Animal feed composition comprising nanovesicles and method for obtaining same

Abstract

The invention relates to a composition comprising nanovesicles having a core-vesicle structure, the vesicle comprising a first polymer, which is gelatin, representing from 50 to 80% by weight of the total weight of the polymers of the vesicle, a second polymer, which is alginate, representing from 1 to 50% by weight, for example from 20 to 50% by weight, of the total weight of the polymers of the vesicle, and the core comprising an organic core comprising at least one vegetable oil and / or fish oil representing from 5 to 82% by weight of the total weight of the organic core, at least one active substance selected from a natural carotenoid, a natural extract containing at least one natural carotenoid, an essential oil representing from 18 to 95% by weight of the total weight of the organic core. The present invention relates to animal nutrition and more particularly to a composition for supplementing animal feed.

Description

[0001] Composition for animal feed comprising nano-vesicles and method of production

[0002] TECHNICAL FIELD

[0003] The present invention relates to animal nutrition and more particularly to a composition for supplementing animal feed. The invention will find particular application in poultry species and monogastric animal species to improve zootechnical performance, the quality of animal products, and / or animal health. PRIOR TECHNOLOGY

[0004] Animal feed, particularly in agricultural farming conditions, is commonly supplemented with various feed supplements intended to improve health conditions, growth and / or the organoleptic qualities of the finished products.

[0005] Enriched feed, for example with added proteins, lipids, carbohydrates, or supplemented with additives such as vitamins, nutrients, and natural extracts, is widely used. Furthermore, the composition of feed supplements or additives for animals must comply with strict regulations, especially in the context of livestock farming that adheres to specifications dictated by consumer needs.

[0006] Additives and supplements are provided to animals through feed or drinking water. They may undergo physical (extrusion, cooking, etc.) or chemical (saponification / esterification, etc.) processing to modify their bioavailability, targeted release, or stability.

[0007] The traditional approach is to incorporate the active ingredients directly into the food. Generally, additives are added alone or as part of a premix of additives. A primary drawback of incorporating active ingredients directly into the food is their exposure to high temperatures during the various processing stages, as well as to oxidation during and after processing, leading to their degradation.

[0008] Another solution for distributing lipid-soluble active ingredients via drinking water is emulsification. An emulsion is a heterogeneous mixture of two immiscible liquids, one dispersed as fine droplets within the other. Emulsification is achieved by adding surfactants such as soy lecithin. These surfactants stabilize the lipid droplets in aqueous solution. However, these solutions are unstable over the long term.

[0009] Recent solutions allow for the encapsulation of certain additives in micrometric capsules that are then incorporated into food. Various encapsulation techniques are used, such as spray freezing or spray drying. This encapsulation protects the active ingredients from environmental or gastric / intestinal conditions. The benefit is increased stability of the active ingredients and modification of their bioavailability. These microencapsulation techniques make it possible to obtain capsules ranging in size from 1 to 999 µm.

[0010] However, there is always a search for compositions that improve the properties of the finished product, the health status of farm animals, their zootechnical performance and / or the quality of the finished products.

[0011] SUMMARY

[0012] The present invention relates to a composition for animal feed comprising nanometric vesicles of core-shell structure, the shell comprising a first polymer being gelatin representing 50 to 80% by weight of the total weight of the polymers of the shell, a second polymer being alginate representing 1 to 50% by weight of the total weight of the polymers of the shell and the core comprising an organic core comprising at least a vegetable oil and / or a fish oil representing 5% to 82% by weight of the total weight of the organic core, the organic core comprising at least one active ingredient selected from a natural carotenoid, a natural extract containing at least one natural carotenoid and / or an essential oil.

[0013] The invention protects the organic core, advantageously containing fat-soluble active ingredients, from digestive activity and emulsification by bile salts until it exits the duodenum, using a novel encapsulation technique to enhance the efficacy of the active ingredients after this part of the digestive tract. Zootechnical performance is thus remarkably improved. A significant improvement in the quality of livestock products is also observed.

