Using insect pupariums, dead flies or mixtures thereof, and frass to increase plant growth

Abstract

Use of insect puparia, dead flies or mixtures thereof, and frass to increase plant growth. The present invention relates to the synergistic use of a combination of holometabolous insect puparia, or whole dead flies or their fragments, or a mixture thereof, and animal slurry, to accelerate plant growth and / or increase crop yield. It also relates to a method of cultivating a plant with these compositions. Figure for abstract: none

Description

Title of the invention: Use of insect puparia, dead flies or mixtures thereof, and frass to increase plant growth

[0001] The present invention relates to the use of a combination of a first composition (A) comprising (i) holometabolous insect puparia, (ii) whole dead flies or fragments thereof, or (iii) a mixture thereof, and a second composition (B) of animal slurry, for accelerating plant growth and / or for increasing crop yield. It also relates to a method of cultivating a plant with these compositions.

[0002] The search for new strategies aimed at maintaining or increasing the productivity of agricultural systems, while reducing the inputs of agricultural inputs, is of constant interest.

[0003] Among the alternative solutions to chemical compounds, chitin and its derivatives, such as chitosan (particularly derived from insects or crustaceans), are known as biopesticides or for stimulating the natural defenses of plants or their growth. These products are generally applied by foliar application or by mixing into the soil.

[0004] However, such products must first be extracted, transformed and / or purified, which is complex and time-consuming to implement.

[0005] Furthermore, rapid crop start-up is critical to the success of the crop, as it ensures good establishment. Good establishment limits the risk of destruction by pests, or even by difficult climatic conditions, and accelerates the initial growth of the plant. This generally ultimately allows for a better harvest yield and / or better resilience of the plant to external attacks.

[0006] There is therefore a need for alternatives to chemical inputs that are easy to apply and effective. In particular, there is a need for bio-sourced alternatives, particularly recycled ones. In particular, there is a need for a bio-sourced alternative that is easy and quick to obtain and less expensive. Said alternative is in particular a means of recovering a co-product (i.e. waste) generated in an industry.

[0007] The Applicant has now discovered, as shown in the examples, that the application of ground black soldier fly puparium (Hermetia illucens, or BSF, for black soldier fly) combined with the application of frass from this same fly, makes it possible to very significantly increase the growth of the aerial biomass and the root biomass of a cereal, such as corn, at least 54 days after sowing, compared to an untreated control. This increase is even of the order of synergy. The application of a ground material of black soldier fly puparia (Hermetia illucens) combined with the application of frass from this same fly, therefore presents a synergistic effect on plant growth and / or crop yield.

[0008] Puparium grind and frass are very easy and quick to obtain, bio-sourced, and allow the recycling of BSF breeding co-products.

[0009] The present invention therefore relates to the synergistic use (A) of a first composition comprising (i) holometabolous insect puparia, (ii) whole dead flies or their fragments, or (iii) one of their mixtures, and (B) of a second composition comprising animal manure, to accelerate plant growth and / or to increase crop yield.

[0010] The present invention also relates to the combined use of holometabolous insect puparia, or of whole dead flies or their fragments, or of one of their mixtures, and of animal slurry, to accelerate the growth of plants and / or to increase the crop yield.

[0011] This use according to the invention is a synergistic or combined use (hereinafter "combined use according to the invention") (A) of a first composition comprising (i) puparia of holometabolous insect, preferably of Hermetia illucens, (ii) flies, preferably Hermetia illucens, dead whole or their fragments, or (iii) one of their mixtures, and (B) of a second composition comprising animal manure, preferably frass of Hermetia illucens. The first composition (composition A) and the second composition (composition B) are generally two separate compositions, each applied according to specific methods detailed below. Alternatively, compositions (A) and (B) are premixed before use.

[0012] Animal manure, such as Hermetia illucens frass, may intrinsically comprise holometabolous insect puparia. However, the present invention aims at an exogenous supply of such puparia via the first composition (A). In other words, compositions (A) and (B) are different from each other.

[0013] The invention also relates to a method of growing a plant on a substrate, comprising: a. mixtures of the substrate with (A) a first composition comprising (i) puparia of holometabolous insect, preferably of Hermetia illucens, (ii) flies, preferably Hermetia illucens, dead whole or their fragments, or (iii) one of their mixtures, and with (B) a second composition comprising animal manure, preferably frass of Hermetia illucens, the mixtures of (A) and (B) being made simultaneously with the substrate, or in the order (A) then (B), or in the order (B) then (A); b. preparing a seedling with the substrate obtained in step a); then c. growing the seedling from step b).

[0014] By "plants" is preferably meant cereals or market garden plants. Preferably, the cereals are chosen from corn, wheat, rice, barley, millets (including sorghum), oats and rye. Preferably, the cereals are chosen from corn and wheat. Preferably, the market garden plants are chosen from salads such as lettuce, potatoes, green beans, cucumbers, spinach and carrots.

