BEETLE POWDER

Abstract

The present invention relates to a beetle powder containing at least 67% by weight protein and at least 5% by weight chitin, the weight percentages relating to the total weight of the beetle powder. The invention is also directed to a method for preparing the beetle powder and to the uses thereof, in particular in human or animal nutrition.

Claims

1. Beetle powder comprising at least 67% by weight proteins and at least 5% by weight chitin, the percentages by weight being given relative to the total weight of beetle powder.

2. Beetle powder according to claim 1, comprising ash in a content less than or equal to 4% by weight relative to the total weight of beetle powder.

3. Beetle powder according to claim 1 or 2, comprising fat in a content comprised between 5 and 20% by weight relative to the total weight of beetle powder.

4. Beetle powder according to any one of claims 1 to 3, the proteins of which exhibit a digestibility greater than or equal to 85%.

5. Beetle powder according to any one of claims 1 to 4, the residual moisture content of which is comprised between 2 and 15%.

6. Beetle powder according to any one of claims 1 to 5, comprising between 40 and 60% by weight soluble proteins relative to the total weight of proteins, in which at least 50% of the soluble proteins have a size less than or equal to 12,400 g/mol.

7. Method for the preparation of a beetle powder according to any one of claims 1 to 6, comprising the following steps: i) killing the beetles, ii) pressing the beetles in order to obtain a press cake, and iii) grinding the press cake.

8. Method according to claim 7, also comprising a step of drying the press cake.

9. Method according to claim 8, comprising the following steps: i) killing the beetles, ii) pressing the beetles in order to obtain a press cake, iii) drying the press cake, and iv) grinding the press cake, in which the pressing step is preceded by a step of grinding the beetles.

10. Method according to claim 8, comprising the following steps: i) killing the beetles, ii) pressing the beetles in order 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. Method according to claim 8, comprising the following steps: i) killing the beetles, ii) pressing the beetles in order to obtain a press cake, iii) drying the press cake, and iv) grinding the press cake, in which the step of grinding the press cake is carried out to a particle size comprised between 300 μm and 1 mm.

12. Use of the beetle powder according to any one of claims 1 to 6, in human or animal nutrition.

13. Use according to claim 12, in which the beetle powder is used to replace protein flour.

Description

[0118] Other features and advantages of the invention will become apparent from the following examples, given by way of illustration, with reference to:

[0119] FIG. 1, which is a diagram illustrating the variations in temperatures of the water and in the levels of oxygen dissolved in the tanks where trout fed with different doses of beetle powder according to the invention were farmed,

[0120] FIG. 2, which comprises two diagrams illustrating the impact on final body weight (FIG. 2A) and the feed conversion ratio (FIG. 2B) of trout fed with different doses of beetle powder according to the invention,

[0121] FIG. 3, which illustrates the distribution of the lipids originating from the insect found in the juice and the press cake obtained by a method comprising a pressing step or a grinding step then pressing,

[0122] FIG. 4, which is a diagram representing the steric exclusion chromatography analysis of the size of the proteins of the beetle powder according to the invention.

EXAMPLE 1: METHOD FOR THE PREPARATION OF A BEETLE POWDER ACCORDING TO THE INVENTION

[0123] The beetles used for preparing the beetle powder are Tenebrio molitor larvae. Upon receipt of the larvae, they can be stored at 4° C. for 0 to 15 days in their rearing tanks without major degradation before being killed. The weight (age) of the larvae used is variable and as a result their composition can vary, as illustrated in Table 1 below:

TABLE-US-00001 TABLE 1 Biochemical composition of Tenebrio molitor larvae according to their weight. Biomass (insects) mg 23 35 58 80 108 154 Dry matter %* 34 34 34.2 37.9 39.6 39.5 Ash %* 1.59 1.52 1.6 1.75 1.67 1.43 Crude proteins %* 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 as dry weight relative to the wet weight of larvae.

[0124] Step 1: Blanching of the Insects

[0125] Living larvae (+4° C. to +25° C.) are conveyed in layers with a thickness comprised between 2 and 10 cm, on a perforated conveyor belt (1 mm) to a blanching chamber. The insects are thus blanched in steam (steam nozzles or bed) at 98° C. or in water at 100° C. (spray nozzles) or in mixed mode (water+steam). The residence time in the blanching chamber is comprised between 1 to 15 minutes, ideally 5 min.

