FISH PROTEIN HYDROLYSATE AND APPLICATIONS IN FARM ANIMALS

Abstract

FISH PROTEIN HYDROLYSATE AND APPLICATIONS IN FARM ANIMALS The present invention relates to a protein hydrolysate originating from fish tissues or cartilages rich in protein, in particular collagen, and its applications in the feeding and welfare of farm animals, in particular pigs and poultry. In particular, the invention relates to a hydrolysate obtained by enzymatic digestion using an endopeptidase enzyme, said hydrolysate being characterized in that it has a protein fraction having a molecular profile with the following distribution: between 15 and 60% of peptides with a molecular weight of between 1000 and 5000 Da, between 20 and 40% of peptides with a molecular weight of between 500 and 1000 Da, between 10 and 40% of peptides with a molecular weight of between 150 and 500 Da, between 1 and 10% of peptides with a molecular weight of between 1000 and 50 ... molecular weight less than 150 Da. Fig. 2

Description

Title of the invention: FISH PROTEIN HYDROLYSATE AND APPLICATIONS IN FARM ANIMALS Technical field

[0001] The present invention is in the field of animal welfare and relates to a protein hydrolysate originating from fish tissues or cartilages rich in proteins, in particular collagen, as well as a process for its production. The hydrolysate according to the invention finds application in feeding and maintaining good physical condition of farm animals, in particular pigs and poultry. In particular, the hydrolysate has anti-inflammatory and antioxidant effects limiting or inhibiting pain phenomena and loss of physical functions, prevents or treats bursitis phenomena and allows the animal to be kept in conditions promoting or stimulating the animal's growth. STATE OF THE PRIOR ART

[0002] The pig growth cycle is generally divided into several stages, from birth to slaughter.

[0003] Piglets are born after a gestation period of about 114 days. Piglets weigh between 1.2 and 1.5 kg at birth. They stay with their mother for about three to four weeks, and are nursed.

[0004] The next stage is the so-called post-weaning phase, which lasts approximately six to eight weeks. The piglets are separated from their mothers and brought together in suitable pens; this is called weaning. They then move from a milk diet to a solid plant-based diet.

[0005] This is followed by the fattening stage lasting three to four months. At the end of this period, the pig, called a charcutier pig, is sent to the slaughterhouse at a live weight of around 120 kg.

[0006] During the post-weaning and fattening stages, growth problems can appear, thus harming the animal's performance. Piglets are in fact subjected to different sources of stress, such as dietary stress linked to the change of diet, environmental stress linked to the change of housing and social stress linked to their separation from their mother. Oxidative stress reactions and inflammatory reactions are thus observed during this period, often linked to oxidative stress.

[0007] Such a condition is detrimental to the animal's growth, i.e. its weight gain. The same phenomena are also observed in other livestock, such as poultry.

[0008] Another factor that affects the growth of piglets is the occurrence of bursitis. Bursitis is caused by an inflammatory reaction of the bursa, a small sac located near the joints that contains a small amount of fluid that facilitates the movement of tendons and muscles on adjacent bones. When this bursa becomes inflamed, it is called bursitis.

[0009] Bursitis can be caused by injuries, trauma, or irritation. Conditions that promote oxidative stress and inflammation are also likely aggravating factors. In pigs, it usually appears on the lower part of the hind limbs, at the tarsals and metatarsals. Bursitis is very visible, causing swelling, tenderness, redness, and possibly lameness. Treatment depends on the underlying cause of the bursitis and may include measures such as the administration of anti-inflammatory drugs.

[0010] The solutions currently proposed to improve the welfare of farm animals, such as pigs and poultry, are turning to natural approaches. For example, the intake of vitamin E is proposed to combat oxidative stress. Several polyphenols, such as resveratrol, catechins, quercetin or curcumin, are also considered to reduce oxidative stress. However, they do not appear to be effective in treating inflammatory reactions. Regarding bursitis in pigs, the treatments currently proposed are based solely on taking anti-inflammatory drugs, applying antiseptic creams or improving the quality of the floors in pens.