[0015] By "acceleration of plant growth" is meant an increase in the aerial (i.e. leaves, stems and flowers) and root biomass of at least 15%, preferably at least 20%, preferably at least 25%, compared to the same plant species treated with a control, after at least 54 days of cultivation. Preferably, the combined use according to the invention an increase in the aerial biomass (i.e. leaves, stems and flowers) of at least 10%, preferably at least 15%, preferably at least 20%, preferably at least 25%, compared to the same plant species treated with a control, after at least 15 days of cultivation, preferably at least 30 days of cultivation, preferably at least 54 days of cultivation.Preferably, the combined use according to the invention results in an increase in root biomass of at least 15%, preferably at least 20%, preferably at least 30%, preferably at least 35%, compared to the same plant species treated with a control, after at least 15 days of cultivation, preferably at least 30 days of cultivation, preferably at least 54 days of cultivation. The control is preferably the same animal manure as that present in the combined use according to the invention, but used alone. Preferably, the control is Hermetia illucens frass.

[0016] By "increasing the crop yield" is meant a significant increase in this yield. This yield can be measured by harvesting the biomass. Preferably, by "increasing the crop yield" is meant that the above-ground biomass (i.e. leaves, stems and flowers) of the crop is increased by at least 10%, preferably at least 15%, preferably at least 20%, preferably at least 22%, compared to the same crop treated with a control, after at least 15 days of cultivation, preferably at least 30 days of cultivation, preferably at least 54 days of cultivation. The control is preferably the same animal manure as that present in the combined use according to the invention, but used alone. Preferably, the control is Hermetia illucens frass.

[0017] The combined use according to the invention firstly comprises the use of a first composition comprising (i) puparia of holometabolous insect, preferably of Hermetia illucens, (ii) flies, preferably Hermetia illucens, dead whole or their fragments, or (iii) one of their mixtures.

[0018] According to a first embodiment, the first composition according to the invention comprises (i) holometabolous insect puparia. Holometabola are a superorder of insects. The development of holometabolous insects is characterized by a complete metamorphosis, i.e. an egg, then a larva (which differs greatly from adults), then a nymph, and finally an imago (adult). Preferably, the holometabolous insect is chosen from Diptera, preferably from Brachycera, preferably from Stratiomyidae.

[0019] Preferably, the holometabolous insect is the black soldier fly (Hermetia illucens, or BSF, for black soldier fly).

[0020] The black soldier fly (Hermetia illucens, or BSF, for black soldier fly) is a fly of the order Diptera, of the family Stratiomyidae. The life cycle of this insect is composed of 9 stages, namely: the egg, six larval stages, the pupa and the adult.

[0021] The cycle begins with an egg, oval in shape, about 1 mm long and pale yellow or creamy white in color. This egg hatches after about 4 days, producing a larva.

[0022] Newly hatched larvae measure about 1.8 mm. At the end of their growth, they are broad and oval and can reach 27 mm long and 6 mm wide. The larvae pass through six larval stages (L1 to L6), which can last from 15 days to 5 months (depending on conditions): the larval stages are differentiated by the size of the larvae, their color (which darkens as the stages progress) and the development of the setae. Between each larval stage, a moult occurs; the larva leaves the integument (exuvia) which has become too small, this rejection is called exuviation. The sixth larval stage is the prepupa stage. During this stage, the larvae turn reddish brown.

[0023] Then the prepupae become immobilized and begin their metamorphosis; we then speak of pupae. The pupa differs from the two other nymphal forms in that the metamorphosis takes place during a process comprising all the stages of moulting except exuviation. The individual is then completely enclosed in the last larval exuvia, called puparium, and completely immobile. This puparium is completely rigid due to quinone tanning of the free amine function of the structural proteins, which makes the exocuticle harder. The procuticle (i.e. exo- and endocuticles) of the puparium being detached from the future imago (adult), the whole takes on an ovoid shape.

[0024] After two to three weeks in the pupal stage, adults emerge (emergence of the imago) and reproduce when they are sexually mature (2 days after emergence). Upon emergence, the imago extracts itself from the puparium.

[0025] The cuticle, or exoskeleton, is the outer layer secreted by the epidermis of insects, here Hermetia illucens. It is generally formed of three layers:

[0026] - the epicuticle, which is the thinnest and outermost layer of the cuticle (less than 4 pm); this layer is impermeable to water and includes a waterproofing wax layer, as well as proteins and chitin, in smaller quantities;

[0027] - the exocuticle, which is the intermediate layer of the cuticle; it is composed of essentially hardened proteins, the tannins, which are responsible for the rigidity of the cuticle, chitin and possibly melanin; and

[0028] - the endocuticle, which is a thin, flexible layer, consisting of a mixture of proteins and chitin.