[0014] These results are achieved through the use of nanovesicles (below 1 µm) and the specific characteristics of the shell, leading to targeted release of the organic core in the digestive tract and increased bioavailability after release. The encapsulation technique and the specific characteristics of the resulting shell protect the active ingredients from environmental and gastric conditions and deliver them to a precise point in the digestive tract, ideally after the duodenum. The nanoscale of the nanovesicles increases the contact surface area between the released active ingredients and the intestinal epithelium or gut microbiota. Compared to a micrometric solution, the active ingredients have improved bioavailability and / or a larger contact surface area.

[0015] It has been found surprisingly that the composition according to the invention has an improved feed conversion ratio and that the zootechnical performance of the supplemented animals as well as the quality of the animal products are also greatly improved with this composition.

[0016] According to another aspect, the invention relates to a method for manufacturing the composition as described above, comprising the following steps of

[0017] a. preparation of an organic core solution by mixing at least one vegetable oil and / or fish oil, and an active ingredient in the proportions described above, b. mixing the first polymer and the second polymer in solution,

[0018] c. formation of coacervates by changes in pH or temperature,

[0019] d. addition of the organic nucleus solution obtained in step a. with the coacervates obtained in step c.,

[0020] e. emulsification and / or ultrasonication of the mixture obtained in step d. for the production of nanovesicles.

[0021] According to another aspect, the invention relates to the use of the composition as described above in animal feed to improve the quality of finished products and the zootechnical performance of farm animals.

[0022] According to another aspect, the invention relates to a method for improving finished poultry farming products and zootechnical performance comprising adding the composition as described above to the feed in a quantity of between 10 and 2000 grams of composition per ton of feed intended for poultry farming.

[0023] According to another aspect, the invention relates to a method for improving finished poultry farming products and zootechnical performance comprising adding the composition as described above to drinking water in a quantity of between 0.1 and 5 mL of composition per liter of drinking water intended for poultry farming.

[0024] BRIEF DESCRIPTION OF THE FIGURES

[0025] The aims, objects, features and advantages of the invention will become clearer from the detailed description of an embodiment thereof, which is illustrated by the following accompanying drawings in which:

[0026] Figure 1 represents an observation of a composition according to the invention in transmission electron microscopy treated by a negative staining technique in order to provide contrast.

[0027] Figure 2 represents an observation of a composition according to the invention in transmission electron microscopy treated by a negative staining technique in order to provide contrast.

[0028] DETAILED DESCRIPTION Before beginning a detailed review of embodiments of the invention, optional features that may be used in combination or alternatively are stated below:

[0029] According to one example, at least one natural carotenoid is chosen from among capsanthin, lutein, beta-carotene or bixin.

[0030] For example, at least one natural extract containing at least one natural carotenoid is chosen from paprika extract, marigold extract, cabbage extract, and / or annatto extract. For example, the essential oil is chosen from paprika, rosemary, oregano, garlic, thyme, clove, and / or cinnamon essential oils.

[0031] In one example, the active ingredient represents 18 to 95% by weight of the total weight of the organic core. In another example, the shell comprises a third polymer representing 1 to 50%, preferably 1 to 30%, by weight of the total weight of the shell.

[0032] According to one example, the organic core includes at least one surfactant representing 1 to 2% by weight of the total weight of the organic core.

[0033] According to one example, the surfactant is chosen from soy lecithin, rapeseed lecithin or sunflower lecithin or tween 50 or tween 20.

[0034] For example, the vegetable oil is chosen from rapeseed oil, sunflower oil, linseed oil, palm oil, coconut oil, olive oil.

[0035] For example, fish oil is chosen from cod, salmon, sardine, anchovy, and mackerel oil.

[0036] In one example, the composition is in the form of an aqueous solution.

[0037] In one example, the composition is in a solid form.

[0038] According to one example, the organic core solution is added in a proportion of between 50 and 99% of the total weight of the mixture obtained in step d.

[0039] One example is the use of the composition for animal feed to improve the quality of finished products and the zootechnical performance of farm animals.

[0040] For example, the composition is used for feeding monogastric animals. For example, the composition is used to improve egg quality.

[0041] A parameter "approximately equal to / greater than / less than" or "of the order of" a given value means that this parameter is equal to / greater than / less than the given value, to within 10% or even 5% of that value.

