Process for transforming biowaste and by-products from the agri-food industry into biosolutions

Abstract

The present invention relates to the field of agriculture and more particularly to the manufacture of biosolutions intended to improve crop fertilization, crop protection, and animal health. The invention specifically concerns a process for transforming biowaste and by-products from the agri-food industry into biostimulants.

Description

Title of the invention: Process for transforming biowaste and by-products from the agri-food industry into biosolutions. Technical field

[0001] The invention relates to the manufacture of biosolutions for improving crop fertilization, crop protection, and animal health. More particularly, the invention relates to a process for transforming biowaste and by-products from the agri-food industry into biosolutions for agriculture and food, including biocontrol products, biostimulants, biofertilizers, agri-food preservatives, post-harvest preservatives or plant protection products, and probiotics. State of the art

[0002] Environmental issues are a major concern in our societies. It is therefore necessary to fundamentally rethink our economic and social models in order to combat climate change, the collapse of biodiversity, the depletion of natural resources, and the increase in environmental risks.

[0003] In this context, reducing the use of plant protection products and synthetic or fossil-based inputs is a strong public expectation and a necessity for preserving our health and biodiversity. As such, most governments, particularly in the West, are taking measures, including regulatory ones, to meet these expectations.In France, for example, the Ecophyto plan aims to reduce and / or improve the use of fertilizers and plant protection products, accelerate the withdrawal of the most concerning substances, strengthen the prevention of the population's exposure to pesticides and their impacts on the environment and biodiversity, revitalize soil biodiversity and symbioses between plants and microorganisms, a source of adaptation and resilience to biotic and abiotic stresses accelerated by climate change, promote the recognition and dissemination of biocontrol products and natural preparations, and also support farmers in this transition.

[0004] There is therefore a need for new products designed to improve crop fertilization, crop protection and animal health, to accelerate the replacement of synthetic molecules derived from petrochemicals and fossil minerals by renewable molecules from green chemistry, microorganisms of agronomic interest or natural substances with a lower impact on the Environmental quality and human health are also key. This allows us to more generally address the need to preserve our health and biodiversity, maintain our agricultural production capacity, and thus ensure the economic and environmental resilience of our farms. Finally, in the current context, there is also a need to develop circular economy sectors that will allow us to replace synthetic or fossil-based inputs with bio-based and low-carbon biosolutions. Summary of the invention

[0005] To meet these needs, the invention proposes to use materials, considered as bio-waste or by-products from the renewable agri-food industry, which are largely under-exploited to date, as raw materials for the manufacture of biosolutions, such as biocontrol products, biostimulation products, technological additives, biological silage preservatives, zootechnical additives, agronomic additives, biofertilizers, agri-food preservatives, post-harvest preservatives or plant protection products, as well as probiotics.

[0006] By using biowaste or by-products from the agri-food industry, a circular economy sector is created to valorize and synthesize high-value-added molecules present in this currently untapped raw material and subsequently transform them into biosolutions for agriculture, thus addressing the need to move away from synthetic molecules known for their impact on biodiversity and health. Furthermore, the use of biowaste or by-products from the agri-food industry makes it possible to implement a low-carbon, short-circuit process without the addition of petrochemical solvents and fermentable sugars.

[0007] In this context, the inventor has developed a new low-carbon process for transforming this unvalued raw material comprising several transformation steps.

[0008] Thus, the invention relates to a process for transforming biowaste and by-products from the agri-food industry into biosolutions intended to improve fertilization and / or crop protection, or animal health, said process comprising at least the following steps: a. Collection of biowaste b. Physical separation of the solid and liquid fractions of the collected biowaste, c. Recovery of the liquid fraction, and d. extraction of the molecules of interest, preferably present in said liquid fraction or the mixture comprising the liquid and solid fractions, and / or e. Fermentation comprising the addition of at least one exogenous microorganism of interest, preferably at least one mesophilic and / or homofermentative microorganism, f. Stabilization of the fermented product from the previous step, g. Optionally, filtration of the stabilized product.

[0009] The separation step, in particular a physical separation using specific devices, separates the different phases or components of the biowaste. Preferably, this separation step is a centrifugation-sedimentation step, carried out using a decanter-centrifuge. More preferably, it is a continuous flow centrifugation-sedimentation step at a flow rate of no more than 100 hectoliters / hour.

[0010] Next, the extraction step of the molecules of interest is carried out using green chemistry, that is to say, without the use of petrochemical solvents or the synthesis of hazardous substances. Advantageously, the extraction step comprises the following substeps: cold maceration with an alcoholic solvent derived from natural fermentation, followed by decoction, preferably at a temperature of no more than 80°C.

[0011] According to one embodiment, the extraction step may be replaced or followed by a fermentation step. This fermentation step uses the fermentable carbons contained in the liquid fraction to propagate said exogenous microorganisms, which themselves generate organic metabolites of interest. Thus, the present fermentation step does not involve the addition of exogenous sugars or dietary sugars (e.g., beet sugars, cane sugar, etc.) as fermentable carbons. Advantageously, this step is a fermentation step of simple and complex sugars from locally sourced biowaste by microorganisms under mesophilic conditions, enabling a so-called low-carbon process.

[0012] Preferably, the process according to the invention is thus low carbon, low energy consumption and low external resources other than bio-waste or by-products from the agri-food industry.

[0013] Preferably, to increase the yield, the fermentation is carried out at a temperature between 17 and 37 °C and / or a pH between 3 and 7, more preferably between 5 and 6.

[0014] The fermentation step advantageously allows the conversion of biowaste or agri-food by-products into new products known as biosolutions, whether a biocontrol product, a biostimulant, a biofertilizer, or a probiotic containing their own active ingredients. To achieve this, the inventor has advantageously Isolated and selected strains of exogenous microorganisms with fermentation capacities suited to the culture medium, i.e., the liquid fraction on the one hand, and the solid fraction on the other. According to a particular embodiment, the liquid fraction is capable of producing metabolites of interest depending on the final product of interest.

[0015] In the context of the invention, the exogenous microorganism of interest is preferably chosen from a lactic acid ferment, a bacterium of a genus chosen from Bacillus, Lactobacillus, Pseudomonas, Acetobacter, Nitrobacter, Nitrosomonas, Rhizobium, Agrobacterium, and Streptomyces, a fungus of the genus Trichoderma, a yeast of the genus Saccharomyces, or Aureobasidium, and their combinations.

[0016] According to a preferred design, the stabilization of the extracted and / or fermented product is carried out either by heat treatment or by cold treatment, such as microfiltration, UV filtration, or acid treatment. More preferably, the heat treatment is hot stabilization between 75 and 80°C, or the acid treatment is a dosage selected from acetic, peracetic, propionic, and lactic acids. Stabilization by microfiltration, UV filtration, or acid treatment makes it possible, in particular, to inactivate or eliminate microorganisms in the stabilized products. Such a stabilization step thus makes it possible to meet regulatory requirements for products containing no live microorganisms, for example, the specifications for Low-Risk Natural Preparations and Natural Substances for Biostimulant Use.

