OA18348A - Composition containing chitin and digestible proteins - Google Patents

Abstract

The present invention relates to a composition comprising at least 67% by weight of crude protein, at least 5% by weight of chitin, the percentages by weight being given on the total weight of the composition, and 85% by weight of digestible proteins on the total weight of crude protein. It also relates to a process for preparing the composition as well as the uses thereof, in particular in human or animal food.

Description

COMPOSITION CONTAINING CHITINE AND DIGESTIBLE PROTEINS

The present invention relates to a composition comprising proteins and chitin. It also relates to a process for preparing this composition and its use in human or animal feed, and more particularly in fish feed.

Aquaculture is one of the most dynamic sectors of the food industry today. The high demand for fish has resulted in a significant increase in the price of feed for fish farming.

One of the most widely used products in fish feed is fish meal. Indeed, fishmeal is one of the main sources of protein in aquaculture feed. It is a flour very rich in animal proteins (rich in amino acids such as lysine and methionine) which are easy to digest. A growing demand accompanied by a limited supply has resulted in a significant increase in its price, posing a risk for the sustainable growth of aquaculture. Thus, there is a strong demand for alternative sources of high quality and, wherever possible, renewable protein for aquaculture feed.

Insect meal offers alternative natural protein sources and the possibility of being mass produced with a minimal ecological footprint. In particular, certain insects, such as Tenebrio molitor, have the advantage of being able to be adapted to intensive mass production.

However, the results of trials of substituting fishmeal with various insect meal have been mixed. If the substitution turns out to be possible, it does not generally exceed 50%, beyond this content adverse effects on the growth of fish are observed.

The work of the inventors made it possible to demonstrate that a specific composition could be advantageously used as a replacement for fish meal in aquaculture feed.

The present invention therefore relates to a composition comprising at least 67% by weight of crude protein, at least 5% by weight of chitin, the percentages by weight being given on the total weight of the composition, and 85% by weight of digestible proteins on the basis of. the total weight of crude protein.

It should be noted that in the context of this application, and unless otherwise specified, the ranges of values indicated are understood to be limits included.

The quantification of "crude protein" is well known to those skilled in the art. By way of example, we can cite the Dumas method or the Kjeldhal method. Preferably, the Dumas method, corresponding to standard NF EN ISO 16634-1 (2008) is used.

Throughout the application, where no date is specified for a regulation, standard or directive, it is the regulation, standard or directive in effect on the date of filing.

Preferably, the composition comprises 68% by weight of crude proteins, more preferably 70% by weight of crude proteins, the percentages by weight being given on the total weight of the composition.

The term “digestible proteins” refers to digestible proteins determined by the pepsi digestibility. The quantification of digestible proteins will preferably be carried out by the method described in Directive 72/199 / EC.

Preferably, the composition comprises 86%, more preferably 88% by weight of digestible proteins on the total weight of crude proteins.

According to the invention, by “chitin” is meant any type of chitin derivative, that is to say of polysaccharide derivative comprising N-acetyl-glucosamine units and D-glucosamine units, in particular chitin-polypeptide copolymers. (sometimes referred to as a "chitin-polypeptide composite"). These copolymers can also be combined with pigments, often of the melanin type.

Chitin is believed to be the second most synthesized polymer in the living world after cellulose. Indeed, chitin is synthesized by many species of the living world: it partly constitutes the exoskeleton of crustaceans and insects and the side wall that surrounds and protects fungi. More particularly, in insects, chitin thus constitutes 3 to 60% of their exoskeleton.

The determination of the chitin level is carried out by extraction thereof. One such method can be the ADAC 991.43 method described in Example 2, and is a preferred method for this determination.

Preferably, the composition comprises between 5 and 16% by weight of chitin, more preferably between 8 and 14% of chitin, the percentages by weight being given on the total weight of the composition.

The compositions of the prior art capable of containing both proteins and chitin are generally compositions obtained from insects and / or crustaceans. However, the high levels of crude protein and digestible protein in the composition according to the invention can only be obtained by a method for treating insects and / or crustaceans, comprising a hydrolysis step. However, a hydrolysis step has the effect of lowering the chitin level to a content of the order of 5% by weight, such as less than 5% by weight, on the total weight of the composition.

However, chitin is often considered as a kind of anti-nutritional factor because it is difficult to digest. This explains why for applications in the food industry, insect-based compositions are de-chitinated, that is to say a chitin removal step is performed. However, the work of the inventors has also made it possible to demonstrate that, contrary to popular belief, chitin had no impact on the growth of fish fed with a composition according to the invention, comprising a non-negligible level of chitin ( see Example 4 below). On the contrary, the composition according to the invention can advantageously replace, not only partially but also in its entirety, a fish meal in an aquaculture feed. Indeed, the composition according to the invention makes it possible to improve the growth of animals fed with this composition.

In addition, during the food manufacturing process, the introduction of the composition according to the invention also has certain advantages: reduction in losses of water-soluble vitamins during any heat treatment and reduction in the energy required during a possible extrusion step.

Preferably, the composition according to the invention has a residual humidity level of between 2 and 15%, preferably between 5 and 10%, more preferably between 6 and 8%. This humidity level can for example be determined according to the method resulting from the EC regulation 152/2009 of 27-01-2009 (103 ° C / 4 h).

Advantageously, the composition according to the invention has an ash content of less than or equal to 4% by weight on the total weight of the composition, and even more advantageously, less than or equal to 3.5%.

The ash constitutes the residue resulting from the combustion of the composition according to the invention.

The method of determining the ash content is well known to those skilled in the art. Preferably, the ashes were determined according to the method covered by regulation EC 152/2009 of 27-01-2009.

The fat content of the composition according to the invention is preferably between 5 and 20% by weight on the total weight of the composition, more preferably between 9 and 17%.

The methods for determining the fat content are well known to those skilled in the art. By way of example and preferably, the determination of this content will be carried out by following the method of Regulation EC 152/2009.

As indicated above, the composition according to the invention can be obtained from insects.

By "obtained" from insects, is meant more particularly a composition obtained only from insects and optionally water. The composition results from a mechanical, thermal treatment, excluding any chemical treatment (other than water) of the insects.

More particularly, the composition is an insect meal. By insect meal, we mean a powder having a particle size acceptable for human or animal consumption. The term “particle size acceptable for human or animal consumption” is intended to mean a particle size of between 100 μm and 1.5 mm, preferably between 300 μm and 1 mm, more preferably between 500 and 800 μm.

Preferred insects for the preparation of such a flour are, for example, Coleoptera, Diptera, Lepidoptera, Isoptera, Orthoptera, Hymenoptera, Blattoptera, Hemiptera, Heteroptera, Ephemeroptera and Mecoptera, preferably Coleoptera, Diptera, Orthoptera, Lepidoptera or their mixtures.

Preferably, the insects are chosen from the group consisting of Tenebrio molitor, Hermetia illucens, Galleria mellonella, Alphitobius diaperinus, Zophobas mono, Blattera fusca, Tribolium castaneum, Rhynchophorus ferrugineus, Musca domestica, Chrysomya megaceperchala, Schustoca migratoria Achregeta, Achregeta genus Achregeta, Samia ricini or their mixtures, and more preferably still, Tenebrio molitor.

Advantageously, the composition according to the invention comprises between 35 and 65% by weight of soluble proteins relative to the total weight of crude proteins, and at least 50% of the soluble proteins have a size less than or equal to 12400 g / mol.