[0011] As is often the case, the most satisfactory approach is prevention. It must aim to restore normal growth and animal well-being. Controlling risk factors in livestock farming, particularly soil quality, is an essential but unfortunately often insufficient prerequisite. In addition, action on the mechanisms responsible for inflammation and oxidative stress is desirable in order to achieve the above objective. Statement of the invention

[0012] The aim of the present invention is thus to propose a natural and effective solution for the well-being of farm animals, such as pigs and poultry, in particular to prevent and treat oxidative stress and inflammation reactions and to limit or treat bursitis phenomena, and to promote weight gain and growth of the animal.

[0013] To this end, the invention relates to a hydrolyzate of tissue or cartilage proteins rich in fish collagen, obtained by enzymatic digestion using an endopeptidase enzyme of bacterial origin, said hydrolyzate being characterized in that that it presents a protein fraction with a molecular profile of the following distribution: - between 15 and 60% of peptides with a molecular weight between 1000 and 5000 Da, - between 20 and 40% of peptides with a molecular weight between 500 and 1000 Da, - between 10 and 40% of peptides with a molecular weight between 150 and 500 Da, - between 1 and 10% of peptides with a molecular weight of less than 150 Da,

[0014] and an aminogram in which, as a percentage of the total nitrogenous matter: - the glutamic acid content is between 10 and 14% - the valine content is between 1.5 and 3.5% - the isoleucine content is between 1 and 3% - leucine content is between 2 and 6% - the glycine content is between 9 and 13% - the lysine content is between 0.5 and 3% - the arginine content is between 6 and 10%.

[0015] The hydrolyzate according to the invention is a mixture of peptides and free amino acids obtained after enzymatic hydrolysis, using a specific enzyme, of raw materials originating from fish, such as fish skins and cartilages. These materials being rich in collagen, the peptides and free amino acids composing the hydrolyzate are mainly derived from collagen.

[0016] The hydrolyzate according to the invention proves to be particularly effective in stimulating the growth of livestock, as shown by the following examples.

[0017] It will also be noted that the hydrolyzate according to the invention is advantageously rich in amino acids precursors of growth hormone such as arginine, valine, glutamic acid, lysine and long-chain amino acids BCAA, i.e. leucine, isoleucine and valine. Growth hormone increases muscle volume, increases bone density and reduces fat synthesis.

[0018] Furthermore, the hydrolysate according to the invention is effective in preventing and treating bursitis phenomena in pigs, preventing and treating oxidative stress and inflammation reactions, responsible for growth problems in farm animals.

[0019] The antioxidant effect fights against cellular aging.

[0020] The anti-inflammatory effect protects against inflammation, such as redness, swelling, pain and loss of normal physical functions.

[0021] Consequently, the animal treated with a hydrolysate according to the invention is a healthy animal and can therefore continue normal growth.

[0022] Advantageously, the hydrolyzate according to the invention also has:

[0023] - a fat content of less than 0.5% as a percentage of dry extract,

[0024] - a total nitrogenous matter content (N * 6.25) greater than 90%, in percentage dry extract,

[0025] - a mineral content of less than 10%, as a percentage of dry extract.

[0026] Preferably, the aminogram also presents, as a percentage of the total nitrogenous matter:

[0027] - an aspartic acid content of between 7 and 10%

[0028] - a proline content is between 7 and 11%

[0029] - a threonine content is between 2 and 5%

[0030] - a serine content is between 3 and 7%

[0031] - an alanine content is between 8 and 12%

[0032] - a cystine content is between 0.1 and 1%

[0033] - a methionine content is between 1.5 and 3%

[0034] - a tyrosine content is between 0.5 and 2.5%

[0035] - a phenylalanine content is between 1 and 3.5%

[0036] - a histidine content is between 3.5 and 7%

[0037] - a hydroxyproline content is between 4 and 8%

[0038] - a hydroxylysine content is between 0.5 and 2.5%

[0039] Preferably, the tissue rich in fish collagen is skin.