[0029] The exo- and endocuticles form the procuticle.

[0030] Preferably, according to a first embodiment, the first composition according to the invention comprises (i) puparia of holometabolous insect, preferably of Hermetia illucens.

[0031] By puparium of holometabolous insect, preferably of Hermetia illucens, is meant an empty puparium, i.e. after the imago has emerged. These pupariums are recovered after the emergence of adult flies; typically they are co-products of BSF rearing. The puparium is the envelope of the pupa. This envelope is rigid. The puparium is different from larval exuviae, which are the exuviae of the first six larval stages.

[0032] The puparia of Hermetia illucens generally have the following profile, in dry matter weight relative to the total weight of puparium: - a content of less than or equal to 25% by weight of chitin; - a content of at least 5% by weight of total fat.

[0033] Furthermore, the puparia of Hermetia illucens generally contain manganese at a level of at least 200 ppm, preferably at least 400 ppm (on dry matter). This manganese concentration is much higher than the larval exuviae. Indeed, for comparison, the larval exuviae typically contain manganese at a level of less than 110 ppm on dry matter. This manganese, in association with the other constituents of the puparia, seems to play a role in the observed increase in plant growth.

[0034] Preferably, the pupariums of Hermetia illucens have the following profile, in dry matter: - from 5% to 25% by weight of chitin relative to the total weight of puparium; - from 5% to 30% by weight of total fats relative to the total weight of puparium; and - from 200 ppm to 2000 ppm of manganese.

[0035] Preferably, the pupariums of Hermetia illucens have the following profile, in dry matter: - from 10% to 25% by weight of chitin relative to the total weight of puparium; - from 5% to 30% by weight, preferably from 5% to 20% by weight, of total fat relative to the total weight of puparium; and - from 200 ppm to 2000 ppm of manganese, preferably from 400 ppm to 1500 ppm, preferably from 600 ppm to 1000 ppm.

[0036] The interest lies on the one hand in the simplicity of use (no pre-processing or extraction step is necessary), and on the other hand in the interesting effects obtained.

[0037] The first composition comprising holometabolous insect puparia, whole dead flies or fragments thereof, or mixtures thereof according to the invention (composition (A)) comprises said puparia, said whole dead flies or fragments thereof, or mixtures thereof, as active ingredient. Preferably, this composition preferably consists exclusively of holometabolous insect puparia, preferably of Hermetia illucens.

[0038] The first composition comprising puparia of a holometabolous insect, preferably of Hermetia illucens according to the invention, is preferably a powder, or a paste. Indeed, preferably, the puparia of a holometabolous insect, preferably of Hermetia illucens, are used in powder form, or in paste form. For example, they can be obtained by grinding, for example in a mortar and pestle; the composition is then a ground puparia. Such a powder (or ground) of puparia of a holometabolous insect, preferably of Hermetia illucens according to the invention, differs from commercial products by its composition, i.e. it contains all the constituents of the puparia, and not just chitin. For example, such a powder (or ground material) of holometabolous insect puparia, preferably of Hermetia illucens according to the invention, comprises fatty substances. Moreover, chitin is not the majority in the puparia powder.Indeed, the chitin content in such a powder is typically less than 25% by weight in dry matter, preferably less than 24% by weight in dry matter.

[0039] According to a second embodiment, the first composition comprises (ii) whole dead flies or their fragments. Preferably, the whole dead flies or their fragments are chosen from brachycera, preferably from Stratiomyidae. Preferably, the whole dead flies or their fragments are dead black soldier flies (Hermetia illucens) or their fragments.

[0040] These dead flies, or their fragments, are typically present as co-products of BSF rearing. They are present as such, because not all flies survive after the emergence of the imago. They can be recovered as is.

[0041] The dead Hermetia illucens flies or their fragments according to the invention generally have the following profile, in dry matter weight relative to the total weight of dead flies: - a content of less than or equal to 10% by weight of chitin; - a content of at least 25% by weight of total fat.

[0042] Furthermore, the dead flies according to the invention generally contain manganese in a content of less than 50 ppm (on dry matter).

[0043] Preferably, the dead Hermetia illucens flies or their fragments according to the invention have the following profile, in dry matter: - a content of less than or equal to 7% by weight of chitin relative to the total weight of dead flies; - from 27% to 40% by weight of total fat relative to the total weight of dead flies; and - from 5 ppm to 40 ppm of manganese.

[0044] According to a third embodiment, the first composition comprises (iii) a mixture of puparia of holometabolous insect, preferably of Hermetia illucens, and whole dead flies or their fragments.

[0045] Preferably, such a mixture comprises the puparia of holometabolous insects, preferably of Hermetia illucens, having the profile described above (ingredient (i)).