[0126] The temperature of the larvae after blanching is comprised between 75° C. and 98° C.

[0127] Step 2: Pressing

[0128] The larvae, once blanched, are conveyed to the feed hopper of a continuous single-screw press. While passing into the press, the larvae are maintained at a temperature above 70° C. in order to increase the de-oiling yields. The principle of de-oiling is to pressurize the material inside a cylindrical cage by means of an arrangement of screws and rings arranged on the central shaft. The cage is lined inside with bars distributed in sections and kept apart by spaces of different thicknesses depending on the work area. The interstices thus arranged allow the flow of an oil/fat fraction while limiting the passage of the so-called “dry” matter, the protein fraction, which is called “press cake”, thus being involved in the pressurization.

[0129] The pressing yields obtained are comprised between 48 and 55%.

Y.sub.cake=(mass.sub.cake/mass.sub.juice+mass.sub.cake)

[0130] The press cake obtained contains 35 to 40% dry matter, 67 to 75% proteins and 13 to 17% fats, the percentages by weight being given relative to the dry weight of press cake.

[0131] Step 3: Drying

[0132] The press cake is then arranged on a tray in a thin layer (approximately 2 cm) and is dried in ventilated/stirred air at 90° C. for 5 hours in order to obtain a press cake having a dry matter content greater than 92%.

[0133] This step makes it possible to guard against any contamination having occurred since the killing.

[0134] The Aw (water activity) after drying is 0.35. The microbiological results show an absence of Salmonella spp (method: IRIS Salmonella BKR 23/07-10/11) and Enterobacteria values less than 10 CFU/g (method: NF ISO 2128-2, December 2004, 30° C. and 37° C.).

[0135] Step 4: Grinding

[0136] The dried press cake, comprising mainly proteins, is finally ground using a continuous hammer mill (6 reversible moving parts—thickness 8 mm). The grinder is fed by a hopper with a flow rate control flap (180 kg/h). The perforated grill used to control the output granulometry is 0.8 mm. The speed of rotation of the motor is 3000 rpm (electric motorization, absorbed power 4 kW (5.5 CV)).

EXAMPLE 2: CHARACTERIZATION OF THE BEETLE POWDER ACCORDING TO THE INVENTION

[0137] The beetle powder prepared in Example 1 was characterized.

[0138] 1. Analyses

[0139] 1.1 Determination of the Moisture Content

[0140] The moisture content is determined according to the method originating from EC Regulation 152/2009 of 27 Jan. 2009 (103° C./4 h).

[0141] 1.2 Determination of the Quantity of Crude Proteins

[0142] The crude proteins are determined according to the method called Dumas, and corresponding to the standard NF EN ISO 16634-1 (2008).

[0143] 1.3 Determination of the Quantity of Chitin

[0144] The dietary fibres in the insect meal are essentially composed of chitin, the latter was therefore assayed according to the method AOAC 991.43. The values thus obtained are slightly overestimated as a result.

[0145] 1.4 Determination of the Quantity of Fat

[0146] The fat was determined according to the method of EC Regulation 152/2009.

[0147] 1.5 Determination of the Quantity of Ash

[0148] The crude ash was determined according to the method under EC Regulation 152/2009 of 27 Jan. 2009.

[0149] 1.6 Determination of the Quantity of Phosphorus

[0150] The phosphorus is assayed by ICP (“induced coupled plasma”) with internal calibration.

[0151] 1.7 Determination of Energy

[0152] The energy value is obtained with the coefficients of EU Regulation 1169/2011.

[0153] 1.8 Determination of the Quantities of Amino Acids and Fatty Acids

[0154] This determination was carried out by gas chromatography after hydrolysis and derivatization of the amino acids and fatty acids respectively.

[0155] 1.9 Determination of Pepsin Digestibility

[0156] The pepsin digestibility is measured by the method described in Directive 72/199/EC.

[0157] 2. Results

[0158] The composition of this beetle powder is presented in Table 2 below.