By "soluble proteins" is meant, among crude proteins, those which

are soluble in an aqueous solution the pH of which is between 6 and 8, advantageously between 7.2 and 7.6.

Preferably, the aqueous solution is a buffer solution the pH of which is between 6 and 8, advantageously between 7.2 and 7.6. Preferably, the buffer solution is an NaCl phosphate buffer solution, the pH of which is equal to 7.4 +/- 0.2.

The digestibility of proteins in humans and animals is strongly influenced by the size of the proteins. In animal nutrition, it is common to reduce the size of proteins in order to facilitate digestion in animals. This reduction in the size of the proteins is generally carried out by hydrolysis processes (for example enzymatic), the implementation of which is particularly expensive.

The composition according to the invention, obtained by a process which does not involve hydrolysis, comprises a large amount of soluble proteins, the size of which is sufficiently small to facilitate digestion in animals. The composition according to the invention also has the advantage of being able to be prepared at a lower cost.

Advantageously, the composition according to the invention comprises between 30 and 60% by weight, preferably between 35 and 55% by weight of soluble proteins relative to the total weight of crude proteins.

Preferably, at least 60%, preferably at least 70% of the soluble proteins have a size less than or equal to 12400 g / mol.

More particularly, the soluble proteins have a size between 6500 and 12400 g / mol.

Advantageously, less than 10%, preferably less than 8%, more preferably less than 5%, and even more preferably 0% of the soluble proteins have a size greater than or equal to 66,000 g / mol.

Such a distribution of soluble proteins is demonstrated in Γ

Example 6.

The invention also discloses a process for preparing a composition according to the invention.

The process for preparing a composition according to the invention comprises a step of pressing the insects.

The objective of pressing is to de-oil the insects and therefore to obtain a press cake having an oil (or fat) content of less than or equal to 20% by weight on the dry weight of the press cake, preferably less or less. equal to 17%.

The pressing step is more fully described in step 2 of the process of

detailed preparation below.

In particular, it is possible to perform hot or cold pressing. Preferably, a single screw press is used.

More particularly, the preparation process according to the invention comprises the following steps:

i) slaughtering the insects, ii) pressing the insects to obtain a press cake, and iii) crushing the press cake.

The killing of insects can be carried out by scalding or bleaching, as is further described below in step 1 of the detailed process.

Likewise, the grinding is more fully described in step 4 of the detailed process.

Finally, the preparation process according to the invention may further include a step of drying the press cake.

The drying step is advantageously carried out after the pressing step and before the grinding step.

Drying is more fully described in step 3 of the detailed process.

Detailed process for preparing a composition according to the invention • Step 1: slaughtering the insects

This slaughter step 1 can advantageously be carried out by scalding or by bleaching. This step 1 kills the insects while lowering the microbial load (reducing the risk of spoilage and health) and by inactivating the internal enzymes of the insects that can trigger autolysis, and thus rapid browning of the insects.

For boiling, the insects, preferably larvae, are thus boiled in water for 2 to 20 min, preferably 5 to 15 min. Preferably, the water is at a temperature of between 95 to 105 ° C, preferably 100 ° C.

The quantity of water introduced during scalding is determined as follows: the ratio of the volume of water in mL to the weight in g of insect is preferably between 0.3 and 10, more preferably between 0 , 5 and 5, even more preferably between 0.7 and 3, even more preferably of the order of 1.

For bleaching, the insects, preferably larvae, are bleached by steam (nozzles or steam bed) at a temperature between 80 and 130 ° C, preferably between 90 and 120 ° C, more preferably between 95 and. 105 ° C,

preferably 98 ° C or else with water at a temperature between 95 and 105 ° C, preferably 100 ° C (by spray nozzles) or in mixed mode (water + steam) at a temperature between 80 and 130 ° C, preferably between 90 and 120 ° C, more preferably between 95 and 105 ° C, preferably 98 ° C. The residence time in the bleaching chamber is between 1 to 15 minutes, preferably between 3 and 7 minutes.

• Step (optional): grinding

The insects are removed from the scalding tank or the blanching chamber, they are then sieved (or drained), and placed in a crusher, such as a knife mill, to reduce the insects to particles.

To facilitate grinding, a quantity of water can be added. This amount of water is similar to that introduced during scalding step 1: the ratio of the volume of water in mL to the weight in g of insect is preferably between 0.3 and 10, more preferably between 0.5 and 5, even more preferably between 0.7 and 3, even more preferably of the order of 1. It is also possible to keep the scalding water and / or the water resulting from the bleaching for perform this step.

Preferably, after grinding, the size of the insect particles is less than 1 cm (largest particle size observable using a microscope), preferably less than 0.5 cm. Preferably, the size of the particles is between 300 μm and 3 mm, more preferably between 500 μm and 1 mm. It is not necessary to reduce the particle size excessively, for example to a size less than 250 µm.

• Step 2: pressing

The insects resulting from stage 1 of slaughter or the wet paste resulting from the optional grinding stage is then placed in a press according to an operating method which makes it possible to press and separate a juice comprising both an oily fraction and a protein fraction.

Preferably, the pressing step makes it possible to obtain a press cake comprising an oil content of less than or equal to 20% by weight on the dry weight of the press cake, preferably less than or equal to 17%, more preferably. still less than or equal to 15%.

Likewise, the pressing step makes it possible to obtain a press cake having a dry matter content of between 30% and 60%, preferably between 40% and 55%, and more preferably between 45% and 55%. %.

Any press system can be used to perform the pressing step, such as, for example, a single or twin screw press (Angel type twin screw press), a filter press (Choquenet type filter press ), a platen press, etc. These systems are well known to those skilled in the art who are able to determine the pressing conditions in order to obtain the oil and / or water contents mentioned above.

In particular, it is possible to perform hot or cold pressing. Advantageously, the pressing will be carried out hot, which makes it possible to increase the deoiling of the press cake. In particular, hot pressing makes it possible to obtain a press cake comprising an oil content of less than or equal to 17% by weight on the dry weight of the press cake, preferably less than or equal to 15%.

• Step 3: drying

The press cake is then dried by conventional technologies known to those skilled in the art. Drying can be direct or indirect (thin layer dryer, “paddle dryer”, “tubular dryer”, “disc-dryer”, etc.) at a temperature between 60 ° C and 200 ° C, for a period of 15 min to 24 hours. By way of example, the press cake can be placed and dried in ventilated / stirred air at a temperature between 80 and 100 ° C, preferably at 90 ° C for a period of between 3 and 7 hours, preferably 5 hours.

The objective of this drying step is to obtain a press cake having a moisture content of between 2 and 15%, preferably between 5 and 10%, more preferably still between 4 and 8%.

• Step 4: final grinding

The dried press cake is then placed in a mill, such as a hammer mill, to reduce the press cake to particles.

Advantageously, at the end of this final grinding, the size of the insect particles is less than 0.5 cm (largest particle size observable using a microscope), preferably of the order of 1 mm. More particularly, the particle size is between 300 μm and 1 mm, even more preferably between 500 and 800 μm.

The succession of these four steps makes it possible to obtain a composition according to the invention, comprising a high level of crude proteins and digestible proteins while maintaining a level of chitin of the order of at least 5% by weight on the weight. total 35 of the composition.

As indicated above, the pressing step can be carried out cold or hot.