[0046] Preferably, such a mixture comprises whole dead flies or their fragments having the profile described above (ingredient (ii)).

[0047] The mixture can be made in any proportion of puparia to whole dead flies or their fragments.

[0048] Preferably, the first composition according to the invention comprises a mixture (iii) of Hermetia illucens puparia, and whole dead Hermetia illucens flies or their fragments. Such a mixture is a co-product of BSF breeding.

[0049] Preferably, the first composition according to the invention is used in an amount of at least 0.05%, preferably at least 0.1%, preferably at least 0.5% by weight relative to the weight of dry substrate. Preferably, the first composition according to the invention is used in an amount of between 0.05% and 5%, preferably between 0.1% and 3%, preferably between 0.5% and 2% by weight relative to the weight of dry substrate.

[0050] By "substrate" is meant any substrate suitable for growing plants, preferably chosen from earth, potting soil, peat, sand and mixtures thereof. Preferably, the substrate is potting soil, preferably a mixture of peat and sand.

[0051] The combined use according to the invention also comprises the use of a second composition comprising animal manure, preferably Hermetia illucens frass.

[0052] Animal slurry can be any slurry usable in agriculture. By "slurry" is meant a mixture of livestock excrement and water. The slurry can also contain bedding residues such as straw. The animal can be any animal livestock, preferably it is chosen from holometabolous insects, pigs, cattle and poultry. Preferably, the slurry is a holometabolous insect slurry. Preferably, the slurry is a Stratiomyidae slurry, preferably the slurry is Hermetia illucens frass.

[0053] Frass is an Anglo-Saxon word designating the mixture of excrement and scraping resulting from the feeding activity of certain insects, including here Hermetia illucens. Thus, by frass is meant the slurry &Hermetia illucens. In the context of the present invention, frass &Hermetia illucens designates a mixture which comprises excrement of BSF larvae, exuviae of larvae and other parts of other developmental stages of BSF (including dead eggs, larvae, pupae, puparia or adults), indigestible materials, for example fibrous materials or cellulose-based materials, other metabolic products, for example hormones, antibiotics or enzymes, chitin, and other organisms associated with this organic mixture, such as bacteria, fungi, protozoa and yeasts.Preferably, frass &Hermetia illucens is the result of a process in which the majority (>50%, up to 100%) of the dry matter of a raw material passes through the digestive system of BSF larvae to produce the residues that constitute frass.

[0054] According to the invention, the second composition comprising animal slurry, preferably frass & Hermetia illucens, is enriched with puparia by mixing with the first composition (composition (A)). This mixture allows an exogenous supply of puparia to the slurry such as frass.

[0055] According to the invention, animal manure, preferably frass & Hermetia illucens, has beneficial effects on plant growth and survival. Preferably, the frass of Hermetia illucens contains macronutrients, for example it has an NPK profile at levels around 3-3-3 (expressed in %).

[0056] As such, the animal manure, preferably frass & Hermetia illucens, may be used as a fertilizer or soil additive, for example at an application rate appropriate for the selected plant and soil characteristics. Preferably, the animal manure, preferably frass & Hermetia illucens, is used in an amount of at least 0.5 T / ha of substrate, preferably at least 3 T / ha. Preferably, the frass & Hermetia illucens is used in an amount of between 0.5 T / ha and 10 T / ha of substrate, preferably between 2 T / ha and 10 T / ha of substrate, preferably between 4 T / ha and 10 T / ha of substrate.

[0057] The second composition comprising animal slurry, preferably Hermetia illucens frass according to the invention, can be obtained by simple recovery / collection of the frass.

[0058] The combination of the first and second compositions according to the invention can be applied by any means. In particular, it can be applied directly to the growing medium, such as soil (earth), for example by irrigation; or applied to plants, for example to the roots, to aerial parts such as leaves, flowers and / or stems; or applied to seeds.

[0059] Preferably, the first composition according to the invention is premixed with the second composition, i.e. with the animal slurry, before use.

[0060] Alternatively, preferably, the first composition according to the invention is applied before the second composition, i.e. the animal slurry. Alternatively, preferably, the first composition according to the invention is applied after the second composition, i.e. the animal slurry.

[0061] In all cases, the application of the first and second compositions makes it possible to enrich the slurry, preferably the frass, with puparia.

[0062] Preferably, the first composition according to the invention is mixed with animal manure, preferably Hermetia illucens frass, then the resulting mixture is added directly to the substrate.