TABLE-US-00002 TABLE 2 Composition of the beetle powder. Unit Powder Macronutrient Moisture %* 5.32 Protein %* 67.09 Chitin %* 8.0 Fat %* 13.6 Ash %* 3.21 Total phosphorus %* 0.75 Energy MJ/kg 23.74 Amino acids Arginine %* 2.56 Histidine %* 1.39 Isoleucine %* 2.11 Leucine %* 3.99 Lysine %* 3.32 Threonine %* 1.87 Valine %* 2.91 Methionine %* 1.43 Cysteine %* 0.63 Phenylalanine %* 1.98 Tyrosine %* 2.68 Taurine %* 0.42 Aspartic acid + %* 4.51 asparagine Glutamic acid + %* 6.36 glutamine Alanine %* 3.83 Glycine %* 2.54 Proline %* 3.18 Serine %* 2.94 Fatty acids 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 C20:2n-6 %* 0.01 C22:0 %* 0.01 *The percentages by weight are expressed relative to the total weight of powder.

[0159] Moreover, a pepsin digestibility of 90+/−2% is obtained.

EXAMPLE 3: ALTERNATIVE METHOD FOR THE PREPARATION OF A BEETLE POWDER ACCORDING TO THE INVENTION

[0160] 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 brought to the boil beforehand. 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 into a press of the twin-screw type. The press cake thus obtained is dried for 24 hours in an oven at 70° C., then ground to 250 μm. A beetle powder is thus obtained.

EXAMPLE 4: INTRODUCTION OF THE BEETLE POWDER ACCORDING TO THE INVENTION INTO FISH FEED

[0161] In the present example, the effect of including a beetle powder in feed on growth, feed intake, feed conversion, body composition and the apparent digestibility of the nutrients in the rainbow trout was studied.

1. Material and Methods

[0162] 1.1. Beetle Powder

[0163] The beetle powder utilized in this example is that obtained according to Example 1 and described more fully in Example 2.

[0164] 1.2. Experimental Diets

[0165] A fishmeal-based diet (CTRL) was formulated with convenient ingredients in order to meet the known nutritional needs of juvenile rainbow trout. This CTRL diet is composed 25% of fishmeal, 8% of other protein sources of marine origin (squid meal and krill meal), while the remaining protein sources were a concentrate of soy protein, wheat gluten and maize gluten. On the basis of this formulation, four test diets (Y5, Y7.5, Y15 and Y25) were formulated, in which the fishmeal was replaced with beetle powder in respective contents of 20, 30, 60 and 100% (see Table 3 below).

TABLE-US-00003 Ingredients in %*: CTRL Y5 Y7.5 Y15 Y25 Fishmeal LT70.sup.1 25.00 20.00 17.50 10.00 0.00 Krill meal.sup.2 3.00 3.00 3.00 3.00 3.00 Squid meal.sup.3 5.00 5.00 5.00 5.00 5.00 Beetle powder 5.00 750 15.00 25.00 Concentrate of soy 14.00 14.00 14.00 14.00 14.00 proteins.sup.4 Wheat gluten.sup.5 9.05 9.25 9.40 9.65 10.10 Maize gluten.sup.6 8.20 8.20 8.20 8.20 8.20 Soy meal 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 Rapeseed oil 6.00 5.80 5.70 5.40 5.00 Pre-mixture of vitamins 1.50 1.50 1.50 1.50 1.50 and minerals.sup.7 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 Monocalcium 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.sup.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 Crude fats, % DM** 22.7 ± 0.2 22.7 ± 0.1 22.6 ± 0.2 22.7 ± 0.2 22.7 ± 0.2 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 of 23.2 ± 0.2 23.2 ± 0.0 23.2 ± 0.0 23.2 ± 0.1 23.2 ± 0.1 DM *% of dry matter relative to the total weight of the composition **% by dry weight relative to the total weight of the dry matter

[0166] Table 3: Formulation and Composition of the Experimental Diets.

[0167] The levels of squid and krill meal were kept constant among all the diets in order to guarantee a high palatability. Minor adjustments were made to the formulation of the diets tested in order to maintain the isonitrogenous conditions (crude protein, 48.5% DM), isolipidic conditions (22.7% DM) and isoenergetic conditions (crude energy, 23.2 MJ/kg DM). The levels of supplementation with methionine and monocalcium phosphate in the diets tested were adjusted in order to correspond to those found in the CTRL feed.