[0042] For the purposes of this disclosure, "A and / or B" means (A), (B), or (A and B). For the purposes of this disclosure, "A, B, and / or C" means (A), (B), (C), (A and B), (A and C), (B and C), or (A, B, and C).

[0043] The terms "first", "second" and "third", "additional", etc. are used simply as labels, and are not intended to impose numerical requirements on their objects.

[0044] By "nanometric" we mean that the smallest dimension of the element is nanometric, that is to say that it is less than or equal to 999nm, preferably 900nm, preferably 800nm, preferably 700nm, preferably 600nm, preferably 500nm, preferably 400nm, preferably 300nm, preferably 200nm, preferably 100nm, preferably 50nm.

[0045] Zootechnical performance refers to the various indicators used to measure the productivity of farm animals. For example, the following parameters can be considered when evaluating zootechnical performance for monogastric animal species: mortality, feed intake, egg production, average egg size, feed conversion ratio, body weight, egg quality (individual egg weight, shell strength, Haugh units, yolk color and yolk weight), meat quality (pH, meat defects (water exudation, color), myodegeneration, white striping (WS), wooden breast (WB), spaghetti muscle, myopathies, muscle hardness and tenderness), nutrient digestibility, egg fertility rate, egg hatching rate, chick quality, litter size per sow, number of live births per sow, number of weaned piglets per sow.

[0046] The composition according to the invention may also be called a complementary feed for animal nutrition, an additive for animal nutrition or a premix of additives for animal nutrition.

[0047] According to one embodiment, the composition is intended to be incorporated into a food intended for animal feed.

[0048] According to another embodiment, the composition is intended to be incorporated into drinking water intended for animal feed.

[0049] According to the invention, the composition comprises nano-vesicles, also known as nanoparticles or nanocapsules. Nano-vesicles are defined as vesicles of nanometric size.

[0050] According to the embodiment, the composition comprises nano-vesicles, of which at least 80%, preferably 90%, have a diameter of less than 1000 nm.

[0051] Advantageously, nanovesicles have a core-shell structure comprising a shell made of polymers enclosing an organic core.

[0052] According to the invention, the shell comprises at least two polymers and can, according to embodiments, comprise three or more polymers.

[0053] According to the invention, the nano-vesicle, and more particularly the shell, comprises a first polymer. Preferably, the first polymer is fish gelatin, bovine gelatin, or porcine gelatin.

[0054] According to the invention, the nano-vesicle, and more particularly the shell, comprises a second polymer. Preferably, the second polymer is an alginate.

[0055] One possibility is that the nano-vesicle, and more specifically the shell, includes a third polymer. For example, the third polymer is chosen from among gum arabic and / or xanthan gum and / or agar and / or pectin and / or cellulose and / or carboxymethylcellulose.

[0056] According to the invention, the nano-vesicle, and more preferably the shell, comprises from 50 to 80% by weight of the first polymer relative to the total weight of the polymers in the shell. According to the invention, the nano-vesicle, and more preferably the shell, comprises from 1 to 50%, preferably from 20 to 50% by weight of the second polymer relative to the total weight of the polymers in the shell.

[0057] According to the invention, the nano-vesicle, and more preferably the shell, comprises from 1 to 50%, preferably from 1 to 30% by weight of the third polymer by weight relative to the total weight of the polymers of the shell.

[0058] According to the invention, the nano-vesicle, more preferably the core, comprises an organic core. The organic core comprises at least one compound having an active effect in animal nutrition. The organic core comprises the active ingredient(s) intended to be protected by the encapsulation and the presence of the vesicle.

[0059] The organic core comprises at least one vegetable oil or a mixture of vegetable oils and / or one fish oil or a mixture of fish oils. Vegetable oil is defined as a fat derived from oilseeds or oilseeds. Fish oil is an oil obtained from the biological tissues of oily fish.

[0060] The vegetable oil or mixture of vegetable oils is chosen from rapeseed oil, sunflower oil, linseed oil, palm oil, coconut oil, olive oil.

[0061] Fish oil or a mixture of fish oils is chosen from cod, salmon, sardine, anchovy, and mackerel oil.