[0017] According to a preferred object of the invention, the raw material, i.e. bio-waste or by-products from the agri-food industry, is chosen from brewing by-products, cider by-products, wine by-products, olive oil by-products, distillery by-products, cocoa by-products, coffee by-products, starch factory by-products, sugar factory by-products, fruit juice press by-products, and dairy and cheese industry by-products, and combinations thereof.

[0018] According to an object, the brewing by-products are chosen from brewing hop dregs, fermentation yeast, fermentation hop purge or guard, malt dregs, brewery wastewater, distillery wastewater and their mixture.

[0019] According to an object, the wine by-products are chosen from vinasse, lees, grape seeds, and their mixture.

[0020] According to an object, the distillery by-products are selected from the residual distillation water, the fermentation yeast, the malt draff and their mixture.

[0021] According to an object, cider by-products are chosen from apple pomace, lees, and their mixture.

[0022] According to an object, the olive by-products are chosen from the olive mill wastewater, the pomace, and their mixture.

[0023] According to an object, cocoa by-products are selected from cocoa liquor, cocoa cake, and their mixture.

[0024] According to one object, the by-product of coffee is coffee grounds.

[0025] According to an object, the starch by-products are selected from the peels, the wastewater, and their mixture.

[0026] According to an object, the by-products of the sugar mills are chosen from molasses, vinasse, and their mixture.

[0027] According to an object, the by-products of fruit juices are chosen from pomace, citrus peels, and their mixture.

[0028] According to another preferred object of the invention, the final product, that is to say the transformed product obtained according to one of the embodiments of the process according to the present invention, is a biosolution product chosen from biocontrol products, biostimulation products, technological additives, biological silage preservatives, zootechnical additives, agronomic additives, biofertilizers, agri-food preservatives, post-harvest preservatives or plant protection products, and probiotics.

[0029] According to another particularly preferred aspect of the invention, the inventor has identified specific pairs or complexes (biowaste / microorganisms). The combination and complementarity of the selected microorganisms and raw material enhance its effectiveness and the manufacturing yield of the final biosolution product.

[0030] Thus, according to a particular embodiment of the invention, the biosolution is a biostimulation product or a biofertilizer and the exogenous microorganism of interest is a bacterium belonging to the genus chosen from Lactobacillus, Bacillus, Trichoderma, Saccharomyces, Acetobacter, and their combinations.

[0031] According to another particular embodiment of the invention, the biosolution is a biocontrol product and the microorganism is chosen from Bacillus, Trichoderma, Streptomyces, Pseudomonas, Aureobasidium, Saccharomyces, and their combinations.

[0032] According to another particular embodiment of the invention, the biosolution is a zootechnical additive or a probiotic and the exogenous microorganism of interest is chosen from Bacillus, Lactobacillus, Saccharomyces and their combinations.

[0033] According to another particular embodiment of the invention, the biosolution is an agri-food additive, or a silage preservative, or a food preservative and the exogenous microorganism of interest is chosen from Bacillus, Lactobacillus, Saccharomyces and their combinations.

[0034] Other features and advantages will become apparent from the detailed description of the invention, the examples and figures that follow. Brief description of the Figures

[0035] [Fig.1] represents the efficacy results of three biosolutions according to the invention against downy mildew of the vine. Detailed description of the invention

[0036] Definition

[0037] For the purposes of this invention, "biosolutions" refers to a range of products that enable farmers to produce. Biosolutions include biofertilizers, which allow crops to better absorb nutrients while preserving soil resources through the rhizosphere; biostimulants, which stimulate plant defenses against abiotic stresses; biocontrols, which mimic nature to protect plants against biotic stresses; and technological additives or complexes of microorganisms, which promote the transformation of organic matter from an unstable state to a more stable state, ensuring its optimal preservation or a quality that meets a standard for a defined use.

[0038] For the purposes of this invention, "biowaste" or "by-products from the agri-food industry" means organic waste necessarily originating from the agri-food industry, particularly from natural plant or animal resources that are also putrescible, fermentable, hydrolyzable, or oxidizable. Preferably, biowaste is not household waste suitable for composting, or organic or domestic effluents, or wastewater. More preferably, these are brewing by-products, cider by-products, wine by-products, olive oil by-products, distillery by-products, cocoa by-products, coffee by-products, starch factory by-products, sugar factory by-products, fruit juice press by-products, and dairy and cheese industry by-products.

[0039] For the purposes of this invention, a "complex" or "pair" refers to the combination and complementarity of specific and selected microorganisms with a specifically selected raw material in order to valorize the molecules of biowaste and agri-food by-products into various metabolites of agronomic interest through fermentation, hydrolysis, or oxidation, advantageously in a synergistic manner between the metabolites produced by the microorganisms during this transformation and the plant-based active ingredients contained in the same biowaste or by-product that served as the culture medium. Such a complex makes it possible to obtain a biosolution product of interest. For example, the combination of Bacillus and apple pomace can be used to obtain a biocontrol product.

[0040] Transformation process

[0041] The present invention therefore relates to a process for transforming biowaste and by-products from the agri-food industry into biosolutions intended to improve fertilization and / or crop protection, or animal health, said process comprising at least the following steps: a. Collection of biowaste b. Physical separation of the solid and liquid fractions of the collected biowaste, c. Recovery of the liquid fraction, and d. Extraction of the molecules of interest present in said liquid fraction, and / or e. Fermentation including the addition of at least one exogenous microorganism of interest, f. Stabilization of the fermented product from the previous step, g. Optionally, filtration of the stabilized product.

[0042] The collection step aims to specifically select non-valued by-products rich in molecules of interest, said molecules of interest being preferentially fermentable carbons, polyphenols, proteins, peptides, lipids, beta glucans, alpha and beta bitter acids, organic acids and minerals.

[0043] According to a particular embodiment, the collection aims to select solid, liquid, or semi-liquid brewing by-products from brewing (e.g., spent grains, brewing purges), from fermentation (e.g., yeasts, purged hops, semi-liquid purges of filter media and yeasts). These by-products are particularly rich in sugars, dextrins, starch, trace elements, polyphenols (e.g., xanthohumol, catechins, ferulic acid), bitter acids (lupulone), proteins, peptides, beta-glucans, and minerals.

[0044] According to another embodiment, the collection aims to select cider by-products and by-products from apple juices such as apple pomace, rich in pectins, sugars and polyphenols (procyanidin, epicatechin).

[0045] According to another embodiment, the collection aims to select olive by-products from oil mills, particularly olive oil: olive mill wastewater and olive pomace, rich in pulp, vegetable cell wall, polyphenols (hydroxytyrosol).

[0046] According to another embodiment, the collection aims to select wine by-products from wineries or distilleries: grape seeds and stems rich in polyphenols (resveratrol).