[0040] Preferably again the fish is a Salmonidae, preferably salmon or trout, or even a white fish.

[0041] According to a characteristic of the invention, the enzyme is an endoprotease of bacterial origin marketed under the brand name Corolase 7089 by the company AB enzymes.

[0042] The invention also relates to a process for obtaining a fish protein hydrolysate as described above, the process being characterized in that it comprises the steps of:

[0043] - preparation of a protein source from fish tissues and / or cartilages rich in collagen,

[0044] - preparation of an enzymatic solution comprising between 0.1 and 0.5% of a endoprotease enzyme,

[0045] - enzymatic hydrolysis of said protein source at a temperature between between 40 and 60°C, for 1 to 5 hours, after adding the enzyme solution,

[0046] - stopping the enzymatic hydrolysis by heating to a temperature of at least 90°C for 8 to 20 minutes,

[0047] - separation of the protein hydrolyzate from the other components,

[0048] - recovery and concentration of protein hydrolyzate.

[0049] According to the method of the invention, the enzyme is an endoprotease of bacterial origin marketed under the brand name Corolase 7089 marketed by the company AB enzymes. Advantageously, the protein source / water ratio is less than or equal to 1.

[0050] Preferably, the tissue is skin.

[0051] Preferably, the fish is a Salmonidae, preferably salmon or trout, or even a white fish.

[0052] The invention also relates to a composition comprising a protein hydrolyzate as defined previously, or obtained by a process as defined previously.

[0053] A composition according to the invention may be in any known form suitable for feeding livestock, such as a solution, a solid composition such as a tablet, a powder or a capsule, a granule or even a gel. The composition may comprise any suitable excipient, and any other compound such as vitamins, minerals, active ingredients, preservatives, colorants.

[0054] The invention also relates to a hydrolyzate of proteins from tissues or cartilages rich in fish collagen as defined above, or obtained by a process as defined above, or a composition containing it, for its use in feeding farm animals, in particular pigs and poultry.

[0055] The invention also relates to a hydrolyzate of tissue or cartilage proteins rich in fish collagen as defined above, or obtained by a process as defined above, or a composition containing it, for stimulating the growth of farm animals, in particular pigs and poultry.

[0056] Advantageously, the growth stimulation includes weight gain, in particular muscle gain.

[0057] The invention also relates to a hydrolyzate of tissue or cartilage proteins rich in fish collagen as defined above, or obtained by a process as defined above, or a composition containing it, for the prevention and treatment of bursitis.

[0058] The invention also relates to a hydrolyzate of tissue or cartilage proteins rich in fish collagen as defined above, or obtained by a process as defined above, or a composition containing it, for the prevention and treatment of antioxidant and anti-inflammatory reactions in farm animals, in particular pigs and poultry.

[0059] In the context of the invention, the animals may be healthy animals or animals suffering from growth retardation. Indeed, the treatment of animals using a hydrolyzate according to the invention may be preventive or curative.

[0060] Advantageously, the hydrolyzate, or the composition containing it, is administered to porcine animals during the fattening period, preferably from the 6th week after birth, for 6 weeks, or during the last three to six weeks of the fattening period.

[0061] Preferably again, within the framework of the invention, the hydrolyzate, or the composition containing it, are administered at a rate of 3 to 7 g / day of hydrolyzate. This daily dosage is particularly suitable for a product whose dry extract is 50%.

[0062] Starting from the necessary daily dosage, a person skilled in the art will know how to formulate a composition adapted to such a dosage.