[0063] The invention also relates to a method of growing a plant on a substrate, comprising: a. mixtures of the substrate with (A) a first composition comprising (i) puparia of holometabolous insect, preferably of Hermetia illucens, (ii) flies, preferably Hermetia illucens, dead whole or their fragments, or (iii) one of their mixtures, and with (B) a second composition comprising animal manure, preferably frass of Hermetia illucens, the mixtures with (A) and (B) being carried out simultaneously with the substrate, or in the order (A) then (B), or in the order (B) then (A); b. preparing a seedbed with the substrate obtained in step a); then c. cultivation of the seed from step b).

[0064] The compositions (A) and (B) according to the invention, used in step a), are identical to those described above.

[0065] The method according to the invention thus comprises a first step (step a) of mixing a substrate with the first composition (A) and with the second composition (B).

[0066] According to a first embodiment, compositions (A) and (B) are premixed before mixing with the substrate. According to a second embodiment, compositions (A), (B) and the substrate are mixed at the same time. These first and second embodiments correspond to simultaneous mixing with the substrate.

[0067] According to a third embodiment, the composition (A) is first mixed with the substrate, then the mixture obtained is then mixed with the composition (B).

[0068] According to a fourth embodiment, composition (B) is first mixed with the substrate, then the mixture obtained is then mixed with composition (A).

[0069] At the end of step a), the plant culture substrate is mixed with (i) the holometabolous insect puparia, preferably of Hermetia illucens, (ii) the whole dead flies or their fragments, or (iii) a mixture thereof, and with the animal slurry, preferably the frass of Hermetia illucens.

[0070] This substrate is used to prepare a seedling of the plant to be grown: this is step b). For this purpose, typically, at least one seed of the plant is sown in the substrate. The preparation of the seedling is carried out in a conventional manner, for example by sowing the substrate, then covering with a layer of substrate, possibly moist.

[0071] The conditions for growing the seed, then the plant, are the classic conditions for the plant in question. At the end of step b), at least one seed is obtained on the substrate containing the puparia, the whole dead flies or their fragments, or their mixtures, and the animal manure, preferably the frass of Hermetia illucens.

[0072] Finally, step c) includes the cultivation of the seedling. The cultivation is also conventional, and is carried out under temperature and watering conditions adapted to each plant.

[0073] The following examples are given for illustrative purposes.

[0074] Example 1: Measurement of aerial biomass in corn plants treated with the combined use according to the invention

[0075] The crop studied is untreated corn of the DKC4162 variety at a rate of 3 seedlings per pot.

[0076] CULTURE SUBSTRATE:

[0077] The tests were carried out in 3 L growing pots. The surface area of ​​the pots was 283.52 cm2. The test substrate was a clod soil made up of peat with added sand (quartz sand) in a proportion of 70% soil / 30% sand w / w.

[0078] The characteristics of the soil were as follows:

[0079] - Composition: blond peat - black peat

[0080] - Retention capacity: 750 mL / L

[0081] - pH: 5.8

[0082] - Resistivity: 1900 Q / cm

[0083] - Organic matter: 30%.

[0084] CULTIVATION CONDITIONS:

[0085] All of the growing pots were filled with the same weight of substrate without manual compaction.

[0086] In each of the pots, 6 seeds were sown evenly on the surface. As soon as they emerged, the excess seedlings were pulled out to leave only 3 plants per pot. During the 15 days following emergence, the regrowth was monitored and systematically pulled out so as to keep only 3 plants per pot. The pots were then watered by sub-irrigation so as to saturate the substrate with water.

[0087] Following sowing, the plants were placed in a tunnel greenhouse under the following growing conditions:

[0088] - Lighting 16 h day / 8 h night

[0089] - Temperature: 15°- 30°C

[0090] - Brightness from 1500 to 2500 lux

[0091] Temperature and humidity monitoring was carried out using a recorder, the data were collected and attached to the trial report. The trial was block randomized. Untreated controls were included in the device. The plants were kept in culture for 54 days ~ 8 weeks.

[0092] FERTILIZATION:

[0093] Regarding the conventionally fertilized conditions (chemical fertilizer), fertilization was reduced in order to potentiate the effects of the tested products: it was reduced by 50% compared to the conventional fertilization program. Fertilization was divided into 2 applications: a first application at sowing and a second application at the 4 / 6 leaf stage in the growing medium. Depending on the response of the crop to the fertilization gradient, it was planned to modulate the nitrogen applications, particularly at the 2nd application.

[0094] Concerning the organic fertilizer NPK 3-3-3 (85-90% MS) composed of 100% Hermetia illucens frass and presented in the form of pellets of diameter 4mm, it was brought at 8 T / ha, the theoretical quantities of NPK were 240 U of N, 240 U of P2O5 and 240 U of K2O.

[0095] It was added in one go at the time of substrate preparation.

[0096] Preparation of puparium samples

[0097] Black soldier fly puparia (Hermetia illucens) were collected and ground in a mortar and pestle to obtain a powder.

[0098] 2 g of this powder are mixed in 15 ml of water to obtain a solution. then collects the supernatant.

