preparation of a phytosanitary product.

[0130] The following examples illustrate the invention without, however, limiting its scope.

[0131] The presence of the cardenolides of interest was confirmed:

[0132] - By UV spectrometry, using the equipment and protocol below: Agilent 1290,

[0133] Column: Agilent model - ZORBAX Eclips XDB Cl 8, Dimensions: 150 mm x 3.0 mm x 5 im,

[0134] Mobile phase:

[0135] A: Water + 0.1% v / v Formic Acid and B: Acetonitrile + 0.1% v / v Formic Acid

[0136] The gradient presented in Table 1 below:

[0137] Table 1:

[0138] Flow rate: 0.3 mL / min,

[0139] 5 pl injection,

[0140] UV detection from 190 to 600 nm (2 nm step).

[0141] - By liquid phase mass spectrometry (high resolution LC-MS) using in particular the equipment and protocol below:

[0142] Agilent QTOF,

[0143] Positive Electrospray Source,

[0144] Fragmenter voltage: 50 V,

[0145] Gas temperature: 325°C,

[0146] Drying gas flow rate: 8 L / min

[0147] Nebulizer pressure: 45 psig,

[0148] Absorption gas temperature: 300°C,

[0149] Absorption gas flow rate: 11 L / min,

[0150] Capillary voltage: 3000 V,

[0151] Nozzle voltage: 0 V,

[0152] CID: Nitrogen, 4 V ramp and 10 V offset.

[0153] The quantity of cardenolides of interest was determined by mass spectrometry. For this, the extract was diluted at a rate of 25 to 30 mg of extract for 20 g of water. If necessary, in case of formation of a precipitate, during the preliminary steps of storage of the extract, in particular in frozen form, the extract was passed through an ultrasonic bath, to homogenize the sample.

[0154] In high-resolution LC-MS, the relevant data used for the detection of the [M+H] ion + , adducts and majority fragments for each of the four cardenolides of interest are presented in Table 2 below:

[0155] Table 2:

[0156] * TR = Retention Time (time after which a compound is eluted and detected)

[0157] Example 1: Preparation of the extract, method 1

[0158] 4.14 kg of dried biomass of crushed Asclepias curassavica leaves and stems were introduced into a tank containing 41.4 kg of water at 30 °C. The medium was kept stirring at 30 °C for 3 hours.

[0159] After maceration, the liquid phase was separated from the biomass by a centrifugal decanter.

[0160] 36.1 kg of diluted juice with a dry extract of 2.44% was separated from the biomass. The biomass had a moisture content of 69.77%.

[0161] The pH of the diluted extract was 5.9. Approximately 0.27 kg of 75% by mass phosphoric acid was added to obtain a pH of 2.8. The solution was stirred and kept at 30 °C for 30 minutes and the formation of flocs in the mixture was observed. The flocculated products were removed by centrifugation and 35.6 kg of clear juice with a dry extract of 2.65% was obtained. The pH of the juice was 2.7.

[0162] 0.7 kg of 30% by mass lime milk was then introduced and led to the formation of hydroxyapatite and other calcium phosphate species almost instantaneously. The precipitate was removed by centrifugation. 33.7 kg of clarified juice with a dry extract of 2.06% was obtained.

[0163] 80 g of 3% by mass lime milk was added to the clarified juice to adjust the pH to 7.1. Hydroxyapatite was precipitated a second time. The precipitate was removed by centrifugation. 32.9 kg of juice with a dry extract of 1.97% was obtained. After 12 hours of storage at 30 °C, the clear juice became turbid again with a drop in pH to 4.7. The juice was centrifuged again. 29.34 kg of clarified juice with a dry extract of 1.88% was recovered after centrifugation.

[0164] The diluted juice was then concentrated by distilling the water in a falling-flow evaporator.