[0063] The above-mentioned features of the invention, as well as others, will appear more clearly on reading the following examples and in connection with the following attached drawing:

[0064] [Fig-1] is a diagram representing the distribution of the molecular weight of protein hydrolysates according to the invention,

[0065] [Fig.2] is a diagram representing the average production of interleukin 6, in pg / pg of protein, after exposure of human keratinocyte cells to UV-B, the cells having been cultured with a hydrolyzate according to the invention at different concentrations, or with vitamin D3, ethanol, or simply a culture medium,

[0066] [Fig.3] is a curve diagram representing the Trolox equivalents of a hydrolysate according to the invention as a function of the Net-AUC (Area under the net curve).

[0067] EXAMPLES

[0068] Example 1: obtaining hydrolysates of salmon skin proteins

[0069] Four hydrolysates of salmon skin proteins were prepared according to the following method: - Mixture of salmon skins with 1 volume of water and 0.2% of Corolase 7089® enzyme - Enzymatic hydrolysis for 1 to 5 hours at a temperature of 40 to 60°C, - Inactivation of the enzyme by heating to a temperature of at least 90°C for 8 to 20 minutes, - Sieving on a 300 pm sieve to eliminate hydrolysis residues, - Centrifugation of the hydrolysis juice for 10 to 15 minutes at 4700 rpm to separate the oil, insolubles and protein hydrolysate, - Concentration of the hydrolyzate.

[0070] Table 1 shows the characteristics of the hydrolysates.

[0071] [T AB. 1] Parameters Composition dry extract Dry extract (%) 100% Total proteins (N*6.25) >90% Fat <0.5% Minerals <10%

[0072]

[0073] [Fig.l] shows the molecular weight distribution of the four hydrolysates. Example 2: evaluation of the anti-inflammatory properties of a hydrolysate according to the invention

[0074] The aim of this test is to evaluate the anti-inflammatory activity of a hydrolysate obtained by a method such as that described in Example 1 on human keratinocyte cells irradiated by exposure to UV-B.

[0075] In this example, human keratinocyte cells are pre-incubated with the test compounds for 24 hours before exposure to UV-B. The supernatants are collected 24 hours after UV-B treatment and the secretion of interleukins 6 (IL-6) is quantified according to the ELISA test.

[0076] Interleukin 6 acts as a messenger between cells involved in the regulation of acute and chronic inflammation. Overproduction of interleukin 6 causes inflammation and may be linked to the origin of joint damage.

[0077] Witness controls:

[0078] Positive control: 1,25-dihydroxy vitamin D3 (0.24 μM diluted in BSA culture medium) is used as a known inhibitor of IL-6 secretion (From HAES et al. (J. Cell. Biochem., 89(4):663-73, 2003)).

[0079] Solvent control: ethanol (0.1% diluted in BSA culture medium) and culture medium.

[0080] Keratinocyte cells: HEK001 cell line

[0081] Protocol:

[0082] At D0, the keratinocyte cells are distributed, in their culture medium, in 12-well plates. They are incubated at 37°C, 5% CO2, to obtain a confluence of less than 50% at DI.

[0083] At Dl, the cells are treated with the following compounds: - Hydrolyzate according to the invention in five concentrations: 0.06 mg / ml; 0.31 mg / ml; 1.57 mg / ml; 7.84 mg / ml; 39.20 mg / ml - 0.24 pM of 1,25-dihydroxyvitamin D3 - 0.1% ethanol - Culture medium

[0084] Each treatment is tested with and without UV-B exposure.

[0085] UV-B treatment:

[0086] The cells are irradiated at a dose of 12 mJ / cm2 of UVB at 312 nm.

[0087] After irradiation, the cells are incubated in fresh culture medium for 24 hours at 37°C-5% CO2.

[0088] Analyses:

[0089] After 24 hours of incubation, the cells are lysed and treated in order to collect the protein fractions.

[0090] Protein concentrations are assessed by the BCA protein assay kit according to the standardized method (TEC-136).

[0091] The secretion of interleukin 6 is quantified by the ELIS A test using the “Quantikine® Human IL-6 Immunoassay” kit. The IL-6 concentration is expressed in “pg / pg of protein” based on the total amount of protein evaluated by the BCA test.