[0062] Advantageously, at least one vegetable oil and / or fish oil is present in the organic core in a quantity between 30% and 80% by weight of the total weight of the organic core.

[0063] The organic core of the invention comprises, according to the invention, at least one active ingredient selected from a natural carotenoid, a natural extract containing at least one natural carotenoid, and an essential oil. The present invention surprisingly improves the zootechnical performance of animals whose diet is supplemented by the composition, while ensuring advantageous release after the duodenum, i.e., after the site of absorption of the carotenoids and essential oils. This is evidenced, for example, by the minimal effect of the composition on the pigmentation of the finished products. The nano-vesicle according to the invention creates a protective barrier around the organic core for targeted release after the duodenum, thus surprisingly improving the zootechnical performance of the animals.

[0064] The term carotenoid encompasses molecules from the carotene and xanthophyll families, also known as tetraterpenes. Carotenoids are fat-soluble pigments found in a wide variety of living organisms.

[0065] The term natural carotenoid refers to a carotenoid obtained from nature and differs from synthetic carotenoids. For example, natural carotenoids include capsanthin, lutein, and beta-carotene, while synthetic carotenoids include apo-esters and canthaxanthin.

[0066] Preferably, the natural extract containing at least one carotenoid is chosen from marigold (Tagetes erecta) extract, paprika (Capsicum annuum) extract, annatto (Bixa orellana) extract, and cabbage (Brassica oleracea) extract. Specifically, paprika extract is paprika oleoresin, also known as E160 c(i). Paprika oleoresin is extracted from the fruits of the species Capsicum annuum and / or Capsicum frutescens (chili peppers).

[0067] Preferably, marigold extract, annatto extract and common cabbage extract are also fat-soluble extracts.

[0068] The organic core comprises, in one embodiment, at least one essential oil. An essential oil is defined as an oil extracted from aromatic plants by steam distillation of various plant parts (bark, leaves, flowers, etc.) or by cold pressing of fresh citrus peels. For example, the essential oil included in the organic core advantageously comprises molecules with antioxidant, anti-inflammatory, bacteriostatic, and / or antiparasitic activity. For example, the organic core comprises an essential oil or a mixture of essential oils selected from essential oils containing rosemarinic acid, carnosol, thymol, eugenol, limonene, menthol, carnosic acid, cinnamaldehyde, carvacrol, trans-anethole, and / or allicin.As an example, the essential oil or a mixture of essential oils is chosen from paprika, rosemary, oregano, savory, garlic, thyme, lemon, peppermint, star anise, clove and / or cinnamon essential oil.

[0069] Advantageously, at least one active ingredient selected from at least one natural carotenoid, a natural extract containing at least one natural carotenoid, or an essential oil is present in the organic core in a quantity of between 18% and 95% by weight of the total weight of the organic core. At least one vegetable oil and / or at least one fish oil, possibly with a surfactant, represents between 5% and 82% by weight of the total weight of the organic core.

[0070] According to an advantageous embodiment, the organic core comprises at least one surfactant. The surfactant is, for example, selected from soy lecithin, rapeseed lecithin, or sunflower lecithin, or from hydrolyzed versions of these same lecithins, or from tween 50 or tween 20.

[0071] Advantageously, the surfactant is present in the organic core in an amount between 0.5% and 3% by weight of the total weight of the organic core.

[0072] According to one embodiment, the composition comprises at least one additive intended for the stability of the composition. For example, carrageenan, gum arabic, guar gum or xanthan gum, starch, vegetable fibers or a mixture of these ingredients.

[0073] The encapsulation capacity of the nano-vesicles according to the invention is between 90 and 100%. Encapsulation capacity is understood as the percentage of the material intended to be encapsulated that is actually encapsulated.

[0074] The proportion of organic nucleus in a nanovesicle is between 50 and 99% by weight of the total weight of the nanovesicle.

[0075] In one possibility, the composition is in liquid form. Advantageously, the composition is an aqueous solution comprising nanovesicles. In another possibility, the composition is in a solid state, having undergone atomization treatment of an aqueous solution.