By way of example of a process for obtaining a composition according to the invention, involving cold pressing:

Larvae, for example of T. molitor, are introduced into a beaker containing 200 ml of water previously brought to the boil, and slaughtered by boiling in a water bath at 100 ° C. After 5 minutes, the beaker is removed from the water bath, the larvae are drained, then mixed with a volume of water of 200 mL. The liquid thus obtained is passed through a twin-screw type press. The press cake thus obtained is dried for 24 hours in an oven at 70 ° C., then ground to 250 μm.

By way of example of a process for obtaining a composition according to the invention, involving hot pressing:

Larvae, for example of T. molitor, are introduced into a bleaching chamber and steam bleached for 5 min at 100 ° C. The larvae thus bleached are then introduced into a drying-type press suitable for products loaded with water. The press cake thus obtained is dried for 5 hours in an oven at 90 ° C., then ground in a hammer mill at 1 mm.

Preferably, the process for preparing a composition according to the invention comprises the following steps:

i) slaughtering the insects, ii) pressing the insects to obtain a press cake, iii) drying the press cake, and iv) crushing the press cake.

According to a first embodiment of the method according to the invention, the pressing step is preceded by a step of crushing the insects.

The invention therefore relates to a process for preparing a composition according to the invention comprising the following steps:

i) slaughtering the insects, ii) pressing the insects to obtain a press cake, iii) drying the press cake, and iv) crushing the press cake, in which the pressing step is preceded by a crushing step insects.

An advantage of the pre-pressing insect grinding step is more fully described in Example 5.

According to a second embodiment of the method according to the invention, the step of pressing the insects is carried out hot.

The invention therefore relates to a process for preparing a composition according to the invention comprising the following steps:

i) slaughtering the insects, ii) pressing the insects to obtain a press cake, iii) drying the press cake, and iv) crushing the press cake, in which the pressing step is carried out hot.

As indicated above, the hot pressing makes it possible to obtain a press cake comprising an oil content less than or equal to 17% by weight on the dry weight of the press cake, preferably less than or equal to 15%. .

According to a third embodiment of the method according to the invention, the step of crushing the press cake is carried out to a particle size of between 15,300 µm and 1 mm, preferably between 500 and 800 µm.

The invention therefore relates to a process for preparing a composition according to the invention comprising the following steps:

i) slaughtering the insects, ii) pressing the insects to obtain a press cake, iii) drying the press cake, and iv) crushing the press cake, in which the step of crushing the press cake is carried out at a particle size between 300 µm and 1 mm.

More particularly, in this third embodiment of the method according to the invention, the step of pressing the insects can be carried out hot. Alternatively, the pressing step could be preceded by a step of crushing the insects.

Finally, the invention relates to the use of a composition according to the invention in human or animal food.

Advantageously, the composition according to the invention can be used in the food of pets such as dogs, cats, birds, fish, reptiles, rodents.

More particularly, the composition according to the invention can be used in aquaculture (fish, crustaceans, molluscs, shellfish), the feed of poultry (chicken, turkey, game such as quail, pheasant, bustard), pigs, ruminants

(cattle, sheep, goats, horses), mink.

Finally, the composition according to the invention can be advantageously used as a replacement for a protein flour.

The term “protein flour” more particularly relates to fish meal, milk or whey powder, soybean concentrate flour (“CSP”), meat meal, such as for example of the poultry meal type ( "Poultry Meal").

The replacement can be partial or total.

Preferably, the composition according to the invention is used as a partial or total replacement for fishmeal, such as a 50 or 100% replacement.

Other characteristics and advantages of the invention will appear in the examples which follow, given by way of illustration, with reference to:

- Figure 1, which is a diagram illustrating the variations in water temperatures and dissolved oxygen levels in the tanks where the trout fed with different doses of composition according to the invention were reared,

- Figure 2, which comprises two diagrams illustrating the impact on the final body weight (Fig. 2A) and the consumption index (Fig. 2B) of trout fed with different doses of composition according to the invention.

- Figure 3, which illustrates the distribution of the lipids from the insect found in the juice and the press cake obtained by a process comprising a pressing step or a grinding and then pressing step, and

- Figure 4, which is a diagram representing the size exclusion chromatography analysis of the protein size of the composition according to the invention.

EXAMPLE 1: Process for preparing a composition according to the invention

The composition according to the invention is prepared from larvae of Tenebrio molitor.

On receipt of the larvae, the latter can be stored at 4 ° C for 0 to 15 days in their breeding tanks before slaughter without major degradation. The weight of the larvae (age) of the larvae used is variable and therefore their composition may vary, as shown in Table 1 below:

Biomass (Insects) mg 23 35 58 80 108 154 Dry matter % * 34 34 34.2 37.9 39.6 39.5 Ashes % * 1.59 1.52 1.6 1.75 1.67 1.43 Crude protein % * 22.6 22.2 22 23.2 23.1 23.2 Lipids % * 6.62 6.88 7.98 10.3 10.9 11.7 * The% are expressed in dry weight relative to the weight wet d e larvae.

Table 1: Biochemical composition of Tenebrio molitor larvae according to their weight.

• Step 1: Bleaching Insects

The live larvae (+ 4 ° C to + 25 ° C) are conveyed in a layer between 2 and 10 cm thick, on a perforated belt belt (1mm) to a bleaching chamber. The insects are thus bleached with steam (nozzles or steam bed) at 98 ° C or with water at 100 ° C (spray nozzles) or in mixed mode (water + steam). The residence time in the bleaching chamber is between 1 to 15 minutes, 10 ideally 5 minutes.

The temperature of the larvae at the end of the bleaching is between 75 ° C and 98 ° C.

• Step 2: Pressing

The larvae, once bleached, are conveyed to the feed hopper of a continuous single-screw press. The larvae during the passage in the press are maintained at a temperature above 70 ° C. in order to increase the deoiling yields. The principle of oil removal is to pressurize the material inside a cylindrical cage by means of an arrangement of screws and rings arranged on the central axis. The cage is lined on the inside with bars divided into sections and kept apart by spaces of different thicknesses following the work zone. The interstices thus formed allow the flow of an oil fraction and limit the passage of the so-called "dry" material, the protein fraction, which will be called "press cake", thereby participating in the pressurization.

The pressing yields obtained are between 48 and 55%.

Cake ⁼ (massecake / massera massecake)

The obtained press cake contains 35 to 40% dry matter, 67 to 75% protein and 13 to 17% fat, the percentages by weight being given on the dry weight of the press cake.

• Step 3: Drying

The press cake is then placed on a tray in a thin layer (approximately 2 cm) and is dried in ventilated air / stirred at 90 ° C for 5 hours in order to obtain a press cake having a dry matter content greater than 92 %.

This step helps to protect against any contamination that has occurred since slaughter.

The water activity (Aw) at the drying outlet is 0.35. The microbiological results show an absence of Salmonella spp (method: IRIS Salmonella BKR 23 / 07-10 / 11) and Enterobacteriaceae values less than 10 10 CFU / g (method: NF ISO 2128-2, December 2004, 30 ° C and 37 ° C).

• Step 4: Grinding

The dried press cake, mainly comprising proteins, is finally crushed using a continuous hammer mill (6 reversible spindles - 8 mm thick). The crusher is fed by a hopper with a flow rate adjustment flap 15 (180kg / h). The perforated grid used to control the particle size at the outlet is

0.8 mm. The motor rotation speed is 3000 rpm (electric motor, absorbed power 4 kW (5.5 HP)).