[0099] WATER STRESS:

[0100] Water stress was a chronic stress. It was carried out according to the following procedure: reduction of watering by 50%, 11 days after the total emergence of the plants.

[0101] Chronic stress protocol:

[0102] After sowing, the pots were watered regularly to maintain optimum humidity for growth. Watering was carried out by sub-irrigation, by filling the cups with water until the substrate was saturated. After 2 hours, the cups were emptied.

[0103] 11 days after the plants had fully emerged, water stress started. It was a chronic water stress which consisted of a systematic decrease of the quantity of irrigation water until the end of the test.

[0104] After a final watering to saturation of the pots, 10 pots were weighed and an average of the value was calculated so as to obtain a weight which should represent the weight of the wet pot.

[0105] After 5 days, 10 pots were weighed, the average was then calculated. This value was to represent the weight of the dry pot.

[0106] The difference between the weight of the wet pot and the weight of the dry pot was calculated and represented the quantity of water consumed. This quantity was supplemented on all the non-stressed pots (only the non-stressed chemical fertilizer modality, condition 1) and half of this quantity was provided on all the stressed pots.

[0107] Example:

[0108] Wet pot weight = 3.5 kg

[0109] Dry pot weight = 3.250 kg

[0110] Quantity of water consumed = 3.5-3.250 = 250 g

[0111] Quantity of water to be added to unstressed pots = 250 g

[0112] Quantity of water to be added to stressed pots = 125 g.

[0113] In order to take into account the growth of the plants and their increasing need for water, this manipulation was repeated on the non-stressed pots after the start of the stress and the quantity of water to be provided was adapted. According to the same principle, the stressed pots received half the quantity of water of the non-stressed pots.

[0114] DOSES STUDIED:

[0115] Hermetia illucens frass and Hermetia illucens puparia were added to the substrate by mixing. The frass doses are calculated in T / ha (100% represents 8 T / ha) and the puparia doses in % of the dry substrate.

[0116] 18 modalities were tested: - From 1 to 7: 7 modalities: constant base 50% fertilization, chemical fertilizer + 7 distinct doses of pupariums, namely respectively 0%, 0.5%, 1%, 2.5%, 0.25%, 0.1% and 0.05% in mass of dry substrate (without water stress); - From 12 to 18: 7 modalities: constant base 50% fertilization, chemical fertilizer + 7 distinct doses of pupariums, namely respectively 0%, 0.5%, 1%, 2.5%, 0.25%, 0.1% and 0.05% in mass of dry substrate (with water stress); and - From 8 to 11: 4 modalities with 100% fertilization (8 and 9) or 90% fertilization (10 and 11), without water stress; different combinations of frass + doses of pupariums, namely: • 8: 8 T / ha + 0% pupariums (by mass of dry substrate), • 9: 8 T / ha + 0.5% pupariums (by mass of dry substrate), • 10: 7.2 T / ha + 0.5% pupariums (by mass of dry substrate), • 11: 7.2 T / ha + 1% pupariums (by mass of dry substrate).

[0117] The protocol was subject to an operating mode aimed at guaranteeing statistical representativeness (e.g. randomized blocks). For each modality, 5 repetitions (pots) were used. Results :

[0118] Biomasses are measured in grams (g).

[0119] The results show the following: A significant improvement in plant growth (measured by the quantity of aerial biomass at +54 days and +74 days after sowing) for the combined use according to the invention: • At +54 days (see Table 1): • For the same dose of frass (100%), an addition of 0.5% of puparium ground material improves performance by +10.4%; • For the same dose of frass (90%), an increase in the addition of 0.5% of puparium ground material (from 0.5% to 1.0%) improves performance by +20.4%; and • For a lower dose of frass (90% vs. 100%), an addition of 1.0% puparium ground improves performance by +28.5%.

[0120] [Tables 1] Modality Frass Puparium s Aerial biomass 54d vs modality 8 vs modality 10 8 100% 0% 259.4 9 100% 0.50% 286.4 10.4% 10 90% 0.50% 276.8 11 90% 1% 333.3 28.5% 20.4%

[0121] For modalities 2 to 7, the aerial biomass at 54 days varies between 326.9g and 350.4g. For modality 1 (chemical fertilizer alone), the aerial biomass at 54 days is 334.3g.

[0122] For modalities 12 to 18, the aerial biomass at 54 days varies between 214.4g and 227.8g.

[0123] The aerial biomass of modalities 9 to 11 (11 in particular) is comparable to that of modalities 2 to 7 (with chemical fertilizer instead of frass, and without water stress).