[0047] According to another embodiment, the collection aims to select by-products of the chocolate industry, particularly cocoa pods rich in polyphenols.

[0048] According to another embodiment, the collection aims to select by-products of starch or potato industry, such as starch-rich vegetable waters and peels.

[0049] According to another embodiment, the collection aims to select by-products from fruit juice presses, particularly orange peels rich in polyphenols and essential oil.

[0050] According to another embodiment, the collection aims to select by-products of the dairy and cheese industry, particularly whey rich in lactic ferments and proteins (casein).

[0051] Thus, in the context of the invention, the process can be implemented from a relatively under-exploited or unvalued by-product, but also from a mixture of unvalued by-products.

[0052] Preferably, biowaste is chosen from brewery by-products, cider by-products, wine by-products, olive oil by-products, distillery by-products, cocoa by-products, coffee by-products, starch factory by-products, sugar factory by-products, fruit juice press by-products, and dairy and cheese industry by-products, and combinations thereof.

[0053] Even more preferably, the collected by-products are chosen from: • Brewing by-products are selected from trub (the brewing settling after boiling and decantation), fermentation yeast, hop purge, malt dregs, distillery wastewater, and mixtures thereof. • Wine by-products are selected from vinasse, lees, grape seeds, and mixtures thereof, • Distillery by-products are selected from trub, fermentation yeast, malt grains and mixtures thereof, • Cider by-products are selected from apple pomace, lees, and mixtures thereof, • Olive by-products are selected from olive mill wastewater, olive pomace, and mixtures thereof, • Cocoa by-products are selected from cocoa liquor, cocoa cake, and mixtures thereof, • The by-product of coffee is coffee grounds, • Starch by-products are selected from peels, wastewater, and mixtures thereof, • The by-products of sugar refineries are selected from molasses, vinasse, and mixtures thereof, • The by-products of fruit juices are selected from pomace, citrus peels, and mixtures thereof; and • their combination.

[0054] Thus, according to one variant, the process can be implemented from a mixture of by-products, for example brewing and olive oil by-products or wine and brewing by-products, or wine, olive oil and cider by-products.

[0055] According to this embodiment, the process according to the invention preferably comprises a step of collecting the by-products of interest directly followed by a step of mixing the by-products specifically selected in the preceding step, that is to say, before the physical separation step. According to another variant, the step of mixing the specifically selected by-products can be carried out subsequently, advantageously, after the physical separation step.

[0056] The process according to the invention therefore comprises, according to one object, a separation step, which aims to separate the different phases or components of the biowaste. Advantageously, this step aims to obtain a liquid fraction and a solid fraction. Advantageously, biosolution products, in particular biocontrol and biostimulation products, are intended to be sprayed in liquid form; it is therefore necessary to separate the solid components from the liquid in order to avoid leaving suspended particles that could clog the sprayer nozzles and affect the quality of the product application and consequently its effectiveness. Preferably, the liquid fraction is recovered. According to an alternative embodiment, the solid fraction can also be recovered.

[0057] Also, for by-products intended for fermentation, the solid phase is separated from the liquid phase before the fermentation step. For by-products intended for the extraction of molecules of interest, the solid phase is separated from the liquid phase after the extraction step.

[0058] Thus, preferably, when the process includes only a fermentation step, it includes a step of separation of the solid fraction and the liquid fraction, then a recovery of the liquid fraction, finally, the fermentation step is implemented.

[0059] According to another preferred embodiment, when the process includes only an extraction step, it includes the extraction step followed by a step of separation of the solid and liquid fraction and recovery of the liquid fraction.

[0060] According to another particularly preferred object, the separation step is carried out using a decanter-centrifuge. Its operating principle is advantageous in the context of the invention. Indeed, the centrifugation is implemented with a horizontal rather than vertical axis of rotation, making it possible to achieve a fairly high dry matter content and thus optimize the separation by limiting efficiency losses. solid / liquid separation. As an example, a purge of beer yeast harvested at 20% dry matter exits the decanter-centrifuge with 85% dry matter.

[0061] Once the separation step is completed, and therefore the liquid fraction is separated from the solid fraction (dry matter), said liquid fraction comprises fermentable sugars, polyphenols and other molecules of interest used for the extraction and fermentation step to obtain biosolution products, including biocontrol and biostimulation products.

[0062] According to another object, the separated solid containing starch, cellulose, and yeasts, depending on the type of by-product, can be used in a process for manufacturing probiotic-type feed and feed supplements for animal nutrition and health. For example, brewer's yeast concentrated to 85% dry matter using a decanter-centrifuge can be packaged using a pellet press or an extruder. According to a preferred embodiment, the fermentable solid can be inoculated with a microorganism of probiotic interest for animal health (for example, a yeast, a lactobacillus, or a bacillus), fermented, and packaged after drying using a pellet press or an extruder.

[0063] For example, when solid probiotics are intended for animal feed (brewer's yeast, probiotic supplement with lactobacilli), the product is transformed into granules using an extruder to improve shelf life (reduced moisture content following extrusion).

[0064] When the liquid fraction is separated and recovered, an extraction step of the molecules of interest present in said liquid fraction is implemented and / or a fermentation step.

[0065] Preferably, the extraction of the molecules of interest is carried out using green chemistry, that is to say, without toxic substances, for example petrochemical solvents, or substances known to have undesirable effects and, more generally, an impact on the environment and human health. Thus, the extraction is preferably implemented using natural substances with a low impact on environmental quality and human health.

[0066] According to a preferred object of the invention, the extraction step comprises the following substeps: a. cold maceration with an alcoholic solvent derived from natural fermentation, b. decoction, and c. possibly, a vacuum extraction step or by ultrasound or microwave.

[0067] Thus, maceration with an alcohol naturally present in the by-product allows the extraction of the organic molecules of interest. Finally, decoction, providing The heat helps to accelerate reactions, particularly the isomerization of molecules of interest, thus increasing the efficiency of the biosolution.

[0068] According to a variant of the invention, the extraction may further include an extraction step carried out by ultrasound, microwave, under vacuum or under nitrogen in order to further increase extraction yields, to extract oxidation-sensitive molecules and thus limit the oxidation phenomenon.

[0069] According to a specific example, when the process aims to transform a brewing by-product into a biostimulant product, the liquid by-product is heated to 80°C during a decoction step with stirring and the incorporation of hop purges. The polyphenols and bitter acids from the hops isomerize and dissolve in the decoction. This process is then catalyzed by the heat and the presence of alcohol and lasts for a minimum of 3 hours in order to gradually evaporate the alcohol and achieve a near-zero alcohol concentration. Furthermore, this process advantageously offers, as an indirect effect, the sterilization of the liquid fraction and its concentration of bitter acids, polyphenols, and molecules of interest.

[0070] In the context of the invention, as a replacement for the extraction step or when the extraction step is completed, the process includes a fermentation step comprising the addition of at least one exogenous microorganism of interest in order to use the fermentable carbons contained in the liquid fraction to propagate said exogenous microorganisms which themselves generate organic metabolites of interest.