EXAMPLE 2: Characterization of the composition according to the invention

The composition prepared in Example 1 was characterized.

1. Analyzes

1.1 Determination of humidity

The humidity level is determined according to the method resulting from the EC regulation 152/2009 of 27-01-2009 (103 ° C / 4 h).

1.2 Determination of the amount of crude protein

Crude proteins are determined according to the so-called Dumas method, corresponding to standard NF EN ISO 16634-1 (2008).

1.3 Determination of the amount of chitin

The dietary fibers of insect flour are mainly composed of chitin, the latter was therefore determined according to the ADAC 991.43 method. The values thus obtained are therefore slightly overestimated.

1.4 Determination of the amount of fat

The fat was determined according to the method of EC regulation 152/2009.

1.5 Determination of the quantity of ash

The raw ashes were determined according to the method covered by regulation EC 152/2009 of 27-01-2009.

1.6 Determination of the amount of phosphorus

The phosphorus is determined by ICP (“induced coupled plasma”) with internal calibration.

1.7 Determination of energy

The energy value is obtained with the coefficients of the EU regulation 1169/201.

1.8 Determination of the amounts of amino acids and fatty acids

This determination was carried out by gas chromatography after hydrolysis and derivatization of amino acids and fatty acids respectively.

1.9 Determination of pepsi digestibility

Peptic digestibility is measured by the method described in the directive

72/199 / EC.

2. Results

The composition according to the invention is detailed in Table 2 below.

λ / '‘ | Macrommimen ^ | Humidity % * 5.32 Protein % * 67.09 Chitin % * 8.0 Fat % * 13.6 Ash % * 3.21 Total phosphorus % * 0.75 Energy MJ / kg 23.74

MST '. Arginine % * 2.56 Histidine % * 1.39 Isoleucine % * 2.11 Leucine % * 3.99 Lysine % * 3.32 Threonine % * 1.87 Valine % * 2.91

JW C12: 0 % * 0.03 C14: 0 % * 0.22 C15: 0 % * 0.01 C16: 0 % * 1.33 C16: 1 % * 0.05 C16: 1n-7 % * 0.16 C17: 0 % * 0.02 C17: 1 % * 0.01 C18: 0 % * 0.35 C18: 1n-9 % * 3.03 C18: 1n-7 % * 0.04 C18: 2n-6 % * 2.96 C18: 2tn-6 % * 0.02 C18: 3n-3 % * 0.14 C20: 0 % * 0.02 C20: 1n-9 % * 0.01

Methionine % * 1.43 C20: 2n-6% * 0.01 Cysteine % * 0.63 C22: 0% * 0.01 Phenylalanine % * 1.98 * The percentages by weight are expressed on the total weight of the composition. Tyrosine % * 2.68 Taurine % * 0.42 Aspartic acid + asparagine % * 4.51 Glutamic acid + glutamine % * 6.36 Alanine % * 3.83 Wisteria % * 2.54 Proline % * 3.18 Serine % * 2.94

Table 2: composition

Furthermore, a pepsi digestibility of 90 +/- 2% is obtained.

EXAMPLE 3: Alternative process for preparing a composition according to the invention

200 g of T. molitor larvae are introduced into a beaker, placed in a water bath at 100 ° C and containing 200 mL of water previously brought to the boil. After 5 minutes, the beaker is removed from the water bath, the larvae are drained, then mixed with a volume of water of 200 mL. The liquid thus obtained is passed through a twin-screw type press. The press cake thus obtained is dried for 24 hours in an oven at 70 ° C., then ground to 250 μm. A composition according to the invention is thus obtained.

EXAMPLE 4 Introduction of the composition according to the invention into fish feed

In the present example, the effect of the dietary inclusion of a composition according to the invention on growth, feed intake, feed conversion, body composition and apparent digestibility of nutrients in rainbow trout has been studied.

1. Materials and methods

1.1. Composition according to the invention

The composition used in this example is that obtained according to Example 1 and more fully described in Example 2.

1.2. Experimental regimes

A Fishmeal Diet (CTRL) has been formulated with convenient ingredients to meet the known nutritional needs of juvenile rainbow trout. This CTRL diet is 25% fishmeal, 8% 10 other protein sources of marine origin (squid meal and krill meal), while the remaining protein sources were soy protein concentrate, wheat gluten and corn gluten. On the basis of this formulation, four test regimens (Y5, Y7.5, Y15 and Y25) were formulated, in which the fishmeal was replaced by the composition according to the invention at respective rates of 20.30 , 60 and 15 100% (see Table 3 below).

Ingredients in% *: CTRL Y5 Y7.5 Y15 Y25 Fish meal LT70 ¹ 25.00 20.00 17.50 10.00 0.00 Krill flour ² 3.00 3.00 3.00 3.00 3.00 Squid flour ³ 5.00 5.00 5.00 5.00 5.00 Composition according to the invention 5.00 750 15.00 25.00 Soy protein concentrate ⁴ 14.00 14.00 14.00 14.00 14.00 Wheat gluten ⁵ 9.05 9.25 9.40 9.65 10.10 Corn gluten ⁶ 8.20 8.20 8.20 8.20 8.20 Soy flour 48 7.50 7.50 7.50 7.50 7.50 Whole peas 6.15 5.75 5.40 4.75 3.70 Fish oil 11.50 11.50 11.50 11.50 11.50 Colza oil 6.00 5.80 5.70 5.40 5.00 Pre-mix of vitamins and minerals ⁷ 1.50 1.50 1.50 1.50 1.50 Soy lecithin 1.00 1.00 1.00 1.00 1.00 Guar gum 0.20 0.20 0.20 0.20 0.20 Antioxidant 0.20 0.20 0.20 0.20 0.20 Sodium propionate 0.10 0.10 0.10 0.10 0.10 Mono Calcium Phosphate 1.30 1.70 2.00 2.60 3.50 DL-methionine 0.30 0.30 0.30 0.40 0.50 Yttrium oxide ⁸ 0.02 0.02 0.02 0.02 0.02 Dry matter (DM),% * 93.4 ± 0.0 93.1 ± 0.0 93, ± 0.1 95.0 ± 0.0 93.2 ± 0.0 Crude protein,% DM ** 48.5 ± 0.0 48.5 ± 0.1 48.5 ± 0.0 48.5 ± 0.0 48.5 ± 0.1 Fat 22.7 ± 0.2 22.7 ± 0.1 22.6 ± 0.2 22.7 ± 0.2 22.7 ± 0.2

Ingredients in% *: __ CTRL Y5__Y7,5__Y15__Y25

crude,% DM ** Ash,% DM ** 9.4 ± 0.0 8.8 ± 0.0 8.7 ± 0.1 8.1 ± 0.0 7.4 ± 0.0 Chitin,% DM ** 0.06 0.46 0.66 1.26 2.06 Gross energy, MJ / kg DM 23.2 ± 0.2 23.2 ± 0.0 23.2 ± 0.0 23.2 ± 0.1 23.2 ± 0.1