[0124] Thus, these results show that the performance of the puparium + chemical fertilizer combinations (modalities 2 to 7) on the aerial biomass is less than 5% compared to the chemical fertilizer alone, whereas the combinations according to the invention (frass + pupariums, modalities 9 to 11) show a performance of at least 10.4% compared to frass alone. • At +74 days, these trends are confirmed by the measurement of dry aerial biomass (see Table 2):

[0125] [Tables2] Modality Frass Pupariums Dry aerial biomass 74d vs modality 8 vs modality 10 8 100% 0% 21.2 9 100% 0.50% 23 8.5% 10 90% 0.50% 22.6 11 90% 1% 27.3 28.8% 20.8%

[0126] For methods 2 to 7, the dry aerial biomass at 74 days varies between 27.2g and 28.6g. For method 1 (chemical fertilizer alone), the dry aerial biomass at 74 days is 27.7g.

[0127] For modalities 12 to 18, the dry aerial biomass at 74 days varies between 18.8g and 19.8g.

[0128] The dry aerial biomass of modalities 9 to 11 (11 in particular) is comparable to that of modalities 2 to 7 (with chemical fertilizer instead of frass, and without water stress).

[0129] Thus, these results show that the performance of the puparium + chemical fertilizer combinations (modalities 2 to 7) on dry aerial biomass is less than 5% compared to the chemical fertilizer alone, whereas the combinations according to the invention (frass + pupariums, modalities 9 to 11) show a performance of at least 8.5% compared to frass alone. A significant improvement in plant growth (measured by the amount of root biomass at +54 days and +74 days after sowing) for the combined use according to the invention: • At +54 days (see Table 3): • For the same dose of frass (90%), an increase in the addition of 0.5% of puparium ground material (from 0.5% to 1.0%) improves performance by +29.3%; and • For a lower dose of frass (90% vs. 100%), an addition of 1.0% puparium ground improves performance by +35.8%.

[0130] [Tables3] Modality Frass Puparium s Root biomass 54d vs modality 8 vs modality 10 8 100% 0% 123.6 9 100% 0.50% 120.1 -2.8% 10 90% 0.50% 129.8 11 90% 1% 167.8 35.8% 29.3%

[0131] For modalities 2 to 7, the root biomass at 54 days varies between 134.9g and 157.3g. For modality 1 (chemical fertilizer alone), the root biomass at 54 days is 159.4g.

[0132] For modalities 12 to 18, the root biomass at 54 days varies between 106.6g and 123.3g.

[0133] The root biomass of modalities 9 to 11 is comparable to that of modalities 2 to 7 (with chemical fertilizer instead of frass, and without water stress).

[0134] Thus, these results show that the performance of the puparium + chemical fertilizer combinations (modalities 2 to 7) on the root biomass is less than -0.02% compared to the chemical fertilizer alone, whereas the combinations according to the invention (frass and pupariums, modalities 10 and 11) show a significantly superior performance compared to frass alone. • At +74 days, these trends are confirmed by the measurement of root biomass dry (see Table 4):

[0135] [Tables4] Modality Frass Puparium s Root biomass 74d vs modality 8 vs modality 10 8 100% 0% 12.8 9 100% 0.50% 11.1 -13.3% 10 90% 0.50% 12.9 11 90% 1% 17.5 36.7% 35.7%

[0136] For methods 2 to 7, the dry root biomass at 74 days varies between 13.8g and 15.9g. For method 1 (chemical fertilizer alone), the dry root biomass at 74 days is 16.7g.

[0137] For modalities 12 to 18, the dry root biomass at 74 days varies between 10g and 13.8g.

[0138] The dry root biomass of modalities 9 to 11 is comparable to that of modalities 2 to 7 (with chemical fertilizer instead of frass, and without water stress).

[0139] Thus, these results show that the performance of the puparium + chemical fertilizer combinations (modalities 2 to 7) on the dry root biomass is less than -0.05% compared to the chemical fertilizer alone, whereas the combination according to the invention (90% frass + 1% pupariums, modality 11) shows a performance greater than 35% compared to frass alone.

[0140] These tests do not show any significant impact on plant growth (measured by aerial or root biomass) when the puparium comminuted material is combined with a chemical fertilizer (i.e. no significant difference between methods 1 and 2-7).

[0141] These tests do not show any significant impact on plant growth (measured by aerial or root biomass) when the puparium comminuted material is combined with a chemical fertilizer in a situation of water stress (i.e. no significant difference between methods 12 and 13-18).

[0142] On the other hand, these tests show a significant, even synergistic, impact on plant growth (measured by aerial or root biomass) when the puparium comminuted material is combined with frass.