[0071] Fermentation conditions depend on the microorganism used. Thus, a person skilled in the art, based on their general knowledge, can adapt the fermentation conditions according to the selected microorganism. For example, Bacillus subtilis is a facultative anaerobic bacterium and requires specific conditions for successful sporulation. The sporulation conditions for the Bacillus genus of bacteria involve a decrease in nutrient bioavailability and temperature and pH conditions less favorable to its survival (below 15°C, above 40°C, pH < 2, pH > 8).

[0072] According to another example, a Lactobacillus plantarum must be grown anaerobically and does not keep as well because it does not form spores.

[0073] Consequently, key fermentation parameters are monitored, including pH, temperature, microbial growth, population, viability and the concentration of metabolites of interest such as lactic acid, enzymes, growth hormones, lipopeptide, surfactant molecules etc.

[0074] According to a preferred object of the invention, to improve the yield, the fermentation is carried out at a temperature between 17 and 37°C and / or a pH between 3 and 7, more preferably between 5 and 6.

[0075] The fermentation step advantageously allows the conversion of biowaste or agri-food by-products into new biosolution products, whether a biocontrol agent, a biostimulant, a biofertilizer, or a probiotic containing their own active ingredients. To this end, the inventor has advantageously isolated and selected strains of exogenous microorganisms with fermentation capacities suited to the culture medium, i.e., the liquid or solid fraction, and capable of producing metabolites of interest depending on the desired final product.

[0076] To achieve this, the exogenous microorganism of interest is preferentially chosen from among a lactic acid ferment, a bacterium of a genus chosen from Bacillus, Lactobacillus, Pseudomonas, Acetobacter, Nitrobacter, Nitrosomonas, Rhizobium, Agrobacterium, and Streptomyces, a fungus of the genus Trichoderma, a yeast of the genus Saccharomyces, or Aureobasidium, and their combinations.

[0077] More preferably, the inventor was interested, without limitation, in the bacteria Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus pumilus for their ability to produce lipopeptides of the type fengycin, iturine, surfactin, lichenysin as well as in Trichoderma (Trichoderma harzianum, Trichoderma viride) and yeasts (kingdom of fungi) for their ability to secrete toxins against fungal pathogens of plants and initiate competition for nutrients in the environment to be colonized.

[0078] For strains intended for the manufacture of biostimulants and activators of composts, manures, slurries, the inventor was particularly interested, without limitation, in lactic ferments, in particular the species: Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus helveticus, Lactococcus lactis, Propionibacterium freudenreichii, the yeasts Saccharomyces cerevisiae and nitrifying bacteria such as Nitrosomonas and Nitrobacter.

[0079] When biowaste, organic effluents or agri-food by-products are rich in simple and complex sugars, these are preferentially fermented by Saccharomyces yeasts or Bacillus, Lactobacillus lactic acid bacteria.

[0080] When biowaste, organic effluents or agri-food by-products contain alcohol, they are preferentially fermented by Acetobacter acetic bacteria.

[0081] When biowaste or agri-food by-products are predominantly solid in composition (low moisture content) and rich in cellulose, they are preferentially transformed by Trichoderma, Aureobasidium fungi.

[0082] When biowaste, organic effluents or agri-food by-products are rich in polyphenols, organic acids that are difficult to ferment, these- these are preferentially transformed by Streptomyces bacteria (hydrolysis), Pseudomonas (oxidative pathway).

[0083] More preferably, the biowaste and microorganism pairs are described in Table 1 below.