[0165] 76.2 kg of concentrated juice with a dry extract of 64.2% was obtained. The cardenolide content was 1.13% by weight of cardenolides, relative to the total weight of said extract, measured by liquid chromatography coupled with mass spectrometry. Uscharidin, calotoxin, asclepin, calotropagenin, calactin, uscharin, 12p-hydroxycoroglaucigenin, 12p-hydroxycalactropin, 15 P-hydroxy calactin and gofruside were detected. To measure the viscosity, 300 g at 30 °C of extract were weighed into a 600 mL beaker (height 12.4 cm and diameter 9 cm). A Brookfield type RVDV viscometer was used at 20 rpm with a needle to be in the correct range for a reliable measurement. The viscosity of the extract was measured at 358 mPas.

[0166] Example 2: Preparation of the extract, method 2

[0167] 42.2 g of dried biomass of crushed Asclepias curassavica leaves and stems were introduced into a tank containing 420 g of water at 30 °C. The medium was kept stirring at 30 °C for 3 hours.

[0168] After 3 hours, 7.8 g of 85% phosphoric acid by mass was added to the mixture to lower the pH from 5.63 to 2.03. The solution was then stirred for 5 minutes; then 21 g of 30% milk of lime was added to adjust the pH to 7.32.

[0169] After pH adjustment, the liquid phase was separated from the solid phase by centrifugation. 253 g of juice with a dry extract of 1.71% by weight was recovered.

[0170] The diluted juice was then concentrated by water distillation in a rotary evaporator. 6.8 g of concentrated juice with a dry extract of 64% was obtained. The cardenolide content of the plant extract was 1.07% by mass measured by liquid chromatography coupled with mass spectrometry. Uscharidin, calotoxin, asclepin, calotropagenin, calactin, uscharin, 12p-hydroxycoroglaucigenin, 12p-hydroxycalactropin, 15p~ hydroxy calactin and gofruside were detected.

[0171] To measure viscosity, 300 g of extract at 30 °C was weighed into a 600 mL beaker (height 12.4 cm and diameter 9 cm). A Brookfield RVDV type viscometer was used at 20 rpm with a needle to be in the correct range for a reliable measurement. The viscosity of the extract was measured at 370 mPas.

[0172] Example 3: Preparation of the extract, method 3

[0173] 1186 g of crushed Asclepias curassavica leaves were introduced into a tank containing 11.85 kg of water at 30 °C. The medium was kept stirring at 30 °C for 3 hours.

[0174] After maceration, the liquid phase was separated from the biomass by centrifugation. 8.15 kg of diluted juice with a dry extract of 2.84% was separated from the biomass.

[0175] The diluted juice was then concentrated by vacuum distillation of water. 450 g of concentrated juice with a dry extract of 50% was obtained. The cardenolide content of the plant extract was 1.6% by weight measured by liquid chromatography coupled with mass spectrometry. Uscharidin, calotoxin, asclepin, calotropagenin, calactin, uscharin, 12p-hydroxycoroglaucigenin, 12p-hydroxycalactropin, 15 P-hydroxy calactin and gofruside were detected. To measure the viscosity, 300 g of extract at 30 °C was weighed into a 600 mL beaker (height 12.4 cm and diameter 9 cm). A Brookfield type RVDV viscometer was used at 20 rpm with a needle to be in the correct range to have a reliable measurement. The viscosity of the extract was measured at 370 mPas.

[0176] Example 4: Preparation of a diluted, purified extract

[0177] 4 g of crushed Asclepias curassavica leaves were introduced into a tank containing 40 g of water at 30 °C. The medium was kept stirring at 50 °C for 3 hours.

[0178] After maceration, the liquid phase was separated from the biomass by centrifugation. 30 g of diluted juice with a dry extract of 3.73% was separated from the biomass.

[0179] The pH of the diluted extract was 6.1. Approximately 0.8 g of FcCL in water dispersion at 0.6% by mass was added. The solution was stirred and kept at 30 °C for 10 minutes and the formation of flocs in the mixture was observed. The flocculated products were removed by centrifugation and 27 g of clear juice with a dry extract of 1.96% was obtained.

[0180] Example 5: Preparation of extracts

[0181] Table 3 below presents the characteristics of two extracts, one made without “purification” (Sample A), the other including a purification step (Sample B).