[0168] The diets were produced by extrusion (granule sizes: 1.2 and 2.0 mm) using a CLEXTRAL BC45 twin-screw extruder on a pilot scale with a screw diameter of 55.5 mm and a temperature range of 119 to 123° C. During the extrusion, all the batches of extruded feeds were dried in a vibrating fluidized bed dryer (model DR100, TGC Extrusion, France). After cooling the granules, the oils were added by coating under vacuum (model PG-10VCLAB, Dinnisen, Netherlands). Throughout the duration of the test, the experimental feeds were stored at ambient temperature, but in a cool, well-ventilated place. Samples representative of each diet were taken for analysis (Tables 4-5).

TABLE-US-00004 Amino acids CTRL Y5 Y7.5 Y15 Y25 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

[0169] The contents are indicated in % by weight relative to the total weight of granules before drying.

TABLE-US-00005 TABLE 4 Amino acid profile of the experimental diets. Fatty acids CTRL Y5 Y7.5 Y15 Y25 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

[0170] The contents are indicated in % by weight relative to the total weight of granules before drying.

[0171] Table 5: Synthesis of the Fatty Acid Profile of the Experimental Diets.

[0172] 1.3. Growth Performance Test

[0173] Triplicate groups of 35 rainbow trout (Oncorhynchus mykiss), with an initial body weight (IBW) of 5.01±0.1 g were fed with one of the five experimental diets for 90 days. The fish grew in circular glass-fibre tanks (volume: 250 L) supplied with a continuous flow of fresh water at temperatures comprised between 14.1±0.3° C. and levels of dissolved oxygen above 7.4 mg/L (see FIG. 1). The fish were subjected to summer conditions with natural photoperiod changes (May-July). The fish were hand-fed to apparent satiation, three times a day (9 h00, 14 h00 and 18 h00) during the week and twice a day at week-ends (10 h00 and 16 h00), taking the greatest care to avoid wasting the feed. The feed distributed was quantified throughout the length of the study. Anaesthetized fish were weighed individually at the start and at the end of the study and the group was weighed on day 28 and on day 60. At the start, 15 fish of the same initial stock were sampled and stored at −20° C. for subsequent analysis of the whole body composition. After 90 days of experimental feeding, 6 fish from each tank were sampled for the same purpose.

[0174] 1.4. Apparent Digestibility Measurement

[0175] At the end of the growth test and following all the associated samplings, 12 fish (body weight: 45 g) from each replica tank were used to determine the apparent digestibility of the dry matter, proteins, lipids, energy and phosphorus, by the indirect method with identical diets containing yttrium oxide (200 mg/kg) as inert tracer. The fish were stored in cylindro-conical tanks (volume: 60 L; water flow rate: 3.7 L/min; levels of dissolved oxygen greater than 6.4 mg/L), at a constant water temperature of 14° C. The fish were adapted to the farming conditions and to the experimental diets over 10 days. Then, the fish were hand-fed once a day (10 h00), to slight excess. After deep cleaning of the rearing tanks to remove all the feed residues, the faecal matter was collected daily for the following 8 days using the continuous outlet water filtration system (Choubert-INRA system). After daily collection, the faecal matter was frozen at −20° C. The mixed faecal matter originating from each group of fish was lyophilized before analysis. Each diet was tested in triplicate.

[0176] The apparent digestibility coefficients (ADC) of the nutrients and of the feed energy in the experimental diets were calculated according to the formula:

[00001]$\mathrm{ADC}\left(\%\right)=100-\hspace{1em}\left[\frac{\%.Math..Math.\mathrm{concentration}.Math..Math.Y_2.Math.O_3.Math..Math.\mathrm{feed}}{\%.Math..Math.\mathrm{concentration}.Math..Math.Y_2.Math.O_3.Math..Math.\mathrm{faeces}}\times \frac{\%.Math..Math.\mathrm{Energy}.Math..Math.\mathrm{or}.Math..Math.\mathrm{nutrients}.Math..Math.\mathrm{in}.Math..Math.\mathrm{faeces}}{\%.Math..Math.\mathrm{Energy}.Math..Math.\mathrm{or}.Math..Math.\mathrm{nutrients}.Math..Math.\mathrm{in}.Math..Math.\mathrm{feed}}\right]$