[0084] [Tables 1] Industry By-product / biowaste pH initial Exogenous microorganism Brewing Hops wort / dregs 5-6 Bacillus: Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus , Bacillus methylotrophicus, Paenibacillus polymyxa; Lactobacillus: Lactobacillus plantaru m, Lactobacillus acidophilus, Lactobacillus helveticus; Lactococcus lactis; Saccharomyces cerevisiae; Acetobacter: Acetobacter aceti; Trichoderma: Trichoderma harzianum, Trichoderma viride Brewing yeast fermentation 4-5 Bacillus: Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus , Bacillus methylotrophicus, Paenibacillus polymyxa; Lactobacillus: Lactobacillus plantaru m, Lactobacillus acidophilus, Lactobacillus helveticus; Lactococcus lactis; Saccharomyces cerevisiae; Acetobacter: Acetobacter aceti;Trichoderma : Trichoderma harzianum, Trichoderma viride brassicole purge hop fermentation 4-5 Bacillus : Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus , Bacillus methylotrophicus, Paenibacillus polymyxa ; Lactobacillus : Lactobacillus plantaru m, Lactobacillus acidophilus, Lactobacillus helveticus ; Lactococcus lactis ; Saccharomyces cerevisiae ; Acetobacter : Acetobacter aceti ; Trichoderma : Trichoderma harzianum, Trichoderma viride brassicole malt dreche 5-6 Bacillus : Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus ; , Bacillus methylotrophicus, Paenibacillus polymyxa ; Lactobacillus : Lactobacillus plantant m, Lactobacillus acidophilus, Lactobacillus helveticus ; Lactococcus lactis ; Saccharomyces cerevisiae ; Acetobacter : Acetobacter aceti ; Tr ichoderma : Trichoderma harzianum, Trichoderma viride vinicole vinasse 3-4 Streptomyces : Streptomyces violatus, Pseudomonas : Pseudomonas fluorescens, Nitrosomon as : Nitrosomonas europaea Nitrobacter : Nitr obacter winogradskyi, Nitrobacter hamburgen sis, Rhizobium : Brady rhizobium japonicum, R hizobium leguminosarum, ;Agrobacterium: Agrobacterium tumefaciens, Aureobasidium: Aureobasidium pullulons winemaking (fermentation yeast) 4-5 Bacillus, Lactobacillus, Saccharomyces, Acetobacter, Trichoderma winemaking grape seeds 5-6 Streptomyces: Streptomyces violatus, Pseudomonas: Pseudomonas fluorescens, Nitrosomon as: Nitrosomonas europaea Nitrobacter: Nitrobacter winogradskyi, Nitrobacter hamburgensis, Rhizobium: Bradyrhizobium japonicum, Rhizobium leguminosarum, ; Agrobacterium: Agrobacterium tumefaciens, Aureobasidium: Aureobasidium pullulons distillery trub 5-6 Bacillus: Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus, Bacillus methylotrophicus, Paenibacillus pol ymyxa ; Lactobacillus: Lactobacillus plantaru m, Lactobacillus acidophilus, Lactobacillus helveticus ; Lactococcus lactis ; Saccharomyces cerevisiae ; Acetobacter: Acetobacter aceti ;Trichoderma: Trichoderma harzianum, Trichoderma viride distillery yeast fermentation 5-6 Bacillus: Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus; , Bacillus methylotrophicus, Paenibacillus polymyxa ; Lactobacillus : Lactobacillus plantant m, Lactobacillus acidophilus, Lactobacillus helveticus ; Lactococcus lactis ; Saccharomyces cerevisiae ; Acetobacter : Acetobacter aceti ; Tr ichoderma : Trichoderma harzianum, Trichoderma viride distillery malt dregs 5-6 Bacillus : Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus , Bacillus methylotrophicus, Paenibacillus polymyxa ; Lactobacillus : Lactobacillus plantaru m, Lactobacillus acidophilus, Lactobacillus helveticus ; Lactococcus lactis ; Saccharomyces cerevisiae ; Acetobacter : Acetobacter aceti ; Tr ichoderma : Trichoderma harzianum, Trichoderma viride cidricole apple marc 4-5 Bacillus : Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus , Bacillus methylotrophicus, Paenibacillus polymyxa ; Lactobacillus : Lactobacillus plantaru m, Lactobacillus acidophilus, Lactobacillus helveticus;Lactococcus lactis; Saccharomyces cerevisiae; Acetobacter: Acetobacter aceti; Tr ichoderma: Trichoderma harzianum, Trichoderma viride cidricole lie (yeast fermentation) 4-5 Bacillus: Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus, Bacillus methylotrophicus, Paenibacillus polymyxa; Lactobacillus: Lactobacillus plantaru m, Lactobacillus acidophilus, Lactobacillus helveticus; Lactococcus lactis; Saccharomyces cerevisiae; Acetobacter: Acetobacter aceti; Tr ichoderma: Trichoderma harzianum, Trichoderma viride oléicole margine 4-5 Streptomyces: Streptomyces violatus, Pseudomonas: Pseudomonas fluorescens, Nitrosomon as: Nitrosomonas europaea Nitrobacter: Nitr ; obacter winogradskyi, Nitrobacter hamburgen sis, Rhizobium : Bradyrhizobium japonicum, R hizobium leguminosarum, ; Agrobacterium : Agrobacterium tumefaciens, Aureobasidium : A ureobasidium pullulons olive oil grignon 4-5 Streptomyces : Streptomyces violatus, Pseudo monas : Pseudomonas fluorescens, Nitrosomon as : Nitrosomonas europaea Nitrobacter : Nitrobacter winogradskyi, Nitrobacter hamburgen sis, Rhizobium : Bradyrhizobium japonicum, R hizobium leguminosarum, ; Agrobacterium : Agrobacterium tumefaciens, Aureobasidium : A ureobasidium pullulans cacao liqueur de cacao 5-6 Streptomyces : Streptomyces violatus, Pseudo monas : Pseudomonas fluorescens, Nitrosomon as : Nitrosomonas europaea Nitrobacter : Nitrobacter winogradskyi, Nitrobacter hamburgen sis, Rhizobium : Bradyrhizobium japonicum, R hizobium leguminosarum, ;Agrobacterium : Agrobacterium tumefaciens, Aureobasidium : A ureobasidium pullulans cocoa cocoa cake 5-6 Streptomyces : Streptomyces violatus, Pseudomonas : Pseudomonas fluorescens, Nitrosomon as : Nitrosomonas europaea Nitrobacter : Nitrobacter winogradskyi, Nitrobacter hamburgen sis, Rhizobium : Bradyrhizobium japonicum, R hizobium leguminosarum, ; Agrobacterium : Agrobacterium tumefaciens, Aureobasidium : A ureobasidium pullulans coffee coffee grounds 6-7 Streptomyces : Streptomyces violatus, Pseudomonas : Pseudomonas fluorescens, Nitrosomon as : Nitrosomonas europaea Nitrobacter : Nitrobacter winogradskyi, Nitrobacter hamburgen sis, Rhizobium : Bradyrhizobium japonicum, R hizobium leguminosarum, ; Agrobacterium : A ; Grobacterium tumefaciens, Aureobasidium: Aureobasidium pullulons Potato peels 6-7 Bacillus: Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus, Bacillus methylotrophicus, Paenibacillus polymyxa; Lactobacillus: Lactobacillus plantaru m, Lactobacillus acidophilus, Lactobacillus helveticus; Lactococcus lactis; Saccharomyces cerevisiae; Acetobacter: Acetobacter aceti; Trichoderma: Trichoderma harzianum, Trichode ma viride Potato waste 6-7 Bacillus: Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus, Bacillus methylotrophicus, Paenibacillus polymyxa; Lactobacillus: Lactobacillus plantaru m, Lactobacillus acidophilus, Lactobacillus helveticus; Lactococcus lactis; Saccharomyces cerevisiae; Acetobacter: Acetobacter aceti;Tr ichoderma : Trichoderma harzianum, Trichoderma viride fruit juice citrus peels 5-6 Streptomyces : Streptomyces violatus, Pseudomonas : Pseudomonas fluorescens, Nitrosomon as : Nitrosomonas europaea Nitrobacter : Nitr obacter winogradskyi, Nitrobacter hamburgen sis, Rhizobium : Brady rhizobium japonicum, R hizobium leguminosarum, ; Agrobacterium : Agrobacterium tumefaciens, Aureobasidium : Aureobasidium pullulans fruit juice marc 5-6 Bacillus : Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus , Bacillus methylotrophicus, Paenibacillus pol ymyxa ; Lactobacillus : Lactobacillus plantaru m, Lactobacillus acidophilus, Lactobacillus helveticus ; Lactococcus lactis ; Saccharomyces cerevisiae ; Acetobacter : Acetobacter aceti ; Tr ; ichoderma : Trichoderma harzianum, Trichode ma viride beet molasses 6-7 Bacillus : Bacillus subtilis, Bacillus amyloliqu efaciens, Bacillus velezensis, Bacillus pumilus , Bacillus methylotrophicus, Paenibacillus pol ymyxa ; Lactobacillus : Lactobacillus plantaru m, Lactobacillus acidophilus, Lactobacillus helveticus ; Lactococcus lactis ; Saccharomyces cerevisiae ; Acetobacter : Acetobacter aceti ; T ichoderma : Trichoderma harzianum, Trichode ma viride beet vinasse 5-6 Streptomyces : Streptomyces violatus, Pseudomonas : Pseudomonas fluorescens, Nitrosomon as : Nitrosomonas europaea Nitrobacter : Nitr obacter winogradskyi, Nitrobacter hamburgen sis, Rhizobium : Bradyrhizobium japonicum, R hizobium leguminosarum, ; Agrobacterium : Agrobacterium tumefaciens, Aureobasidium : A ureobasidium pullulons

[0085] Finally, the process according to the invention includes a stabilization step for the extracted and / or fermented product. Preferably, this stabilization step is carried out either hot by heat treatment or cold by microfiltration, UV filtration, or acid treatment. More preferably, the heat treatment is hot stabilization between 75 and 80°C, or microfiltration possibly supplemented by UV filtration, and possibly further supplemented by acid treatment, which is a dosage selected from acetic, peracetic, propionic, and lactic acids. Stabilization notably ensures the inactivation or elimination of undesirable microorganisms for reasons of quality compliance with specifications, marketing authorization standards, and guarantees the product's shelf life.