[0076] Surprisingly, the applicant identified that the use of the composition in animal feed improves the quality of finished products from monogastric farm animals as well as zootechnical performance.

[0077] The term "finished products" refers to monogastric livestock products, including eggs, milk, colostrum, muscle, gametes, and organs. Monogastric livestock are preferably understood to mean laying hens, broiler chickens, and pigs. To this end, the invention relates to the use of the composition by addition to animal feed and / or drinking water intended for animal feed.

[0078] According to a first embodiment, the composition is added in a quantity of between 10 and 2000 grams of composition per tonne of feed intended for animal nutrition, preferably in a quantity of between 500 and 1000 grams per tonne.

[0079] According to a second embodiment, the composition is added in a quantity of between 0.1 and 5 mL of composition per litre of drinking water intended for animal feed, preferably in a quantity of between 0.2 and 2 mL of composition per litre of drinking water intended for animal feed.

[0080] According to one aspect, the invention relates to a method for preparing the composition as described above.

[0081] The process advantageously includes encapsulation of the organic core by the selected polymer(s) followed preferably by a step of reducing the size of the vesicles to obtain nano-vesicles of nanometric size.

[0082] According to a preferred embodiment, the encapsulation of the organic nucleus is achieved by coacervation.

[0083] The principle of coacervation is based on the attraction between polymers with complementary charges to form vesicles.

[0084] The process includes a step a. of preparing the organic core. This step a. of preparing the organic core comprises mixing at least one active ingredient selected from a natural carotenoid, a natural extract containing a natural carotenoid, an essential oil in at least one vegetable oil and / or at least one fish oil, according to the proportions described above. An essential oil is advantageously added. Advantageously, a surfactant is also added. Depending on the option, additives, for example, for stability as detailed above, suitable for animal feed, may also be added. The mixing is carried out under mechanical agitation with a static mixer for 10 minutes. The process of preparing the composition includes a step b. of mixing the selected polymers. In particular, the first polymer, the second polymer, and optionally the third polymer are mixed in solution.The solution is an aqueous solution. The mixture is prepared under mechanical agitation with a static mixer for 10 minutes.

[0085] The composition preparation process includes a step c, coacervate formation. This step involves the formation of bonds between the polymers, specifically the first, second, and third polymers. This step c is performed on the polymer mixture in solution obtained in step b. This step involves a change in pH and / or temperature.

[0086] According to one possibility, step c. includes the addition of an acid such as citric, formic, propionic, lactic, benzoic acid, or a mixture of these acids to achieve a pH below 4.5.

[0087] According to another possibility, step c. involves a temperature change of the polymer solution. For example, the polymer solution is heated to a temperature of 30 to 50°C. Preferably, the heating is carried out by means known to those skilled in the art. Advantageously, the heating has a maximum duration of 15 minutes. For example, stirring is carried out simultaneously with the heating of the aqueous solution in a tank.

[0088] At the end of step c, corresponding to a coacervation step, coacervates and vesicles are formed.

[0089] The process includes a step d. of adding the organic nucleus solution, obtained in step a., with the coacervates obtained in step c.

[0090] The organic core solution is then integrated into coacervates forming vesicles having the organic core and a polymer shell at their core.

[0091] According to the invention, the process includes an emulsification step e. This step e. is performed on the mixture obtained at the end of step d. to form a solution comprising the nanovesicles. Advantageously, the emulsification step is carried out before the vesicles have completely solidified. The emulsification is advantageously achieved by a continuous process comprising high-shear mixing and / or high-pressure homogenization. This step is advantageously carried out within one hour of step d.

[0092] Emulsification makes it possible to reduce the size of vesicles to approach, or even reach, nanometric sizes.

[0093] The process includes an ultrasonication step implemented after the emulsification step. Ultrasonication reduces the size of the vesicles to nanometer sizes.

[0094] Ultrasonication involves applying ultrasonic waves to the emulsion obtained at the end of step e in a continuous, i.e., direct, process. The preparation is exposed to ultrasound for less than one minute with a power range of 0.5 to 5 W / mL / min. According to the advantageous embodiment, the organic core is added in a proportion of 50 to 99% by weight of the final mixture obtained at the end of step d.