[0177] 1.5. Analytical Methods

[0178] The test ingredients, the diets and the lyophilized faecal matter 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. The analysis of the chemical composition of the ingredient, diets, faecal matter and whole fish was carried out 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×6.25) by a flash combustion technique followed by separation by gas chromatography and thermal conductivity detection (LECO FP428); the fat by extraction with dichloromethane (Soxhlet); the total phosphorus according to the ISO/DIS 6491 method using vanadomolybdic reagent; the crude energy in a adiabatic bomb calorimeter. The yttrium oxide in the feeds and the faeces was determined by the ICP-AES method.

[0179] For analyses of total amino acids, the test ingredients and the test diets were hydrolysed (6 M of HCL at 116° C. for 22 h in glass flasks rinsed with nitrogen), then derivatized with an AccQ (6-aminoquinolyl-N-hydroxysuccinimidyl) fluorine reagent according to the AccQ-Tag method (Waters, USA). The analyses were carried out by high performance liquid chromatography (HPLC) in a reverse-phase amino acid analysis system, using norvaline as internal standard. The tryptophan was not determined as it is partially destroyed by acid hydrolysis. The resulting peaks were analysed with EMPOWER software (Waters, USA). For the analysis of the 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 analysis of the methyl esters by gas chromatography, according to the Lepage and Roy procedure (1986).

[0180] 1.6. Criterion for Evaluating Growth and Use of the Nutrients

[0181] IBW (g): Initial body weight.

[0182] FBW (g): Final body weight.

[0183] Specific growth rate, SGR (%/day): (Ln FBW−Ln IBW)×100/days.

[0184] Feed conversion ratio, FCR: gross feed ration/weight gain.

[0185] Voluntary feed intake, VFI (% BW/day): (gross feed ration/(IBW+FBW)/2/days)×100.

[0186] Protein efficiency ratio PER: wet weight gain/crude protein intake.

[0187] Retention (% of intake): 100×(FBW×final nutrients content in the carcass−IBW×initial nutrients content in the carcass)/nutrient intake.

[0188] 1.7. Statistical Analysis

[0189] The data are presented by the average of three repetitions±the standard deviation. The data were subjected to one-factor analysis of variance. Before ANOVA, the values expressed in % were subjected to an arcsine square root transformation. The statistical significance was tested at a probability level of 0.05. All the statistical tests were carried out using IBM SPSS V21 software.

2. Results

[0190] 2.1. Growth Performance

[0191] The data on the growth performances, feed conversion and protein efficiency of the rainbow trout fed with the experimental diets for 28, 60 and 90 days are reported in Tables 6-8 and FIG. 2. No deaths occurred during the test.

TABLE-US-00006 TABLE 6 Growth performances on day 28. Regime CTRL Y5 Y7.5 Y15 Y25 IBW (g)  5.0 ± 0.1  4.9 ± 0.1  5.0 ± 0.1  5.1 ± 0.1  5.1 ± 0.1 FBW (g) 16.1 ± 0.1.sup.a 16.2 ± 0.5.sup.a 16.2 ± 0.5.sup.a 17.9 ± 0.3.sup.b 17.6 ± 0.4.sup.b SGR, %/d 4.19 ± 0.12.sup.a 4.26 ± 0.13.sup.a 4.20 ± 0.07.sup.a 4.50 ± 0.07.sup.b 4.45 ± 0.06.sup.b FCR 0.87 ± 0.01.sup.b 0.87 ± 0.02.sup.b 0.87 ± 0.03.sup.b 0.81 ± 0.00.sup.a 0.81 ± 0.01.sup.a Feed intake, 3.27 ± 0.07 3.31 ± 0.09 3.28 ± 0.09 3.25 ± 0.03 3.22 ± 0.07 % BWM/d PER 2.55 ± 0.02.sup.a 2.56 ± 0.05.sup.a 2.55 ± 0.08.sup.a 2.66 ± 0.01.sup.ab 2.72 ± 0.05.sup.b The values are the averages ± the standard deviation (n = 3). The values within a row with different exponents differ significantly (P < 0.05).