[0086] Once the stabilization step is complete, a first form of the biosolution is obtained. This can be packaged, for example in 5L, 10L, 20L drums, 1000L IBCs for liquid products, and in containers of 1 to 25kg for solid products, depending on the distribution channel and the knowledge of a person skilled in the art.

[0087] Optionally, a filtration step may be implemented before packaging, particularly for liquid products intended to be sprayed. For this purpose, the product is filtered using a decanter-centrifuge followed by a mesh screen. A 200 µm filter is used to ensure efficient separation of the liquid fraction from any remaining solid particles. Advantageously, the filtration is carried out in continuous flow.

[0088] In the context of the invention, biosolutions are preferably chosen from biocontrol products, biostimulation products, technological additives, biological silage preservatives, zootechnical additives, agronomic additives, biofertilizers, agri-food preservatives, post-harvest preservatives or plant protection products, and probiotics.

[0089] Biostimulants stimulate the plant's natural defenses and also allow farmers to reduce their reliance on synthetic or fossil-based fertilizers. Furthermore, they enable the development of self-sufficient fertilization through the composting of manure, slurry, green waste, and cover crops. This is achieved by solubilizing nutrients linked to the organic or mineral components of the clay-humus complex, such as phosphorus and potassium, and by improving bacterial and fungal symbiosis with the plant, such as root mycorrhization, thanks to a more extensive soil microbial biodiversity. Biostimulants thus provide microorganisms and nutrients of interest to increase the efficiency of composting, humification, mineralization, mycorrhization, bacterial symbiosis, and the development of the soil's bacterial microflora.

[0090] Biocontrol products address environmental challenges (Ecophyto plan) and aim to replace synthetic molecules from petrochemicals with molecules from green chemistry or natural substances, with microorganisms secreting metabolites for phytosanitary purposes or competing for colonization of the environment against pathogens, with macro-organisms competing for colonization of the environment against pathogens or predators of these same pathogens with a lesser impact on environmental quality and human health.

[0091] Microorganism complexes are technological additives for composting and sanitizing livestock bedding, usable in organic farming for manure, slurry, green waste, and digestate. They contain essential minerals and natural plant matter fermented with selected microorganisms. They improve the efficiency of farmyard manure, digestate, and compost, and rapidly activate their biological transformation into humus. Specifically, by acidifying the pH and providing fermentable carbon, they modify the nitrogen structure of organic matter, lowering the C / N ratio and significantly reducing ammonia nitrogen volatilization. This improves the utilization of composted manure or slurry in relation to spreading specifications. The composting additive nitrifies and humifies manure and slurry, and reduces ammonia nitrogen losses through volatilization. and by leaching. This improves the efficiency of fertilization of livestock effluents and improves the sanitary conditions of livestock from a microbiological point of view (less diarrhea, mastitis, pathogenic cells in milk) and respiratory problems related to ammonia odors for workers and livestock (e.g. lambs).

[0092] Finally, the manures, slurries, and droppings composted using our solution meet the NFU44-051 organic amendment standard, authorizing their commercialization. We thus move from a waste product to a commercial product with a value equivalent to synthetic fertilizer and a source of profit for the farmer.

[0093] Biological silage preservatives are technological additives that improve the lactic acid fermentation of forages during the ensiling process. They are produced from the fermentation of a plant by-product using selected homofermentative lactobacilli lactic acid bacteria. Stabilized at pH 4.2, they contain 1 to 2% residual sugars, which allows for the rapid acidification of the forage to be ensiled and provides sugars for low-nutrient forages such as grass, thus quickly triggering lactic acid fermentation. The benefit is better preservation of nutritional qualities over time and even improvement of the nutritional qualities of the ensiled forage, as well as protection against the development of molds, clostridia, and the secretion of mycotoxins.

[0094] Finally, probiotics for animal feed are dietary supplements containing live organisms, that is, microorganisms that will influence the health of the animal ingesting the probiotic, primarily at the level of the microbiota. Certain yeasts and bacteria (in our case, brewer's yeast and lactobacilli) improve digestive efficiency, particularly in ruminants, through their ability to absorb oxygen from the rumen and thus promote the activity of anaerobic bacteria. Studies have shown an improvement in average daily weight gain of 2.5 to 5% and an improvement in feed conversion ratio of 2% (weight gain / weight of feed consumed).

[0095] According to another aspect, the invention also relates to biosolutions selected from biocontrol products, biostimulation products, technological additives, biological silage preservatives, zootechnical additives, agronomic additives, biofertilizers, agri-food preservatives, post-harvest preservatives or plant protection products, and probiotics obtained by the process according to any of the embodiments described above.

[0096] In a particularly preferred manner, the biosolution is a biostimulation product or a biofertilizer and the exogenous microorganism of interest is a bacterium belonging to the genus chosen from Lactobacillus, Bacillus, Trichoderma, Saccharomyces, Acetobacter, and their combinations.

[0097] According to another preferred embodiment, the biosolution is a biocontrol product and the microorganism is chosen from Bacillus, Trichoderma, Streptomyces, Pseudomonas, Aureobasidium, Saccharomyces, and their combinations.

[0098] According to another preferred embodiment, the biosolution is a zootechnical additive or a probiotic and the exogenous microorganism of interest is chosen from Bacillus, Lactobacillus, Saccharomyces and their combinations.

[0099] According to another preferred embodiment, the biosolution is an agri-food additive, or a silage preservative, or a food preservative and the exogenous microorganism of interest is chosen from Bacillus, Lactobacillus, Saccharomyces and their combinations.

[0100] Thus, the process according to the invention is low carbon, consumes little energy and external resources other than bio-waste or by-products from the agri-food industry and meets the needs of the prior art.

[0101] The invention is now illustrated by non-limiting examples of compositions according to the invention and by results. Examples

[0102] Example 1 - Plant defence biostimulant

[0103] This biostimulant product has the following characteristics: it is an extract of hop polyphenols and beta bitter acids also containing amino acids, peptides, proteins, and trace elements, which are elicitor molecules enabling the activation of plant defense genes, fungicidal and bactericidal activity, and antioxidant activity protecting against oxidative stress in the plant. It is intended for application to the foliage of vegetable crops, vines, orchards, field crops (cereals, oilseeds, legumes, sugar beets), green spaces (lawns), and in horticulture. This product is obtained, in particular, according to the following process.

[0104] Hop purging is collected from a brewery and reprocessed by a decoction method at a temperature of 80°C for 3 hours to extract the polyphenols and bitter acids of interest, optionally supplemented by vacuum extraction or ultrasonic or microwave extraction, followed by a separation step using a decanter-centrifuge to recover the liquid fraction. The separated liquid is then filtered at 200 µm. The biostimulant is stable at a pH between 4 and 4.5 before packaging. The minimum concentration of polyphenols, calculated as gallic acid, tannic acid, catechin, or epicatechin equivalents, is 500 mg / kg in the biostimulant.