[0092] The analyses are repeated twice.

[0093] Results:

[0094] IL-6 production without UV-B irradiation for controls was assayed to validate the experiment. The results are shown in Table 2.

[0095] [TAB. 2] Compound Back calculation IL-6 (pg / ml) Mean pg / ml protein pg IL-6 / pg protein Mean Culture medium 9.87 9.49 + / -0.55 264.70 0.04 0.04 + / - 0.0 0 9.10 246.52 0.04 Ethanol 9.41 7.32 + / - 2.96 208.62 0.05 0.05 + / -0.00 5.23 114.49 0.05 Vit D3 8.02 8.41 + / -0.55 198.00 0.04 0.04 + / -0.00 8.79 245.00 0.04

[0096] The results of the IL-6 production assays with UV-B irradiation are presented in Tables 3 and 4, as well as in [Fig.2].

[0097] [TAB. 3] Sample Back calculation IL-6 (pg / ml) Mean pg / ml protein pg IL-6 / pg protein Mean Culture medium 21.85 20.63 + / -1.73 255.24 0.09 0.08 + / -0.01 19.40 263.72 0.07 Ethanol 24.75 24.45 + / -0.43 243.93 0.10 0.10 + / -0.00 24.14 245.34 0.10 Vit D3 14.34 13.88 + / -0.65 201.42 0.07 0.07 + / -0.01 13.42 211.35 0.06

[0098] [TAB. 4] Sample Back calculation IL- 6 ( pg / ml) Mean pg / ml protein pg IL-6 / pg protein Mean Hydrolysate 39.2 11.73 11.11 202.84 0.06 0.06 mg / ml 10.49 + / - 0.87 175.86 0.06 + / -0.00 Hydrolysate 7.84 12.19 11.88 265.13 0.05 0.05 mg / ml 11.57 + / -0.44 255.24 0.05 + / -0.00 Hydrolysate 1.57 12.96 12.19 269.37 0.05 0.05 mg / ml 11.42 + / -1.09 192.91 0.06 + / -0.01 Hydrolyzate 0.31 14.03 16.87 + / 217.02 0.06 0.07 mg / ml 19.71 -4.01 231.19 0.09 + / -0.01 Hydrolyzate 0.06 19.25 23.30 243.93 0.08 0.09 mg / ml 27.34 + / -5.72 249.59 0.11 + / -0.02

[0099] Interpretation:

[0100] It is noted that UV-B treatment does indeed induce an increase in IL-6 production (Table 2 versus Table 3).

[0101] Treatment with vitamin D3 does indeed decrease IL-6 production, compared with treatment with culture medium which serves as a reference: 13.8 + / - 0.65 compared to 20.63 + / - 1.73 pg / ml, and 0.07 + / - 0.01 compared to 0.08 + / - 0.01 pg / pg (See Table 3, and [Fig.2] on which the line indicates the reference, i.e. the level of IL-6 production under control conditions). Vitamin D3 is therefore a positive control.

[0102] The hydrolyzate according to the invention induces a decrease in the production of IL-6 at concentrations of 39.2 mg / ml (0.06 + / - 0.00 pg / pg compared to 0.08 + / - 0.01 pg / pg), 7.84 mg / ml (0.05 + / - 0.00 pg / pg compared to 0.08 + / - 0.01 pg / pg), 1.57 mg / ml (0.05 + / - 0.01 pg / pg compared to 0.08 + / - 0.01 pg / pg) and 0.31 mg / ml (0.07 + / - 0.01 pg / pg compared to 0.08 + / - 0.01 pg / pg) (See Table 4 and [Fig.2]).

[0103] Furthermore, the hydrolyzate according to the invention induces a reduction in the production of IL-6 greater than that of the positive control at concentrations of 39.2 mg / ml (0.06 + / -0.00 pg / pg compared to 0.07 + / - 0.01 pg / pg), 7.84 mg / ml (0.05 + / - 0.00 pg / pg compared to 0.07 + / - 0.01 pg / pg), 1.57 mg / ml (0.05 + / - 0.01 pg / pg compared to 0.07 + / - 0.01 pg / pg) (See Table 4 and [Fig.2]).