*% of dry matter relative to the total weight of the composition **% by dry weight relative to the total weight of dry matter ¹ Peruvian fish meal LT70: 71% crude protein (CP), 11% crude fat (MGB), EXALMAR, Peru; ² Krill meal: 61% PB, 19% MGB, Aker BioMarine Antarctic AS, Norway; ³ Super Prime without viscera: 82% PB, 3.5% MGB, Sopropêche, France; ⁴ Soycomil P: 62% PB, 0.7% MGB, ADM, the Netherlands; ⁵ VITEN: 84.7% PB. 1.3% MGB, ROQUETTE, France; ³ Corn gluten meal: 61% PB, 6% MGB, COPAM, Portugal; ⁷ PREMIX Lda, Portugal. Vitamins (IU or mg / kg diet): DL-alpha tocopherol acetate, 100 mg; Sodium Mnadlone Bisulfate, 25 mg; Retinyl acetate, 20,000 IU; DL-cholecalciferol, 2000 IU; thlamine, 30 mg; riboflavin, 30 mg; pyridoxine, 20 mg; cyanocobalamin, 0.1 mg; nlcotinic acid, 200mg; folic acid, 15 mg; ascorbic acid, 1000 mg; Inositol, 500 mg; blotin, 3 mg; calcium pantothenate, 100 mg; choline chloride, 1000 mg, betaine, 500mg. Minerals (g or mg / kg): cobalt carbonate, 0.65mg; copper sulfate, 9 mg; ferric sulfate, 6 mg; potassium lodide, 0.5 mg; manganese oxide, 9.6 mg; sodium selenlte, 0.01 mg; zinc sulfate, 7.5 mg; sodium chloride, 400 mg; calcium carbonate, 1.86 g; excipient wheat; ⁸ Yttrium oxide was incorporated in only a fraction of the feeds used for digestibility measurements.

Table 3: Formulation and composition of the experimental regimes.

Squid meal and krill levels were kept constant among all diets to ensure high palatability. Minor adjustments to the formulation of the diets tested were made to maintain the iso-nitrogenous (crude protein, 48.5% DM), isolipid (22.7% DM) and isoenergetic (crude energy, 23.2 MJ / kg DM) conditions. ). The levels of methionine and monocalcium phosphate supplementation in the tested diets were adjusted to match those found in the CTRL diet.

The schemes were manufactured by extrusion (pellet sizes: 1.2 and 2.0mm) by a pilot scale CLEXTRAL BC45 twin screw extruder with a screw diameter of 55.5mm and a temperature range of 119 to 123 ° vs. During extrusion, all of the extruded food batches were dried in a vibrating fluidized bed dryer (model DR100, TGC Extrusion, France). After cooling the granules, the oils were added by vacuum coating (model PG-10VCLAB, Dinnisen, The Netherlands). Throughout the test, the tested foods were stored at room temperature, but in a cool and ventilated place. Representative samples from each diet were taken for analysis (Tables

4-5).

lOTf KKCTR19 ⁷ --- "β" 0 »Y7i53SB TWT > -Th · .. * *> Arginine 4.62 ± 0.23 4.53 ± 0.02 4.49 ± 0.23 4.27 ± 0.09 3.89 ± 0.09 Histidine 1.47 ± 0.11 1.56 ± 0.02 1.54 ± 0.09 1.46 ± 0.07 1.50 ± 0.08 Isoleucine 2.31 ± 0.01 2.52 ± 0.01 2.53 ± 0.01 2.46 ± 0.02 2.49 ± 0.00 Leucine 4.51 ± 0.08 4.44 ± 0.01 4.68 ± 0.05 4.46 ± 0.02 4.56 ± 0.01 Lysine 3.09 ± 0.19 3.09 ± 0.01 3.02 ± 0.17 2.94 ± 0.01 2.97 ± 0.03 Threonine 2.32 ± 0.03 2.37 ± 0.00 2.31 ± 0.03 2.14 ± 0.05 2.15 ± 0.02 Valine 2.75 ± 0.00 2.87 ± 0.02 3.00 ± 0.03 3.08 ± 0.01 3.18 ± 0.01 Methionine 1.71 ± 0.15 1.71 ± 0.01 1.75 ± 0.06 1.74 ± 0.02 1.63 ± 0.02 Cysteine 0.35 ± 0.02 0.34 ± 0.00 0.31 ± 0.02 0.33 ± 0.00 0.34 ± 0.00 Phenylalanine 3.30 ± 0.00 3.06 ± 0.01 2.92 ± 0.15 2.85 ± 0.01 2.56 ± 0.00 Tyrosine 2.44 ± 0.11 2.48 ± 0.00 2.67 ± 0.14 2.92 ± 0.04 3.14 ± 0.12 Taurine 0.20 ± 0.01 0.20 ± 0.00 0.21 ± 0.01 0.06 ± 0.00 0.04 ± 0.00

The contents are indicated in percentages in inches on the total weight of granules before drying.

Table 4: Amino acid profile of the experimental diets.

C14: 0 0.40 ± 0.00 0.40 ± 0.00 0.38 ± 0.00 0.43 ± 0.00 0.38 ± 0.00 C16: 0 1.86 ± 0.01 1.89 ± 0.01 1.82 ± 0.02 2.11 ± 0.01 1.94 ± 0.02 C16: 1n-7 0.48 ± 0.00 0.48 ± 0.00 0.44 ± 0.00 0.50 ± 0.00 0.42 ± 0.01 C18: 0 0.49 ± 0.00 0.50 ± 0.01 0.47 ± 0.01 0.54 ± 0.00 0.50 ± 0.01 C18: 1n-9 1.62 ± 0.01 1.74 ± 0.01 1.69 ± 0.01 2.08 ± 0.01 2.06 ± 0.02 C18: 1n-7 0.26 ± 0.00 0.25 ± 0.00 0.23 ± 0.00 0.25 ± 0.00 0.21 ± 0.00 C18: 2n-6 0.79 ± 0.00 0.94 ± 0.01 1.05 ± 0.01 1.36 ± 0.01 1.53 ± 0.02 C18: 3n-3 0.13 ± 0.00 0.13 ± 0.00 0.13 ± 0.00 0.14 ± 0.00 0.12 ± 0.00 C18: 4n-3 0.10 ± 0.00 0.10 ± 0.00 0.09 ± 0.00 0.10 ± 0.00 0.08 ± 0.00 C20: 1n-9 0.20 ± 0.00 0.19 ± 0.00 0.17 ± 0.00 0.18 ± 0.00 0.14 ± 0.00 C20: 4n-6 0.14 ± 0.00 0.13 ± 0.00 0.12 ± 0.00 0.14 ± 0.00 0.12 ± 0.00 C20: 5n-3 0.72 ± 0.00 0.71 ± 0.01 0.65 ± 0.00 0.70 ± 0.00 0.57 ± 0.01 C22: 1n-11 0.14 ± 0.00 0.13 ± 0.00 0.11 ± 0.00 0.12 ± 0.00 0.08 ± 0.00 C22: 5n-3 0.14 ± 0.00 0.13 ± 0.00 0.12 ± 0.00 0.13 ± 0.00 0.10 ± 0.00 C22: 6n-3 1.45 ± 0.01 1.44 ± 0.01 1.33 ± 0.01 1.46 ± 0.01 1.21 ± 0.02

The contents are indicated as a percentage by weight of the total weight of granules before drying.

Table 5: Summary of the fatty acid profile of the experimental diets.