[0105] Example 2 - Nutritional bio-fertilizer

[0106] This biofertilizer has the following characteristics: it is a fermented liquid solution from hop distillers' grains containing lactic acid bacteria and yeasts and is intended to be applied to the sowing (at ground level) of field crops (cereals, oilseeds, legumes, beetroot), market garden crops, in vineyards, arboriculture, green spaces and horticulture.

[0107] This product is obtained in particular according to the following process.

[0108] Hop spent grain is collected from a brewery and reprocessed using a decanter-centrifuge separation process. The separated liquid phase is then fermented with selected strains of Lactobacillus plantarum and Bacillus subtilis. Fermentation takes place over 7 days at a temperature of 20°C to 30°C, with an initial pH of 5 to 6 and a final pH of 4 to 5. The liquid product of the fermentation is filtered at 200 µm and stabilized at pH 4–4.5 before packaging. The minimum lactic acid concentration in the biofertilizer is 20 g / L. The lactic acid solubilizes, in particular, the phosphorus and potassium trapped in the clay-humus complex.

[0109] A variant of the biofertilizer would consist of freeze-drying the bacteria from the liquid in order to condition the product in powder form.

[0110] Example 3 - Fungicide biocontrol

[0111] This biocontrol product has the following characteristics: it is fermented hop distillers' grains through inoculation with a selected strain of Bacillus subtilis or Bacillus amyloliquefaciens capable of synthesizing surfactant fungicidal molecules (iturine, surfactin, fengycin) and is intended to be applied to vines, arboriculture, field crops (cereals, oilseeds, legumes, beetroot), market garden crops, green spaces and horticulture.

[0112] This product is obtained in particular according to the following process.

[0113] Hop spent grain is collected from a brewery and reprocessed using a decanter-centrifuge separation process. The separated liquid phase is then fermented with selected strains of Bacillus subtilis or Bacillus amyloliquefaciens. Fermentation takes place over 15 days at a temperature of 20°C to 30°C, with an initial pH of 5 to 6 and a final pH of 4 to 5. The liquid product from the fermentation is filtered at 200 µm and stabilized at pH 4–4.5 before packaging.

[0114] According to one variant, the biocontrol bacteria in liquid form are lyophilized and the biocontrol is then packaged in powder form.

[0115] Example 4 - Biocontrol of biting insects

[0116] This biocontrol product has the following characteristics: it is a maltodextrin extract (a European basic substance effective in controlling piercing-sucking insects) and is intended to be applied to the leaf surface of Market gardening, vineyards, arboriculture, field crops (cereals, oilseeds and protein crops, legumes, sugar beets), green spaces and horticulture. This product is obtained in particular using the following process:

[0117] Malted wastewater is collected from a brewery or whisky distillery and reprocessed by a decoction process up to a temperature of 80°C in order to concentrate the complex sugars and extract the maltodextrin from the starch, followed by a separation step using a decanter-centrifuge to recover the liquid fraction. The separated liquid is then filtered at 200 µm. The final product is stabilized at a pH between 4 and 4.5 before packaging.

[0118] Example 5 - Technological additives: complex of microorganisms

[0119] This technological additive has the following characteristics: it consists of fermented hop grains containing selected strains of Lactobacillus plantarum, Lactococcus lactis, Bacillus subtilis, and Saccharomyces cerevisiae. This complex of microorganisms is a livestock bedding sanitizer and manure activator. It is intended for application to livestock bedding, manure piles, slurry, and green waste to address health issues and transform manure into a commercial organic amendment in accordance with standard NFU 44-051.

[0120] This product is obtained in particular according to the following process:

[0121] Hop spent grain is collected and reprocessed using a decanter-centrifuge separation process. The separated liquid phase is then fermented with selected strains of Lactobacillus plantarum, Lactococcus lactis, Bacillus subtilis, and Saccharomyces cerevisiae. Fermentation takes place over 15 days at a temperature of 20°C to 30°C, with an initial pH of 5 to 6 and a final pH of 4 to 5. The liquid product from the fermentation is filtered at 200 µm and stabilized at pH 4–4.5 before packaging. The minimum lactic acid concentration in the final product is 20 g / L. In this context, the lactic acid prevents ammonia volatilization and combats pathogenic bacteria. This also contributes to the plant's health benefits and future fertilization quality.

[0122] A variant of the final product would consist of freeze-drying the bacteria from the liquid in order to package a final product in powder form.

[0123] Example 6 - Biological silage preservatives

[0124] This biological silage preservative has the following characteristics: it is fermented malted wastewater containing a selected strain of Lactobacillus plantarum and is intended to be applied to grass silage, alfalfa silage, corn silage in order to improve the lactic fermentation of the silage process and to improve the preservation of the silage against molds, clostridia, mycotoxins.

[0125] This product is obtained in particular according to the following process.

[0126] Malted wastewater is collected from a brewery or whisky distillery. It is treated by a decanter-centrifuge separation process, and the separated liquid phase is then fermented with selected strains of Lactobacillus plantarum. Fermentation takes place over 15 days at a temperature of 20°C to 30°C, with an initial pH of 5 to 6 and a final pH of 4 to 5. The liquid product from the fermentation is filtered at 200 µm and stabilized at pH 4–4.5 before packaging. The minimum lactic acid concentration in the final product is 20 g / L, sufficient to preserve the feed.

[0127] A variant of the final product would consist of freeze-drying the bacteria from the liquid in order to package a final product in powder form.

[0128] Example 7 - Ruminant probiotics

[0129] This probiotic has the following characteristics: it is fermented solid hop distillers' grains containing selected strains of Lactobacillus plantarum, Bacillus subtilis, Saccharomyces cerevisiae and is intended to be used as a probiotic nutritional supplement for animal feed in order to improve rumination performance, cure rumen acidosis, reduce diarrhea and improve digestion in animals and humans in general.

[0130] This product is obtained in particular according to the following process:

[0131] Hop spent grain is collected from a brewery and then fermented with selected strains of Lactobacillus plantarum, Bacillus subtilis, and Saccharomyces cerevisiae. Fermentation takes place over a period of 3 weeks at a temperature of 20°C to 30°C, with an initial pH of 5 to 6 and a final pH of 4 to 5. The fermentation product is further processed by a decanter-centrifuge separation method. The separated solid phase is then passed through an extruder or pellet press to be packaged in pellet form.

[0132] Effectiveness test of biosolutions according to the invention

[0133] Example 8 - In vitro efficacy test of biosolutions according to the invention

[0134] This test aims to evaluate the effectiveness of the biocontrol according to the invention with B. subtilis against the black-rot agent (Phyllostica ampellicida).