[0104] The hydrolyzate according to the invention does indeed exhibit anti-inflammatory activity.

[0105] Example 3: evaluation of the antioxidant properties of a hydrolysate according to the invention

[0106] The aim of this test is to evaluate the antioxidant properties of a hydrolysate according to the invention in accordance with the ORAC method ((acronym for Oxygen Radical Absorbance Capacity).

[0107] The hydrolyzate tested is obtained according to a process as described in Example 1.

[0108] The hydrolyzate is tested according to the following concentrations: 20 mg / ml; 10 mg / ml; 2 mg / ml; 1 mg / ml; 0.200 mg / ml; 0.100 mg / ml; 0.020 mg / ml; 0.010 mg / ml; 0.002 mg / ml and 0.001 mg / ml (75 mM phosphate buffer solvent pH 7.4).

[0109] Positive control: 3,4-dihydro-6-hydroxy-2,5,7,8-tetramethyl-2H-l-benzopyran-2-carboxylic acid (Trolox). Trolox is prepared at 2 mM (0.5 mg / ml) in 75 mM phosphate buffer pH 7.4 and diluted in the same buffer at concentrations: 1.0000 mM, 0.5000 mM, 0.2500 mM, 0.1250 mM and 0.0625 mM. 8 μl of each solution is added to each well to obtain the test concentrations of 50 pM, 25 pM, 12.500 pM, 6.250 pM and 3.125 pM (160 μl final volume).

[0110] Reagents: - KPO4 buffer, pH 7.4, at 75 mM. - 2,2'-Azobis(2-methylpropionamidine) dihydrochloride (AAPH) at 70 mM (18.98 mg / ml) in 75 mM phosphate buffer, pH 7.4. 16 pL are added to each well to obtain the test concentrations (final volume of 160 pL). AAPH is used as a source of peroxide radical. - Fluorescein (FL) prepared at 2.66 mM (1 mg / ml) in 75 mM phosphate buffer, pH 7.4, then diluted in the same buffer to a concentration of 0.8235 nM. 136 μl are added to each well to obtain a test concentration of 0.7 nM (final volume of 160 μL). [YES] Protocol

[0112] ORAC evaluation was performed according to the standard method (TEC-151). 136 μl of 0.8235 fluorescein and 8 μl of hydrolyzate sample, Trolox solution (positive control), or 75 mM phosphate buffer (negative control) were added to the appropriate wells. An additional control, without AAPH, containing 136 μl of 0.8235 fluorescein and 24 μl of phosphate buffer was performed. Each condition is duplicated.

[0113] The well plates were covered and incubated at 37°C for 15 min.

[0114] Reactions were initiated by adding 16 μl of 70 mM AAPH, except for wells without AAPH. The plates were covered and incubated at 37 °C for 60 min. The Fluorescence was measured every minute (excitation 450-490 nm, emission 510 and 530 nm) during incubation.

[0115] Determination of ORAC values

[0116] The consumption of fluorescein associated with its incubation in the presence of AAPH was estimated by measuring the fluorescence. A typical curve of fluorescence monitoring during the ORAC test was produced. The net area under the curve (Net-AUC) (AUC with hydrolyzate - AUC without hydrolyzate) is calculated and the result is standardized with respect to trolox.

[0117] Results

[0118] Table 5 illustrates the Net-AUC values ​​for Trolox based on the indicated doses.