1.3. Growth performance test

Triplicate groups of 35 rainbow trout (Oncorhynchus mykiss), with an initial body weight (IBW) of 5.01 ± 0.1 g were fed one of the five experimental diets for 90 days. The fish were grown in 5 circular fiberglass tanks (volume: 250 L) supplied with continuously flowing fresh water, at temperatures between 14.1 ± 0.3 ° C and dissolved oxygen levels below. above 7.4 mg / L (see Figure 1). The fish were subjected by summer conditions to natural photoperiod changes (May-July). The fish were fed to apparent satiety, by hand, three times a day (9 a.m., 2 p.m. 10 and 6 p.m.) on weekdays and twice a day on weekends (10 a.m. and 4 p.m.), with the greatest care to avoid food waste. The food distributed was quantified throughout the study. Anesthetized fish were weighed individually at the beginning and at the end of the study and the group was weighed on day 28 and day 60. At the start, 15 fish from the same initial stock were sampled and stored at -20 ° C. for further analysis of full body composition. After 90 days of experimental feeding, 6 fish from each tank were sampled for the same purpose.

1.4. Apparent digestibility measurements

At the end of the growth test and following all associated sampling, fish (body weight: 45g) from each replicate tank were used to determine the apparent digestibility of dry matter, protein, fat, energy and phosphorus, by the indirect method with identical diets containing yttrium oxide (200 mg / kg) as inert tracer. The fish were stored in cylindrical-conical tanks (volume: 60 L; water flow rate: 3.7 L / min; dissolved oxygen levels greater than 6.4 mg / L), at a constant water temperature of 14 ° C. The fish were adapted for 10 days to the rearing conditions and to the experimental diets. Then, the fish were fed once a day (10:00 am), by hand in slight excess. After a thorough cleaning of the rearing tanks to remove all food residues, the faeces were collected daily for the next 8 days using the continuous filtration system of the outlet water (Choubert-INRA system ). After daily collection, the faeces were frozen at -20 ° C. The mixed faeces from each group of fish were lyophilized prior to analysis. Each diet has been tested in triplicate.

The apparent digestibility coefficients (ADCs) of nutrients and dietary energy in the experimental diets were calculated according to the formula:

CDA (%) = 100% ion concentrate ^{has "NEET%} concentration Υ2θ3 ^these ^% Energy or nutrients in faeces% Energy or nutrients in food

1.5. Analytical methods

Test ingredients, diets and freeze-dried feces were ground before analysis. The whole body samples were chopped, mixed, and a representative sample was lyophilized and homogenized with a laboratory mill before analysis. Analysis of the chemical composition of the ingredient, diets, feces and whole fish was done using the following procedures: dry matter after drying at 105 ° C for 24 h; ash by combustion at 550 ° C for 12 h; crude protein (N x 6.25) by a flash combustion technique followed by separation by gas chromatography and thermal conductivity detection (LECO FP428); fat by extraction with dichloromethane (Soxhlet); total phosphorus according to the ISO I DIS 6491 method using the vanado-moiybdic reagent; the raw energy in an adiabatic calorimeter bomb. Yttrium oxide in food and faeces was determined by the ICP-AES method.

For total amino acid analyzes, test ingredients and test diets were hydrolyzed (6 M HCL at 116 ° C for 22 h in nitrogen-rinsed glass vials), then derivatized with a fluorine reagent AccQ (6-aminoquinolyl-Nhydroxysuccinimidyle) according to the method of Tag AccQ (Waters, USA). Analyzes were performed by high performance liquid chromatography (HPLC) in a reverse phase amino acid analysis system, using norvaline as an internal standard. Tryptophan has not been determined because it is partially destroyed by acid hydrolysis. The resulting peaks were analyzed with EMPOWER software (Waters, USA). For the analysis of fatty acids, the lipids were extracted according to the method of Folch et al. (1957) and subsequently, the fatty acid composition of the fillets was determined by gas chromatography methyl ester analysis, according to the procedure of Lepage and Roy (1986).

1.6. Criterion for assessing growth and nutrient utilization

PCI (g): Initial body weight.

PCF (g): Final body weight.

Specific growth rate, TCS (% / day): (Ln PCF - Ln PCI) x 100 / days.

Consumption index, CI: gross food intake / weight gain.

Voluntary food intake, AVA (% CP / day): (gross food ration / (PCI + PCF) / 2 / days) x 100.

CEP protein efficiency coefficient: wet weight gain / crude protein intake.

Retention (% intake): 100 x (PCF x final carcass nutrient content - PCI x initial carcass nutrient content) / nutrient intake.

1.7. Statistical analysis

Data are presented by the average of three repetitions! the standard deviation.

The data were subjected to one-way analysis of variance. Before ANOVA, the values expressed in% were subjected to a transformation of the square root arc sine. Statistical significance was tested at a probability level of 0.05. All statistical tests were performed using IBM SPSSV21 software.

2. Results

2.1. Growth performance

Data on growth performance, feed conversion and protein efficiency of rainbow trout fed for 28, 60 and 90 days with the 25 experimental diets are reported in Tables 6-8 and Figure 2. No mortality occurred during the trial.

ISSOHi w ^- ' ·· PCI (g) 5.0 ± 0.1 4.9 ± 0.1 5.0 ± 0.1 5.1 ± 0.1 5.1 ± 0.1 PCF (g) 16.1 ± 0.1 ⁸ 16.2 ± 0.5 ⁸ 16.2 ± 0.5 ⁸ 17.9 ± 0.3 ^b 17.6 ± 0.4 ^b TCS,% / d 4.19 ± 0.12 ⁸ 4.26 ± 0.13 ⁸ 4.20 ± 0.07 ⁸ 4.50 ± 0.07 ^b 4.45 ± 0.06 ^b IC 0.87 ± 0.01 ^b 0.87 ± 0.02 ^b 0.87 ± 0.03 ^b 0.81 ± 0.00 ⁸ 0.81 ± 0.01 ⁸ Food intake,% CFM / d 3.27 ± 0.07 3.31 ± 0.09 3.28 ± 0.09 3.25 ± 0.03 3.22 ± 0.07 CEP 2.55 ± 0.02 ⁸ 2.56 ± 0.05 ⁸ 2.55 ± 0.08 ⁸ 2.66 ± 0.01 ^8b 2.72 ± 0.05 ^b

The values are the means ± the standard deviation (n = 3),

The values within a row with different exponents differ significantly (P <0.05).

Table 6: Growth performance on day 28.

After 28 days of experimental feeding (Table 5), the fish more than tripled their initial body weight. The food intake was high (3.22 - 3.31% CFM / day) and was not affected (P> 0.05) by the increasing doses of incorporation into the composition according to the invention. This observation suggests that the composition according to the invention had no negative effect on palatability and even that it could compensate for the complete elimination of fishmeal without compromising food intake. The growth rate ranged from 4.19 to 4.50% / day. Compared with CTRL treatment, while the Y5 and Y7.5 regimens did not affect PCF and TCS, the Y15 and Y25 regimens resulted in a significant (P <0.05) increase in PCF and TCS. The values of the consumption index vary between 0.81 and 0.87.

In comparison with CTRL, inclusion of the composition according to the invention at 5 and 7.5% (Y5 and Y7.5% regimes) did not affect the CI. However, the high inclusion levels of composition according to the invention (regimes of Y15 and Y25) led to a significant reduction in the CI (P <0.05). The protein efficiency coefficient (CEP) varied between 2.55 and 2.72. Fish fed the Y25 diet showed a significant increase in CEP, compared to those fed the CTRL, Y5 and Y7.5 diets.