Claims

Claims

1. Combined use of (A) a first composition comprising (i) holometabolous insect puparia, (ii) whole dead flies or fragments thereof, or (iii) a mixture thereof, and (B) a second composition comprising animal manure, for accelerating plant growth and / or for increasing crop yield, wherein the puparia (i) are Hermetia illucens puparia and have the following dry matter profile: a content of less than or equal to 25% by weight of chitin relative to the total weight of puparia;a content of at least 5% by weight of total fat relative to the total weight of puparium, and manganese in a content of at least 200 ppm on dry matter, and the whole dead flies or their fragments (ii) are Hermetia illucens and have the following profile, by weight of dry matter relative to the total weight of dead flies: a content of less than or equal to 10% by weight of chitin; a content of at least 25% by weight of total fat, and manganese in a content of less than 50 ppm on dry matter.;

2. Use according to claim 1, which is a use of (A) a first composition comprising puparia of Hermetia illucens, and (B) a second composition comprising frass of Hermetia illucens.

3. Use according to one of claims 1 or 2, characterized in that the first and second compositions are two separate compositions, or are previously mixed.

4. Use according to one of the preceding claims, characterized in that the aerial biomass (i.e. leaves, stems and flowers) of the plants is increased by at least 10%, preferably at least 15%, preferably at least 20%, preferably at least 25%, compared to the same species of plant treated with a control, after at least 15 days of culture, preferably at least 30 days of culture, preferably at least 54 days of culture; and / or the root biomass of the plants is increased by at least 15%, preferably at least 20%, preferably at least 30%, preferably at least 35%, compared to the same species of plant treated with a control, after at least 15 days of culture, preferably at least 30 days of culture, preferably at least 54 days of culture, preferably the control is the same animal manure as that present in the combined use, but used alone, preferably the control is Hermetia illucens frass.

5. Use according to one of the preceding claims, characterized in that the first composition is a powder or a paste, preferably obtained by grinding puparia, whole dead flies or their fragments, or their mixtures.

6. Use according to one of the preceding claims, characterized in that the puparia (i) of Hermetia illucens have the following profile, in dry matter: from 5% to 25% by weight, preferably from 10% to 25% by weight, of chitin relative to the total weight of puparium; from 5% to 30% by weight, preferably from 5% to 20% by weight, of total fat relative to the total weight of puparium; and from 200 ppm to 2000 ppm of manganese, preferably from 400 ppm to 1500 ppm, preferably from 600 ppm to 1000 ppm, and / or the whole dead flies or their fragments Hermetia illucens have the following profile, by weight of dry matter relative to the total weight of dead flies: a content of less than or equal to 7% by weight of chitin relative to the total weight of dead flies; from 27% to 40% by weight of total fat relative to the total weight of dead flies; and from 5 ppm to 40 ppm of manganese.

7. Use according to one of the preceding claims, characterized in that the plants are cereals, preferably corn or wheat, or market garden plants, preferably chosen from salads, potatoes, green beans, cucumbers, spinach and carrots.

8. Use according to one of the preceding claims, characterized in that the first and second compositions are provided by mixing with a substrate, and the first composition is used in an amount of at least 0.05%, preferably at least 0.1%, preferably at least 0.5% by weight relative to the weight of dry substrate, preferably in an amount of between 0.05% and 5%, preferably between 0.1% and 3%, preferably between 0.5% and 2% by weight relative to the weight of dry substrate; and / or the second composition is used in an amount of at least less than 0.5 t / ha of substrate, preferably at least 3 t / ha, preferably in an amount between 0.5 t / ha and 10 t / ha of substrate, preferably between 2 t / ha and 10 t / ha of substrate.

9. A method of growing a plant on a substrate, comprising: a. mixtures of the substrate with (A) a first composition comprising (i) puparia of Hermetia illucens, (ii) whole dead flies or their fragments, or (iii) a mixture thereof, and with (B) a second composition comprising animal manure, preferably frass of Hermetia illucens, the mixtures with (A) and (B) being carried out simultaneously with the substrate, or in the order (A) then (B), or in the order (B) then (A); b. preparing a seedbed with the substrate obtained in step a); then c. cultivation of the seed from step b), in which the puparia (i) are puparia of Hermetia illucens and have the following profile, in dry matter: a content of less than or equal to 25% by weight of chitin relative to the total weight of puparium; a content of at least 5% by weight of total fat relative to the total weight of puparium, and manganese at a content of at least 200 ppm on dry matter, and Whole dead flies or their fragments are Hermetia illucens and have the following profile, in dry matter weight relative to the total weight of dead flies: a content of less than or equal to 10% by weight of chitin; a content of at least 25% by weight of total fat, and manganese in a content of less than 50 ppm on dry matter.

10. Method according to claim 9, characterized in that, in step a), compositions (A) and (B) are premixed before mixing with the substrate; or compositions (A), (B) and the substrate are mixed at the same time; or composition (A) is first mixed with the substrate, then the mixture obtained is then mixed with composition (B); or composition (B) is first mixed with the substrate, then the mixture obtained is then mixed with composition (A).