[0119] [TAB. 5] Trolox Dose (g M) Net AUC 25,000 21,281 12,500 13,963 6,250 9,302 3,125 4,405 0.000 0.000

[0120] Four tested doses of the hydrolyzate according to the invention have Net-AUC values ​​in the range of Net-AUC values ​​of trolox and allow the calculation of Trolox equivalents ([Fig.3] Table 5): - the dose of 0.02 mg / ml is equivalent to 7.049 pM of Trolox - the dose of 0.01 mg / ml is equivalent to 4.150 pM of Trolox - the dose of 0.002 mg / ml is equivalent to 0.896 pM of Trolox - the dose of 0.001 mg / ml is equivalent to 0.571 pM of Trolox

[0121] The extrapolation of the data for 1 g and 100 g of hydrolyzate is also presented in Table 6. The protein hydrolyzate has on average a Trolox equivalent of 279,145,180 + / - 57,535,342 pM Trolox equivalent per 100 g of product. For comparison, cloves, one of the foods with the highest antioxidant activities, have a Trolox equivalent of 314,446 pM Trolox equivalent per 100 g of product. Thus, the protein hydrolyzate according to the invention has an antioxidant activity approximately 800 times higher than that of cloves.

[0122] [TAB.6] 4: Effects on the growth of pigs of a hydrolyzate according to the invention

[0123] At the start of the study, the pigs are approximately 8 weeks old and have entered the fattening period, which here lasts 15 weeks, until slaughter.

[0124] The pigs are divided into several batches:

[0125] Batch 1: pigs having received the hydrolysate according to the invention during the 2 weeks preceding slaughter, at a rate of 5g / day.

[0126] Batch 2: pigs having received the hydrolysate according to the invention during the 3 weeks preceding slaughter, at a rate of 5g / day.

[0127] Batch 3: pigs having received the hydrolysate according to the invention during the 6 weeks preceding slaughter, at a rate of 5g / day.

[0128] Batch 4: pigs having received the hydrolysate according to the invention for 6 weeks in the middle of the fattening period, at a rate of 5g / day.

[0129] Batch 5: pigs having received the hydrolysate according to the invention for 6 weeks from the start of the fattening period, at a rate of 5g / day.

[0130] For each batch, controls were followed, which did not receive the hydrolyzate according to the invention.

[0131] The animals are weighed at the beginning and at the end of the fattening period. The results are presented in Table 7. In this table are indicated for each batch the average daily weight gains ("ADG) for the pigs having received the hydrolysate ("Treated") and the pigs not having received the hydrolysate ("Controls"). The differences between the Treated and the Controls for each batch are indicated.

[0132] [TAB. 7] Number Initial weight (kg) GMQ (g / d) Difference (g) Batch 1 2 weeks end Controls 78 20.7 754 + 37 Treated 122 22.2 791 Batch 2 3 wk end Witnesses 225 21.9 782 + 57 Treated 185 22.8 839 Batch 3 6 wk end Witnesses 145 25.9 752 + 74 Treated 157 27.5 826 Batch 4 6 wk mid Witnesses 163 24.8 750 + 40 Treated 158 24.8 790 Batch 5 6 wk start Witnesses 176 22.9 768 + 5 Treated 125 25.0 773

[0133] A daily weight gain is observed in the animals having received the hydrolyzate for all the batches. However, it is noted that the daily weight gain increases when the hydrolyzate is distributed at the end of the fattening period: + 57g / day compared to the controls when it is distributed for the last 3 weeks (batch 2) and +74g / day compared to the controls when it is distributed for the last 6 weeks (batch 3). At the end of the fattening period, the pigs in batch 2 weigh on average 4 kg more than the controls and the pigs in batch 3 weigh on average 5 kg more than the controls.