[0182] To prepare sample A, 100 g of crushed dried biomass was macerated in 1 liter of water for 3 hours. The liquid phase was separated from the solid phase by centrifugation. 700 g of juice with a dry extract of 2.4% was recovered. The juice was then concentrated to obtain 33 g of juice with a solid content of 50%.

[0183] To prepare sample B, 100 g of dried ground biomass was macerated in 1 liter of water for 3 hours. 85% phosphoric acid was added to lower the pH of the mixture to 2.7. The mixture was stirred for 10 minutes. Then, milk of lime was added to raise the pH from 2.7 to 7.2. The mixture was stirred for 10 minutes. Then the liquid phase was separated from the solid phase by centrifugation. 720 g of juice with a solids content of 2.0% was recovered. The juice was then concentrated to obtain 22.5 g of juice with a solids content of 64%. Table 3

[0184] It is thus observed that the purification carried out makes it possible to increase the cardenolide content in the dry matter while having a lower viscosity and therefore a product that is more easily handled for the same dry matter content. In addition, the dose to be applied to seeds is lower, and therefore easier to implement on an industrial scale.

[0185] Example 6: Formulation of the extract

[0186] To protect the extract from microbial growth, 76.2 g of kathon™ CG (CIT / MIT) were added to the extract obtained in Example 1. To ensure stability of the formulation over time, 0.57 kg of an ethoxylated fatty alcohol such as Rhodasurf® 860 / P and 1.5 kg of etho (30 EO)-propoxylated (20 PO) tristyrylphenol such as Soprophor® TSP / 461 were added. The pH was readjusted to 7 using potassium hydroxide.

[0187] The cardenolide content of the resulting formulation was 1.08% by weight, measured by liquid chromatography coupled with mass spectrometry. The viscosity of the formulation is 1000 mPas.

[0188] To measure viscosity, 300 g of extract at 25 °C was weighed into a 600 mL beaker (height 12.4 cm and diameter 9 cm). A Brookfield type RVDV viscometer was used at 20 rpm with a needle to ensure a reliable measurement. Example 7: Seed treatment for aviary trials

[0189] Seed treatment slurries containing a seed treatment fungicide (Redigo® M from Bayer), a film-coating agent, water and an extract (obtained according to method 1 presented in Example 1 or according to method 3 presented in Example 3) are prepared by mixing all the compounds by mechanical agitation. A slurry without the addition of the extract is prepared; this is the “Control” treatment. A slurry is also prepared with a commercial bird repellent (Korit 420 FS) to serve as a point of comparison; this is the “Reference” treatment. The exact compositions of the different seed treatment slurries are presented in Table 4. Corn seeds (Zea mays, commercially sourced, PMG 275) are treated in a multi-layer manner, by applying several successive layers with an intermediate drying time (2 hours) using a SATEC bowl coater.Seed treatment sprays are sprayed onto the seeds kept rotating in the bowl for a few seconds.

[0190] Table 4

[0191] Example 8: Aviary test results

[0192] Tests were carried out on birds in aviaries according to the protocol described in WO 2020 / 109529 examples 5 to 7 on the treatments presented in Example 7.

[0193] The experiment is carried out on crows and rooks in aviaries (7 individuals per aviary, Crow and Rook). The test is carried out over three days for each treatment tested (TRI, T2 and T3): On day 1, a balanced diet is offered to the birds. This diet is composed of 20 g of kibble and corn treated with the Control treatment (TCI) at will.

[0194] On day 2, 50 g of corn treated with the Control treatment (TCI) are offered.

[0195] On day 3, 20 g of corn treated with the bird repellent formulation (TRI, T2 or T3) are offered. On days 2 and 3, the refused feed is weighed at the end of the day to calculate the average consumption per animal.

[0196] The tests were conducted in parallel on two aviaries, which thus constitute two repetitions of the experiment. The results of the tests for the different treatments tested are presented in Table 5. For each treatment, the consumption difference was calculated as follows:

[0197] Consumption gap

[0198] Quantity but control consumed — Quantity but treated consumed

[0199] Quantity but control consumed

[0200] Table 5

[0201] As detailed in Table 5, the use of the extract formulation (T2 or T3) in the porridges leads to a reduction in the quantity of seeds consumed by the birds, this reduction is on average of the order of 22-23%.