[0095] According to one aspect, the invention relates to the use of the composition to improve the laying rate of laying hens.

[0096] According to another aspect, the invention relates to the use of the composition to improve the vitelline structure of laying hen eggs.

[0097] According to another aspect, the invention relates to the use of the composition to improve the maintenance of the protein structure of laying hen egg white over time. According to our aspect, the invention relates to the use of the composition to improve the zootechnical performance of laying hens.

[0098] Examples

[0099] Example 1 Composition

[0100] Aqueous solution comprising nanovesicles comprising

[0101] The first polymer being gelatin, representing 70% by weight of the total weight of the shell's polymers,

[0102] a second polymer being alginate, representing 30% by weight of the total weight of the shell's polymers,

[0103] a core comprising an organic core including at least one vegetable and / or fish oil representing 59% of the total weight of the organic core, at least one active ingredient selected from a natural carotenoid, a natural extract containing at least one natural carotenoid, advantageously extracted from paprika, an essential oil representing 40% of the total weight of the organic core, and at least one surfactant.

[0104] Electron microscopy views are shown in Figure 1 and Figure 2 clearly illustrating the presence of nanovesicles, as well as the homogeneity of nanovesicle size.

[0105] Example 2 trials

[0106] The experiment is conducted on 4 groups of 200 laying hens (divided into 10 replicates of 20 laying hens) receiving different treatments:

[0107] T1 - Control, receiving standard feed and drinking water without supplementation, T2 - Control + organic core according to example 1 in the feed (500 g / T),

[0108] T3 - Control + organic core according to example 1 in the diet (1000 g / T),

[0109] T4 - Control + composition including nano-vesicles containing the organic core according to example 1 in drinking water (1.5 mL / L).

[0110] Treatments T2 and T4 are equivalent in terms of organic core content. Treatment T3 provides twice the dose of organic core content compared to treatments T2 and T4.

[0111] The trial begins at 49 weeks of age. The effects are studied between 52 and 55 weeks of age (duration = 28 days).

[0112] The experiment is divided into 2 periods:

[0113] The first period (from 49 to 50 weeks of age, duration = 14 days) is placed before the start of the experiment. The objective of the comparison before the start of the experiment is to ensure the comparability of the initial groups.

[0114] Between the 1st and 2nd periods (duration = 7 days): Analysis of the data from the 1st period. The 2nd period (from 52 to 55 weeks, duration = 28 days) is the experimental phase.

[0115] Zootechnical criteria (mortality, feed intake, and egg production) are recorded between 49 and 55 weeks of age. Average egg size and feed intake are assessed weekly. Body weight is estimated at two different ages (49 and 55 weeks). Egg quality characteristics (individual egg weight, shell strength, Haugh units, yolk color, and yolk weight) are measured on one-day-old eggs at 13 different time points.

[0116] 50 weeks old,

[0117] 3 times at 52 weeks of age,

[0118] 3 times at 53 weeks of age,

[0119] 3 times at 54 weeks of age,

[0120] 3 times at 55 weeks of age.

[0121] Mortality registration

[0122] Dead birds are counted between 49 and 55 weeks of age. Mortality is recorded daily by repetition.

[0123] Food consumption recording

[0124] Feed consumption is determined by repeating the measurements weekly during the experimental period (from 49 to 55 weeks of age). The daily feed consumption per bird can then be calculated.

[0125] Egg production

[0126] 5 days a week, the number of "saleable" eggs is recorded by repetition (soft-shelled and very fragile eggs are not recorded).

[0127] Estimating egg size

[0128] Each week, the average size of the eggs in each group is estimated by weighing all the eggs from a day of production.

[0129] Body weight estimation

[0130] Individual body weights are measured at 49 and 55 weeks of age.

[0131] Egg quality measurements

[0132] The quality of the eggs (50 eggs / treatment / age 5 eggs / replicate) is precisely estimated per treatment at 13 different times:

[0133] Once at 50 weeks of age,

[0134] 3 times at 52 weeks of age,

[0135] 3 times at 53 weeks of age,

[0136] 3 times at 54 weeks of age,

[0137] 3 times at 55 weeks of age.