W. Λ. PCI (g) 5.0 ± 0.1 4.9 ± 0.1 5.0 ± 0.1 5.1 ± 0.1 5.1 ± 0.1 PCF (g) 30.3 ± 0.1 ^a 31.6 ± 0.5 ’ 34.9 ± 1.5 " 37.2 ± 0.9 ^e 42.9 ± 0.4 ^d TCS,% / d 3.00 ± 0.04 ^a 3.10 ± 0.04 " 3.24 ± 0.04 ^e 3.31 ± 0.05 ^e 3.57 ± 0.04 ^d IC 1.10 ± 0.03 ^d 1.02 ± 0.03 ^e 0.92 ± 0.01 " 0.90 ± 0.02 " 0.85 ± 0.02 ^a CEP 2.01 ± 0.06 ^a 2.17 ± 0.06 " 2.40 ± 0.02 ^e 2.46 ± 0.06 " 2.56 ± 0.07 ^d

Values are means ± standard deviation (n = 3).

The values within a row with different exponents differ significantly (P <0.05).

Table 7: Growth performance at day 60.

After 60 days of experimental feeding (Table 6), the fish from the best performing treatment showed an 8-fold increase in initial body weight. The growth rate ranged from 3.00 to 3.57% / day. In comparison with the CTRL treatment, all the regimens with the composition according to the invention showed a significant increase (P <0.05) in TCS. CI values varied between

0.85 and 1.10 and in comparison with the CTRL, the inclusion of the composition according to the invention at all the doses tested resulted in a significant reduction in the CI (P <0.05). The protein efficiency coefficient (CEP) varied between 2.01 and 2.56. The lowest value of CEP was found in fish fed a CTRL diet, while improvement of CEP was closely associated with increasing doses of the composition according to the invention.

PCI (g) 5.0 ± 0.1 4.9 ± 0.1 5.0 ± 0.1 5.1 ± 0.1 5.1 ± 0.1 PCF (g) 42.9 ± 1.3 ^a 45.2 ± 1.0 ^b 49.0 ± 0.6 ^c 51.0 ± 1.4 ^e 55.9 ± 1.0 ^d TCS,% / d 2.39 ± 0.06 ^a 2.47 ± 0.02 ^b 2.54 ± 0.03 ^b 2.56 ± 0.05 ^b 2.67 ± 0.04 ^e IC 0.93 ± 0.02 ^b 0.83 ± 0.03 ^a 0.80 ± 0.02 ^a 0.79 ± 0.04 ^a 0.79 ± 0.02 ^a CEP 2.38 ± 0.06 ^a 2.68 ± 0.10 ^b 2.76 ± 0.06 ^b 2.80 ± 0.15 ^b 2.74 ± 0.08 ^b

Values are means ± standard deviation (n = 3).

The values within a row with different exponents differ significantly (P <0.05).

Table 8: Growth performance at day 90.

At the end of the trial, 90 days of experimental feeding (Table 7), the fish from the best performing treatment showed an 11-fold increase in initial body weight. In comparison with the CTRL fish, those fed the insect rich diets showed a significant increase in final body weight (P <0.05). This increase was dose related, with a moderate increase for the Y5 regimen, intermediate for Y7.5 and Y15, and the highest for Y25. The specific growth rate (TCS) varied between 2.39 and 2.67% / day, with a minimum value found in fish fed a CTRL diet, while those fed with feed containing composition according to The invention showed significantly higher TCS values (p <0.05). Regardless of the level of incorporation, the composition according to the invention led to a significant reduction in the CI (P <0.05). In comparison with CTRL treatment, all insect meal regimens led to a significant increase in CEP values (P <0.05).

2.2. Whole body composition

The data on the whole body composition of the trout at the end of the test are presented in Table 9. The feed treatments had no effect (P> 0.05) on the moisture content, protein, in fat, ash, phosphorus and whole fish energy.

CîHÜS. ··. ·. j- Humidity, % 70.1 ± 0.6 70.7 ± 0.4 71.1 ± 0.4 70.5 ± 0.5 70.7 ± 1.2 Protein,% 14.8 ± 0.6 14.8 ± 0.3 15.0 ± 0.5 15.2 ± 0.3 15.2 ± 0.7 Fat,% 12.2 ± 0.2 11.5 ± 0.4 11.0 ± 0.3 11.6 ± 0.1 11.8 ± 0.9 Ash,% 1.9 ± 0.0 2.2 ± 0.2 2.1 ± 0.3 2.1 ± 0.0 2.2 ± 0.1 Phosphorus,% 0.4 ± 0.0 0.4 ± 0.0 0.4 ± 0.0 0.4 ± 0.0 0.4 ± 0.0 Energy, kJ / g 8.2 ± 0.1 8.0 ± 0.0 8.0 ± 0.0 8.0 ± 0.2 8.2 ± 0.4

The percentages are percentages by weight of the total weight of the fish.

Values are means ± standard deviation (n = 3).

Initial fish: humidity 75.0%; protein 14.1%; fat 8.7%; ash 2.2%;

phosphorus 0.4%, energy 6.7 kJ / g.

Table 9: Composition of the whole body of trout fed with the various feed treatments.

2.3. Nutrient retention

The values of nutrients and energy retention (expressed as a percentage of the intake) are presented in Table 10. In comparison with the CTRL treatment, the fish fed with diets rich in the composition according to the invention showed a significant increase in protein and energy retention (P <0.05). Likewise, the Y7.5, Y15 and Y25 regimes showed significantly higher P retention than CTRL (P <0.05). Fat retention was not affected by diet (P> 0.05).

iSIRg— WWÎP Protein 35.5 ± 2.5 ^a 39.8 ± 0.7 ^b 41.6 ± 0.4 ^b 42.8 ± 2.2 ^b 41.9 ± 2.2 ^b Fat 64.4 ± 2.1 68.0 ± 4.9 66.8 ± 3.3 71.5 ± 3.4 70.9 ± 6.7 Phosphorus 30.5 ± 0.7 ^a 32.7 ± 1.8 ^ab 34.0 ± 0.7 ^b 33.9 ± 1.7 ^b 33.8 ± 1.1 ^b Energy 42.0 ± 0.8 ^a 45.4 ± 1.6 ^b 47.1 ± 1.4 ^b 47.8 ± 1.8 ^b 48.0 ± 2.9 ^b

Values are means ± standard deviation (n = 3).

The values within a row with different exponents differ significantly (P <0.05).

Table 10: Retention of nutrients and energy in trout fed by various diets.

2.4. Apparent digestibility

The composition of the faeces collected from the trout fed by the various feed treatments is presented in Table 11.

a.CompositionTdëM ¹ ,. T *, <i - · · t ‘i> Yttrium oxide, (mg / kg) 1384 ± 39 1395 ± 94 1415 ± 61 1369 ± 62 1411 ± 43 Protein,% DM * 19.63 ± 0.06 19.67 ± 0.24 19.76 ± 0.34 19.70 ± 0.38 19.20 ± 0.41 Fat,% DM * 4.37 ± 0.06 4.33 ± 0.19 4.28 ± 0.24 4.30 ± 0.06 4.20 ± 0.33 Phosphorus,% MS * 2.64 ± 0.06 2.77 ± 0.08 2.65 ± 0.10 2.54 ± 0.15 2.62 ± 0.09 Energy, kJ / g MS 23.24 ± 0.16 23.14 ± 0.40 23.47 ± 0.47 22.88 ± 0.16 23.09 ± 0.16

• Percentage by weight of the total weight of dry fecal matter.