[0202] Example 9: Seed treatment for field trials

[0203] Seed treatment slurries containing a seed treatment fungicide (Redigo® M from Bayer), a film-coating agent, water, and the extract of Example 3 at two different doses are prepared by mixing all the compounds by mechanical agitation. A slurry without the addition of the extract is prepared; this is the “Control” treatment. A slurry is also prepared with a commercial bird repellent (Korit 420 FS) to serve as a comparison point; this is the “Reference” treatment. The exact compositions of the different seed treatment slurries are presented in Table 6. Corn seeds (Zea mays, commercially sourced, PMG 326) are treated in a multi-layer manner, by applying several successive layers with an intermediate drying time (2 hours) using a SATEC bowl coater. The seed treatment slurries are sprayed onto the seeds, which are kept rotating in the bowl for a few seconds.

[0204] Table 6

[0205] Example 10: Field test results

[0206] Two field trials were conducted in Italy in the Turin region (Italy) to evaluate the bird repellent performance of the different treatments presented in Table 6. The field trials were carried out on microplots with 4 replicates per treatment. The size of the microplots was between 30 and 40 m 2 . Field trials were conducted following local agricultural practices (application of pesticides, fertilizers and irrigation). The inputs applied were exactly the same for the different microplots of the same trial. In terms of scoring, the number of germinated seeds on the two central rows was determined for each of the microplots at the BBCH 13 development stage of the maize plants, corresponding to the stage of 3 unfolded leaves.

[0207] Tables 7 and 8 present the results of the field trials. For each treatment, the total number of germinated seeds is indicated. This value corresponds to the average of the four microplots for each treatment. A statistical analysis of the results (Student-Newman-Keuls mean comparison test at 5%) was performed to determine whether the means are statistically different. The result of the statistical analysis is shown in the following tables using the letters in parentheses representing the statistical groups. Means that do not share a letter are statistically different.

[0208] Table 7: Castagnito test results - Number of germinated seeds

[0209] Treatments T4 and T5 (statistical group (a)) resulted in a significant increase in the number of germinated seeds compared to the control treatment (TC2, statistical group (b)) without bird repellent. The relative increase in the number of germinated seeds was approximately 18% for both treatments (T4 and T5). Finally, for both treatments the number of germinated seeds was comparable to the value obtained for the treatment with the reference bird repellent product (TR2, Korit 420FS, statistical group (a)).

[0210] Table 8: Lombriasco test results - Number of germinated seeds Treatments T4 and T5 (statistical group (a)) result in a significant increase in the number of germinated seeds compared to the control treatment (TC2, statistical group (b)) without bird repellent. The relative increase in the number of germinated seeds is between 15 and 20% depending on the treatment considered (T4 or T5). Finally, for both treatments the number of germinated seeds is comparable to the value obtained for the treatment with the reference bird repellent product (TR2, Korit 420 FS, statistical group (a)).