[0138] 10 eggs per treatment are collected the week following the end of the trial.

[0139] Weight of an individual egg

[0140] At each repetition, the eggs are weighed individually using a Sartorius balance (accuracy of 0.1 g).

[0141] Shell strength

[0142] The shell's strength is evaluated using an MTS Synergie 200 compression machine. The compression speed is 30 mm / min. Two different characteristics define the shell's strength: Shell rigidity, which is measured by the shell's deformation under a constant force of 15 Newtons. The less the shell deforms, the stronger it is.

[0143] Breaking force (accuracy of 0.1 Newton) which is the maximum force applied to the egg before the shell breaks.

[0144] Haugh Units

[0145] The albumen height is measured using TSS equipment. Knowing the egg weight (EW) and the albumen height (AH), we can calculate the Haugh units (HU): HU = 100 log (AH - 1.7 EW0.37 + 7.57)

[0146] The color of yellow

[0147] The "Roche yolk color fan" egg yolk colouring scale is used.

[0148] The color of yellow is also measured using a Minolta CR400 chromameter.

[0149] Weight of the yolk

[0150] For each egg, the yolk is weighed using a Sartorius balance (accuracy of 0.1 g).

[0151] Blood samples

[0152] Blood samples are taken at the end of the trial (55 weeks of age):

[0153] - 1 animal per replica,

[0154] - Blood sampling for plasma collection = heparinized tubes,

[0155] - These tubes collected for plasma will be centrifuged in the laboratory and the samples will be kept in a freezer (-18°C).

[0156] Results :

[0157] Criteria for estimating the effect of the experimental product:

[0158] Mortality: Cumulative mortality rates are compared during the experimental period. Dietary intake: Daily dietary intakes are compared during the experimental period.

[0159] Egg production: The weekly production of marketable eggs per replicate is estimated between 49 and 55 weeks of age. Egg production is recorded 5 days per week, so the weekly production is estimated using a simple proportion. The cumulative number of eggs can then be compared over the experimental period.

[0160] Egg mass: The average egg weight is recorded weekly so that we can estimate the cumulative egg mass per replicate. Treatments can then be compared for this trait.

[0161] Feed conversion ratio (FCR) is the ratio of the mass of feed consumed to the mass of eggs. Treatments can also be compared for this trait. A decrease in the index demonstrates improved feed efficiency.

[0162] Body weight: Average body weights are compared at 2 different ages (49 and 55 weeks of age).

[0163] Results for the first period: no difference between the 4 groups.

[0164] Results for the second period:

[0165] Mortality: no significant difference was observed. Laying rate: a significant difference was observed between groups T3 and T4 and the control group T1 and T2. Groups T3 and T4 were higher than T1 and T2.

[0166] Daily food intake: a significant difference was observed between group T2 and the control group T1. Groups T3 and T4 were higher than group T2, while groups T3 and T4 were not different from the control group T1.

[0167] Dietary conversion index: a significant difference was observed between groups T3 and T4 and the control group T1. Group T2 was not significantly different from the control group T1. Group T3 was not significantly different from T4.

[0168] Proportion of yellow: a significant difference is observed between the T3 and T4 groups and the control group T1. The T3 and T4 groups are superior to the T2 group, which is itself superior to the T1 group.

[0169] Yellow color: a significant difference is observed between groups T2 and T3 and the control group T1. Group T3 is superior to group T2, which is itself superior to group T4. There are no significant differences between group T4 and the control group T1. This clearly demonstrates the absence of a pigmenting effect from the natural carotenoid when it is contained within the organic core of the nano-vesicle according to the invention, and the effectiveness of protecting the organic core from gastric and intestinal conditions.

[0170] Albumen height during the trial: a significant difference was observed between the T4 group and the control group T1. The T4 group was superior to the T1 group, which was itself equivalent to the T3 group, which were themselves significantly superior to the T2 group.

[0171] Albumen height analyzed 5 weeks after laying during the trial: a significant difference is observed between the T4 group and the control group T1. This demonstrates a significant improvement in egg white quality over a storage time of more than 5 weeks.