[0192] After 28 days of experimental feeding (Table 6), the fish have more than tripled their initial body weight. The feed intake was high (3.22-3.31% BWM/day) and was not affected (P>0.05) by the increasing doses of beetle powder incorporated. This observation suggests that the beetle powder had no negative effect on palatability, and even that it could compensate for the total elimination of fishmeal without compromising the feed intake. The growth rate varied from 4.19 to 4.50%/day. In comparison with the CTRL treatment, while the Y5 and Y7.5 diets did not affect the FBW and the SGR, the Y15 and Y25 diets led to a significant increase (P<0.05) in FBW and SGR. The values of the feed conversion ratio vary between 0.81 and 0.87. In comparison with the CTRL, the inclusion of beetle powder at 5 and 7.5% (Y5 and Y7.5% diets) did not affect the FCR. However, the high levels of inclusion of beetle powder (Y15 and Y25 diets) led to a significant reduction in FCR (P<0.05). The protein efficiency ratio (PER) varied between 2.55 and 2.72. The fish fed with a Y25 diet showed a significant increase in PER, compared with those fed with the CTRL, Y5 and Y7.5 diets.

TABLE-US-00007 TABLE 7 Growth performances on day 60. Diet CTRL Y5 Y7.5 Y15 Y25 IBW (g)  5.0 ± 0.1  4.9 ± 0.1  5.0 ± 0.1  5.1 ± 0.1  5.1 ± 0.1 FBW (g) 30.3 ± 0.1.sup.a 31.6 ± 0.5.sup.a 34.9 ± 1.5.sup.b 37.2 ± 0.9.sup.c 42.9 ± 0.4.sup.d SGR, %/d 3.00 ± 0.04.sup.a 3.10 ± 0.04.sup.b 3.24 ± 0.04 .sup.c 3.31 ± 0.05.sup.c 3.57 ± 0.04.sup.d FCR 1.10 ± 0.03.sup.d 1.02 ± 0.03.sup.c 0.92 ± 0.01.sup.b 0.90 ± 0.02.sup.b 0.85 ± 0.02.sup.a PER 2.01 ± 0.06.sup.a 2.17 ± 0.06.sup.b 2.40 ± 0.02.sup.c 2.46 ± 0.06.sup.cd 2.56 ± 0.07.sup.d The values are the averages ± the standard deviation (n = 3). The values within a row with different exponents differ significantly (P < 0.05).

[0193] After 60 days of experimental feeding (Table 7), the fish undergoing the most effective treatment showed an increase of 8 times the initial body weight. The growth level varied from 3.00 to 3.57%/day. In comparison with the CTRL treatment, all the diets with the beetle powder showed a significant increase (P<0.05) in SGR. The FCR values varied between 0.85 and 1.10 and in comparison with the CTRL, the inclusion of the beetle powder at all the doses tested led to a significant reduction in FCR (P<0.05). The protein efficiency ratio (PER) varied between 2.01 and 2.56. The lowest PER value was found in the fish fed with a CTRL diet, while an improvement in PER was closely associated with increasing doses of the beetle powder

TABLE-US-00008 TABLE 8 Growth performances on day 90. Diet CTRL Y5 Y7.5 Y15 Y25 IBW (g)  5.0 ± 0.1  4.9 ± 0.1  5.0 ± 0.1  5.1 ± 0.1  5.1 ± 0.1 FBW (g) 42.9 ± 1.3.sup.a 45.2 ± 1.0.sup.b 49.0 ± 0.6.sup.c 51.0 ± 1.4.sup.c 55.9 ± 1.0.sup.d SGR, %/d 2.39 ± 0.06.sup.a 2.47 ± 0.02.sup.b 2.54 ± 0.03.sup.b 2.56 ± 0.05.sup.b 2.67 ± 0.04.sup.c FCR 0.93 ± 0.02.sup.b 0.83 ± 0.03.sup.a 0.80 ± 0.02.sup.a 0.79 ± 0.04.sup.a 0.79 ± 0.02.sup.a PER 2.38 ± 0.06.sup.a 2.68 ± 0.10.sup.b 2.76 ± 0.06.sup.b 2.80 ± 0.15.sup.b 2.74 ± 0.08.sup.b The values are the averages ± the standard deviation (n = 3). The values within a row with different exponents differ significantly (P < 0.05).