Claims

DEMANDS 1 - Extract including at least: - 0.5 to 4% by weight of cardenolides, relative to the total weight of said extract, - 10 to 85% by weight of dry matter, relative to the total weight of said extract, - 1 to 8% by weight of cardenolides, relative to the total weight of said dry matter, said extract having a viscosity between 1 and 10000 mPa.s. 2 - Extract according to the preceding claim, characterized in that it corresponds to an extract of at least one plant containing cardenolides. 3 - Extract according to any one of the preceding claims, characterized in that it comprises from 1 to 4% by weight of cardenolides, relative to the total weight of said extract. 4 - Extract according to any one of the preceding claims, characterized in that it comprises from 1.25 to 4.75% by weight of cardenolides, relative to the total weight of its dry matter. 5 - Extract according to any one of the preceding claims, characterized in that the cardenolides are selected from the group consisting of: calotoxin, uscharidine, asclepin, calactin, calotropagenin, coroglaucigenin, laxysmalogenin, voruscharin, gofruside, uzarigenin, 12p-hydroxy coroglaucigenin, 12p-hydroxy calotropin, 6'-O-feruloyl-desglucouzarine, 12p,14p-dihydroxy-3p,19-epoxy-3a-methoxy-5a-card-20(22)-enolide, laxysmalogenin, coroglaucigenin, 6-O-(E-4-hydroxycinnamoyl)-desglucouzarine, 6-O-sinapinoyl-desglucouzarine, calactin, 16α-acetoxycalactin, calotropin, 16α-acetoxycalotropin, 16α-hydroxyasclepin, 16α-acetoxyasclepin, uscharin, 16α-hydroxyuscharin, 19-nor-16α-acetoxy-10p-hydroxyasclepin, desglucouzarine, 6'-O-feruloyl-desglucouzarine, and calotropin 6 - A process for obtaining an extract of at least one plant containing cardenolides, comprising at least the following successive steps: a) bringing a raw material of at least one plant containing cardenolides into contact with a cardenolide extraction solvent S, b) optionally carrying out a separation of the result of the contact in step a) to recover its liquid part L1, c) carrying out a selective precipitation of the constituents other than cardenolides of the mixture obtained in step a) or of the liquid part L1 obtained in step b), d) carrying out one or more separations of the result of step c) to recover its liquid part L2, and e) carrying out a concentration of the liquid part L2 to recover a concentrated extract of cardenolides. 7 - Process according to claim 6, characterized in that the extraction solvent S is water or an aqueous solution. 8 - Process according to claim 6 or 7, characterized in that the separation in step b) is carried out by filtration, centrifugation or decantation. 9 - A process according to any one of claims 6 to 8, characterized in that the selective precipitation is carried out by flocculation of solid materials, for example by treatment with flocculating agents, by acid treatment or by basic treatment. 10 - A process according to any one of claims 6 to 9, characterized in that the selective precipitation is carried out using a flocculating agent such as aluminium sulfate, iron sulfate, aluminium chloride, iron(III) chloride; or alternatively using resins. 11 - A process according to any one of claims 6 to 10, characterized in that the separation in step d) is carried out by filtration, centrifugation or decantation. 12 - A process according to any one of claims 6 to 11, characterized in that the concentration at step e) is carried out by evaporation or filtration. 13 - Extract obtained or capable of being obtained by the process as defined in any one of claims 6 to 12. 14 - Formulation comprising at least one extract as defined according to any one of claims 1 to 5 or 13, and at least one additive. 15 - Formulation according to claim 14, characterized in that the additive is selected from the group consisting of: coatings, binders, colorants, pigments, surfactants, dispersing agents, wetting agents, antifoaming agents, rheological agents, antifreeze agents, plasticizers, seed flow promoters, fillers, adjuvants, biocides, and active ingredients. 16 - Formulation according to claim 14 or 15, characterized in that the formulation comprises from 10 to 99% by weight of said extract, relative to the total weight of the formulation. 17 - Formulation according to any one of claims 14 to 16, characterized in that the formulation comprises from 30 to 99% or from 90 to 99% by weight of said extract, relative to the total weight of the formulation. 18 - Formulation according to any one of claims 14 to 17, characterized in that the additive is an active ingredient, in particular selected from plant nutrients, growth-stimulating agents and plant protection products. 19 - Formulation according to claim 18, characterized in that the additive is a growth-stimulating agent consisting of a polysaccharide, in particular selected from guars and cationic guars. 20 - Seed coated with an extract as defined in any one of claims 1 to 5 or 13 or with a formulation as defined in any one of claims 14 to 19. 21 - Coated seed according to claim 20, characterized in that it is coated with an extract as defined in any one of claims 1 to 5 or 13 or with a formulation as defined in any one of claims 14 to 19 and a composition comprising at least one film-coating agent. 22 - Use of an extract as defined in any one of claims 1 to 5 or 13 or of a formulation as defined in any one of claims 14 to 19, for treating seeds. 23 - Use of an extract as defined in any one of claims 1 to 5 or 13 or of a formulation as defined in any one of claims 14 to 19, as a bird repellent. 24 - Use of an extract as defined in any one of claims 1 to 5 or 13 or of a formulation as defined in any one of claims 14 to 19, as a plant protection product or for the preparation of a plant protection product.

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