Claims

Claims

1. Hydrolyzate of proteins from tissues or cartilages rich in fish collagen, obtained by enzymatic digestion using an endopeptidase enzyme of bacterial origin, said hydrolyzate being characterized in that it has a protein fraction having a molecular profile with the following distribution: - between 15 and 60% of peptides with a molecular weight of between 1000 and 5000 Da, - between 20 and 40% of peptides with a molecular weight of between 500 and 1000 Da, - between 10 and 40% of peptides with a molecular weight of between 150 and 500 Da, - between 1 and 10% of peptides with a molecular weight of less than 150 Da, and an aminogram in which, as a percentage of the total nitrogenous matter: - the glutamic acid content is between 10 and 14%, - the valine content is between 1.5 and 3.5% - the isoleucine content is between 1 and 3%, - the leucine content is between 2 and 6%,- the glycine content is between 9 and 13%, - the lysine content is between 0.5 and 3%, - the arginine content is between 6 and 10%.,

2. Hydrolyzate according to claim 1, characterized in that it also has: - a fat content of less than 0.5% as a percentage of dry extract, - a total nitrogenous matter content (N * 6.25) of greater than 90%, as a percentage of dry extract, - a mineral matter content of less than 10%, as a percentage of dry extract.

3. Hydrolysate according to claim 1 or 2, characterized in that the aminogram also presents, as a percentage of the total nitrogenous matter: - an aspartic acid content of between 7 and 10% - a proline content of between 7 and 11% - a threonine content is between 2 and 5% - a serine content is between 3 and 7% - an alanine content is between 8 and 12% - a cystine content is between 0.1 and 1% - a methionine content is between 1.5 and 3% - a tyrosine content is between 0.5 and 2.5% - a phenylalanine content is between 1 and 3.5% - a histidine content is between 3.5 and 7% - a hydroxyproline content is between 4 and 8% - a hydroxylysine content is between 0.5 and 2.5%.

4. Hydrolyzate according to one of the preceding claims, characterized in that said tissue is skin.

5. Hydrolyzate according to one of the preceding claims, characterized in that said fish is a Salmonidae, preferably salmon or trout, or a white fish.

6. Hydrolyzate according to one of the preceding claims, characterized in that said enzyme is the enzyme marketed under the brand name Corolase 7089.

7. Process for obtaining a fish protein hydrolysate as defined in one of claims 1 to 6, characterized in that it comprises the steps of: - preparation of a protein source from collagen-rich fish tissues and / or cartilages, - preparation of an enzymatic solution comprising between 0.1 and 0.5% of an endopeptidase enzyme, - enzymatic hydrolysis of said protein source at a temperature between 40 and 60°C, for 1 to 5 hours, after the addition of the enzymatic solution, - stopping the enzymatic hydrolysis by heating at a temperature of at least 90°C for 8 to 20 minutes, - separation of the protein hydrolysate from the other components, - recovery and concentration of the protein hydrolysate.

8. Method according to claim 7, characterized in that said enzyme is the enzyme marketed under the brand name Corolase 7089.

9. Method according to claim 7 to 8, characterized in that the protein source / water ratio is less than or equal to 1.

10. Method according to one of claims 7 to 9, characterized in that said tissue is skin.

11. Method according to one of claims 7 to 10, characterized in that said fish is a Salmonidae, preferably salmon or trout, or even a white fish.

12. Composition characterized in that it comprises a protein hydrolyzate as defined in one of claims 1 to 6, or obtained by a process as defined in one of claims 7 Q 1 1

13. d 11. Hydrolyzate of tissue or cartilage proteins rich in fish collagen as defined in one of claims 1 to 6, or obtained by a process as defined in one of claims 7 to 11, or composition containing it, for its use for stimulating the growth of farm animals, in particular pigs and poultry.

14. Hydrolyzate according to claim 13, characterized in that the growth stimulation comprises weight gain, in particular muscle gain.

15. Hydrolyzate of tissue or cartilage proteins rich in fish collagen as defined in one of claims 1 to 6, or obtained by a process as defined in one of claims 7 to 11, or composition containing it, for its use for the prevention and treatment of bursitis.

16. Hydrolyzate of proteins from tissues or cartilages rich in fish collagen as defined in one of claims 1 to 6, or obtained by a process as defined in one of claims 7 to 11, or composition containing it, for its use for the prevention and treatment of oxidation reactions and inflammatory reactions in farm animals, in particular pigs and poultry.

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