[0172] These results demonstrate an improvement in zootechnical performance in groups T4 and T3 compared to the control group T1 and even compared to group T2. Group T4 showed the greatest improvement in zootechnical performance and egg quality parameters relative to the amount of the composition administered to the animals, and in particular the amount of carotenoids. Indeed, using a lower quantity of carotenoids, resulting in cost savings, is advantageous for achieving a superior or even equivalent effect. Thus, the composition according to the invention used in group T4, which, thanks to nanovesicles, protects the organic nucleus and allows its release after the duodenum, is particularly advantageous compared to using a single organic nucleus without the protection technology of the present invention.

Claims

Demands 1. Composition intended for animal feed comprising nanometric vesicles of core-shell structure, the shell comprising a first polymer being gelatin representing 50 to 80% by weight of the total weight of the polymers of the shell, a second polymer being alginate representing 1 to 50% by weight of the total weight of the polymers of the shell and the core comprising an organic core comprising at least a vegetable oil and / or a fish oil representing 5% to 82% by weight of the total weight of the organic core, the organic core comprising at least one active ingredient selected from a natural carotenoid, a natural extract containing at least one natural carotenoid and / or an essential oil.

2. Composition according to the preceding claim wherein at least one natural carotenoid is selected from capsanthin, lutein, beta-carotene or bixin.

3. A composition according to any one of the preceding claims, wherein at least one natural extract containing at least one natural carotenoid is selected from paprika extract, marigold extract, cabbage extract, and / or annatto extract.

4. A composition according to any one of the preceding claims, wherein the essential oil is selected from paprika, rosemary, oregano, garlic, thyme, clove, and / or cinnamon essential oils.

5. Composition according to any one of the preceding claims wherein the active ingredient represents from 18 to 95% by weight of the total weight of the organic core.

6. Composition according to any one of the preceding claims wherein the shell comprises a third polymer representing from 1 to 50% by weight of the total weight of the shell.

7. Composition according to any one of the preceding claims wherein the organic core comprises at least one surfactant representing 1 to 2% by weight of the total weight of the organic core.

8. Composition according to the preceding claim wherein the surfactant is selected from soy lecithin, rapeseed lecithin or sunflower lecithin or tween 50 or tween 20.

9. Composition according to any one of the preceding claims wherein the vegetable oil is selected from rapeseed oil, sunflower oil, linseed oil, palm oil, coconut oil, olive oil.

10. Composition according to any one of the preceding claims wherein the fish oil is selected from cod, salmon, sardine, anchovy, mackerel oil.

11. Composition according to any one of the preceding claims being in the form of an aqueous solution.

12. Composition according to any one of claims 1 to 10 being in solid form.

13. Composition according to any one of claims 1 to 12 wherein the alginate represents from 1 to 50% by weight of the total weight of the shell polymers.

14. A method for manufacturing the composition according to any one of the preceding claims, comprising the following steps of a. preparation of an organic core solution by mixing at least one vegetable oil and / or fish oil, and an active ingredient in the proportions according to claim 1, b. mixing the first polymer and the second polymer in solution, c. formation of coacervates by changing the pH or temperature, d. addition of the organic core solution obtained in step a. with the coacervates obtained in step c. e. emulsification and ultrasonication of the mixture obtained in step d. to obtain nano-vesicles.

15. Manufacturing process according to the preceding claim in which the organic core solution is added in a proportion of between 50 and 99% of the total weight of the mixture obtained in step d.

16. Use of the composition according to any one of claims 1 to 13 in animal feed to improve the quality of finished products and the zootechnical performance of farm animals.

17. Use according to the preceding claim for feeding monogastric animals.

18. Use according to claim 16 to improve egg quality.

19. A method for improving finished poultry farming products and zootechnical performance comprising adding the composition according to any one of claims 1 to 13 to the feed in a quantity of between 10 and 2000 grams of composition per tonne of feed intended for poultry farming.

20. A method for improving finished poultry farming products and zootechnical performance comprising adding the composition according to any one of claims 1 to 13 to drinking water in an amount of between 0.1 and 5 mL of composition per liter of drinking water intended for poultry farming.

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