[0135] The protocol is as follows. The tests are performed in Petri dishes on synthetic Oat-Malt-Agar medium. Growth inhibition curves are plotted using different concentrations of the products to determine the IC50 and MIC (50% inhibiting concentration and minimum inhibitory concentration). The product to be tested is incorporated into the nutrient medium at a rate of 1 mL of product per Petri dish, with 4 dishes per dose of product. Inoculation of the pathogens (extemporaneously) is carried out by depositing three mycelial implants per Petri dish. Controls are performed by applying the same quantity of sterile distilled water. The size of the mycelial growth is measured after 10–14 days of incubation at 22 °C. The effectiveness of the treatment on black rot is determined according to the formula below.

[0136] [Math.l]

[0137] The results are presented in Table 2 below.

[0138] [Tables2] mo-dashtss. % snhdddsm SEM K £4 lœ.ÙO? s' SCS 0.12 S 1Û0.0G BC £20 O 5J8 P! 8.02 i S8.&& coi hs.os ego L4? 18.41 C75

[0139] From a dose-response curve, the MIC, mean inhibitory concentration (minimum concentration to have 100% growth inhibition) and FIC50, concentration inducing a 50% reduction in the growth of the pathogen, were calculated.

[0140] The results inducing 100, 80 and 50% inhibition of Black-rot growth in the in vitro tests are presented in Table 3 below.

[0141] [Tables3] x” debts: d® h fraction por® CMI 0.2ÎG 5 a» 0« 18 LfôÔ 1JS

[0142] This B. subtilis-based biocontrol agent demonstrates complete efficacy in these in vitro tests, reflecting a direct mode of action (antibiosis). This complete inhibition is achieved with a minimum 5-fold dilution of the stock solution.

[0143] Example 9 - Test _ of the effectiveness of the biostimulant according to example 1 and 4 and of the biocontrol according to example 3.

[0144] This test consists of evaluating the effectiveness of three biosolutions according to the invention against downy mildew of grapevines. The biosolutions were tested in combination with a reduced dose of 30% copper, namely the fungicide biocontrol according to Example 3 (M4), the biostimulant according to example 1 (M5) and the biostimulant according to example 4 (M6), compared with untreated control modalities (M1), full dose of copper alone (M2) and reduced dose of copper alone (M3).

[0145] Several treatment campaigns were carried out and described in Table 4 below

[0146] [Tables4] Treatments Dates Stages Dose Copper metal g / ha T1 21 / 04 / 2023 13,200 T2 28 / 04 / 2023 53,250 T3 04 / 05 / 2023 54,250 T4 10 / 05 / 2023 55,300 T5 17 / 05 / 2023 56,300 T6 25 / 05 / 2023 57,300 T7 02 / 06 / 2023 65,300 T8 08 / 06 / 2023 71,400 Total 2,300

[0147] The results are presented in [Fig.1].

[0148] The results of trials in vineyard plots conducted from April to July 2023 demonstrate superior efficacy of the biosolutions (treatments M4, M5, M6) compared to the full dose of copper (M2) and the reduced dose of copper alone (M3). Indeed, the average frequencies of downy mildew attack and intensity on leaves for the three biosolutions are lower than for the full and reduced doses of copper. This means that the three biosolutions evaluated (fungicide biocontrol, biostimulant according to example 1, and biostimulant according to example 4) have the capacity to replace at least 30% of the copper dose and provide additional efficacy in the control of downy mildew in grapevines.

Claims

Demands

1. A process for transforming biowaste and by-products from the agri-food industry into biosolutions intended to improve fertilization and / or crop protection, or animal health, said process comprising the following steps: a. Collection of biowaste b. Physical separation of the solid and liquid fractions of the collected biowaste c. Recovery of the liquid fraction d. Extraction of molecules of interest present in said liquid fraction e. Optionally, fermentation including the addition of at least one exogenous microorganism of interest and stabilization of the fermented product obtained f. Optionally, filtration of the stabilized product.

2. Processing method according to the preceding claim, characterized in that the separation step is carried out by means of a decanter-centrifuge.

3. A transformation process according to any one of the preceding claims, characterized in that the extraction of the molecules of interest is a green chemistry extraction.

4. Processing according to the preceding claim, characterized in that the extraction step comprises the following substeps: • cold maceration with alcoholic solvent from natural fermentation, • decoction, and • optionally, a vacuum extraction step or by ultrasound or microwave.

5. Processing method according to any one of the preceding claims, characterized in that the fermentation is carried out at a temperature between 17 and 37 °C and / or a pH between 3 and 7, preferably between 5 and 6.

6. A transformation process according to any one of the preceding claims, characterized in that the stabilization is carried out hot by heat treatment or cold by filtration and acid treatment.

7. Processing method according to the preceding claim, characterized in that the heat treatment is a hot stabilization between 75 and 80°C, or the acid treatment is a dosage selected from acetic, peracetic, propionic, and lactic acid.

8. A transformation process according to any one of the preceding claims, characterized in that the exogenous microorganism of interest is selected from a lactic acid ferment, a bacterium of a genus selected from Bacillus, Lactobacillus, Pseudomonas, Acetobacter, Nitrobacter, Nitrosomonas, Rhizobium, Agrobacterium, and Streptomyces, a fungus of the genus Trichoderma, a yeast of the genus Saccharomyces, or Aureobasidium; and combinations thereof.

9. A transformation process according to any one of the preceding claims, characterized in that the biowaste is selected from brewery by-products, cider by-products, wine by-products, olive oil by-products, distillery by-products, cocoa by-products, coffee by-products, starch factory by-products, sugar factory by-products, fruit juice press by-products, and dairy and cheese industry by-products, and combinations thereof.

10. A transformation process according to any one of the preceding claims, characterized in that the biosolution is selected from biocontrol products, biostimulation products, technological additives, biological silage preservatives, zootechnical additives, agronomic additives, biofertilizers, agri-food preservatives, post-harvest preservatives or plant protection products, and probiotics.

11. A transformation process according to any one of the preceding claims, characterized in that the biosolution is a biostimulation product or a biofertilizer and the exogenous microorganism of interest is a bacterium belonging to the genus selected from Lactobacillus, Bacillus, Trichoderma, Saccharomyces, Acetobacter, and their combinations.

12. A transformation process according to any one of the preceding claims, characterized in that the biosolution is a biocontrol product and the microorganism is selected from Bacillus, Trichoderma, Streptomyces, Pseudomonas, Aureobasidium, Saccharomyces, and their combinations.

13. A transformation process according to any one of the preceding claims, characterized in that the biosolution is a zootechnical additive or a probiotic and the exogenous microorganism of interest is chosen from Bacillus, Lactobacillus, Saccharomyces and their combinations.

14. Processing according to any one of the preceding claims, characterized in that the biosolution is an agri-food additive, or a silage preservative, or a food preservative and the exogenous microorganism of interest is chosen from Bacillus, Lactobacillus, Saccharomyces and their combinations.