Values are means ± standard deviation (n = 3).

Table 11: Composition of the faeces of trout fed with the various diets.

The apparent digestibility coefficients (CDA%) for the various nutrients and energy are presented in Table 12. The increase in the doses of incorporation of the composition according to the invention had no significant effect (P> 0 , 05) on the apparent digestibility of dry matter, protein, fat, phosphorus and energy.

ww - Dry matter 84.2 ± 0.4 84.2 ± 1.0 84.3 ± 0.7 84.0 ± 0.7 84.3 ± 0.5 Protein 93.6 ± 0.2 93.6 ± 0.4 93.6 ± 0.2 93.5 ± 0.4 93.8 ± 0.1 Fat 97.0 ± 0.1 97.0 ± 0.1 97.0 ± 0.2 97.0 ± 0.2 97.1 ± 0.3 Phosphorus,% intake 69.9 ± 1.4 68.3 ± 1.5 70.5 ± 2.4 71.4 ± 2.9 70.3 ± 1.8 Energy,% input 84.1 ± 0.4 84.3 ± 0.8 84.1 ± 1.0 84.2 ± 0.6 84.4 ± 0.6 The values are the means ± the standard deviation (n = 3).

Table 12: Apparent digestibility of nutrients and energy in trout.

3. Conclusion

At the end of the 90 days of experimental feeding, the overall growth performance can be considered very satisfactory and in a higher range for young rainbow trout, with TCS values for the total duration of the 5-month period. test varying between 2.4 and 2.7% / day. In the best performing treatments, the fish showed an 11-fold increase in their initial body weight. Feed conversion rates among treatments ranged between 0.79 and 0.93, suggesting good nutritional adequacy of the feeds and good feeding practices.

The experimental data generated in this example confirms that:

The incorporation of increasing doses of the composition according to the invention (5, 7.5, 15 and 25%) with a concomitant reduction in fishmeal has been progressively linked to a significant increase in the body weight of the fish.

All the diets containing the composition according to the invention showed a significant improvement in TCS, IC and CEP.

The increasing incorporation doses of the composition according to the invention had no effect on the whole body composition of the trout.

The increasing incorporation doses of the composition according to the invention had no effect on the apparent digestibility of dry matter, proteins, lipids, phosphorus and energy in the various experimental diets.

Proteins, phosphorus and energy retention were enhanced in trout fed with foods comprising the composition according to the invention.

In general, the composition according to the invention used in this example could effectively replace 100% of the fishmeal in the diet of juvenile rainbow trout with positive effects on CI and overall performance. of growth.

EXAMPLE 5 Processes with or without grinding prior to pressing

Press-only process

200 g of T. molitor larvae are introduced into a beaker, placed in a water bath at 100 C and containing 200 mL of water previously brought to the boil. After 5 minutes, the beaker is removed from the water bath, the larvae are drained, then passed through a twin-screw type press. A press cake is thus obtained.

Process with grinding followed by pressing

200 g of T. molitor larvae are introduced into a beaker, placed in a water bath at 100 ° C and containing 200 mL of water previously brought to the boil. After 5 minutes, the beaker is removed from the water bath, the larvae are drained, then mixed with a volume of water of 200 mL. The liquid thus obtained is passed through a twin-screw type press. A press cake is thus obtained.

Lipid level measurement

Place 2 g of sample in a beaker, add 0.2 g of Na2SO4 and 15 mL of CHClb / MeOH (2/1 v / v). The whole is placed under magnetic stirring for 20 minutes, then the solution is filtered, the residue is placed again in the beaker with 10 mL of CHCh / MeOH (2/1 v / v). The whole is placed under magnetic stirring for 15 minutes, then the solution is filtered, the solvent phases are combined and evaporated to constant weight. The lipid content is determined as a percentage of mass after extraction-evaporation relative to the initial mass of the sample (2 g).

Conclusion:

The importance of grinding upstream of pressing was studied (Figure 3). It thus clearly appears that the distribution of lipids between the cake and the press juice is much more efficient, 12.9 versus 87.1 versus 42.7 versus 57.3, when prior grinding has been carried out.

EXAMPLE 6 Analysis of the size of the soluble proteins of the composition according to the invention.

A 100 mg sample of the composition prepared in Example 1 was placed in 10 mL of NaCl phosphate buffer (pH 7.4, 0.137 mM). The sample was shaken for 1 minute (vortex), then centrifuged at 900 xg for 1 min. Following centrifugation, the sample was filtered through a 0.45 µm membrane. The size analysis of soluble proteins was performed using a size exclusion chromatography system, with a Nucleogel GFC-300 column. NaCl phosphate buffer (pH 7.4, 0.137 mM) was used as the eluent. The flow rate was 1.0 mL / min. The detection was carried out by a UV detector at 280 nm.

The results of the analysis are shown in Figure 4 and summarized in Table 13 below.

Protein size (kg / mol) Relative abundance (%) 6.5 to 12.4 74.4 12.4 to 29 20.5 29 to 66 5.1 Table 13: Distribution of tai the soluble proteins contained in the

composition prepared in Example 1

The results show that approximately 74.4% of the soluble proteins present in the composition according to the invention have a molar mass of less than 12400 g / mol (or Da, Daltons)

Claims (13)

1. Composition comprising at least 67% by weight of crude proteins, at least 5% by weight of chitin, the percentages by weight being given on the total weight of the composition, and 85% by weight of digestible proteins on the total weight of proteins raw. 2. Composition according to claim 1, comprising ash in a content of less than or equal to 4% by weight on the total weight of the composition. 3. Composition according to claim 1 or 2, comprising fat in a content of between 5 and 20% by weight on the total weight of the composition. 4. Composition according to any one of claims 1 to 3, obtained at 15 from insects. 5. Composition according to any one of claims 1 to 4, in which the residual moisture content is between 2 and 15%. 20 6. Composition according to any one of claims 1 to 5, comprising between 30 and 60% by weight of soluble proteins relative to the total weight of crude proteins, in which at least 50% of the soluble proteins have a size less than or equal to 12400g / mol. 25 7. A process for preparing a composition according to any one of claims 4 to 6, comprising the following steps: i) slaughtering the insects, ii) pressing the insects to obtain a press cake, and iii) crushing the press cake. 8. The method of claim 7, further comprising a step of drying the press cake. 9. The method of claim 8, comprising the following steps, i) slaughtering the insects, ii) pressing the insects to obtain a press cake, iii) drying the press cake, and iv) crushing the press cake, in which the pressing step is preceded by a crushing step 5 insects. 10. The method of claim 8, comprising the following steps, i) slaughtering the insects, ii) pressing the insects to obtain a press cake, iii) drying the press cake, and Iv) grinding the press cake, in which the pressing step is carried out hot. 11. The method of claim 8, comprising the following steps, i) slaughtering the insects, ii) pressing the insects to obtain a press cake, Iii) drying the press cake, and iv) grinding the press cake, wherein the step of crushing the press cake is carried out to a particle size between 300 µm and 1 mm. 12. Use of the composition according to any one of claims 1 to 6, 20 in food or feed. 13. Use according to claim 12, in which the composition is used as a replacement for a protein flour.