[0194] At the end of the test, 90 days of experimental feeding (Table 8), the fish undergoing the most effective treatment showed an increase of 11 times the initial body weight. In comparison with the CTRL fish, those fed with the diets rich in insects showed a significant increase in final body weight (P<0.05). This increase was dose-related, with a moderate increase for the Y5 diet, intermediate for Y7.5 and Y15, and highest for Y25. The specific growth rate (SGR) varied between 2.39 and 2.67%/day, with a minimal value found in the fish fed with a CTRL diet, while those fed with feeds containing beetle powder showed significantly higher SGR values (p<0.05). Independently of the level of incorporation, the beetle powder led to a significant reduction in FCR (P<0.05). In comparison with the CTRL treatment, all the diets comprising meals of insects led to a significant increase in the PER values (P<0.05).

[0195] 2.2. Composition of the Whole Body

[0196] The data on the composition of the whole body of the trout at the end of the test are presented in Table 9. The feeding treatments had no effect (P>0.05) on the moisture, protein, lipid, ash, phosphorus and energy levels of the whole fish.

TABLE-US-00009 TABLE 9 Composition of the whole body of the trout fed with the various feed treatments. Body composition CTRL Y5 Y7.5 Y15 Y25 Moisture, % 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 relative to the total weight of the fish. The values are the averages ± the standard deviation (n = 3). Initial fish: moisture 75.0%; protein 14.1%; fats 8.7%; ash 2.2%; phosphorus 0.4%, energy 6.7 kJ/g.

[0197] 2.3. Nutrient Retention

[0198] The nutrient and energy retention values (expressed in percentage of intake) are presented in Table 10. In comparison with the CTRL treatment, the fish fed with diets rich in beetle powder showed a significant increase in protein and energy retention (P<0.05). Similarly, the Y7.5, Y15 and Y25 diets showed a P retention significantly higher than the CTRL (P<0.05). Fat retention was not affected by the diets (P>0.05).

TABLE-US-00010 TABLE 10 Nutrient and energy retention in the trout fed with the various diets. Retention, % intake CTRL Y5 Y7.5 Y15 Y25 Protein 35.5 ± 2.5.sup.a 39.8 ± 0.7.sup.b 41.6 ± 0.4.sup.b 42.8 ± 2.2.sup.b 41.9 ± 2.2.sup.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.sup.a 32.7 ± 1.8.sup.ab 34.0 ± 0.7.sup.b 33.9 ± 1.7.sup.b 33.8 ± 1.1.sup.b Energy 42.0 ± 0.8.sup.a 45.4 ± 1.6.sup.b 47.1 ± 1.4.sup.b 47.8 ± 1.8.sup.b 48.0 ± 2.9.sup.b The values are the averages ± the standard deviation (n = 3). The values within a row with different exponents differ significantly (P < 0.05).

[0199] 2.4. Apparent Digestibility

[0200] The composition of the faecal matter collected from the trout fed with the various feed treatments is presented in Table 11.

TABLE-US-00011 TABLE 11 Composition of the faecal matter of the trout fed with the various diets. Composition of faecal matter CTRL Y5 Y7.5 Y15 Y25 Yttrium oxide, 1384 ± 39  1395 ± 94  1415 ± 61  1369 ± 62  1411 ± 43  (mg/kg) Protein, % DM* 19.63 ± 0.06 19.67 ± 0.24 19.76 ± 0.34 19.70 ± 0.38 19.20 ± 0.41 Fats, % DM*  4.37 ± 0.06  4.33 ± 0.19  4.28 ± 0.24  4.30 ± 0.06  4.20 ± 0.33 Phosphorus, %  2.64 ± 0.06  2.77 ± 0.08  2.65 ± 0.10  2.54 ± 0.15  2.62 ± 0.09 DM* Energy, kJ/g DM 23.24 ± 0.16 23.14 ± 0.40 23.47 ± 0.47 22.88 ± 0.16 23.09 ± 0.16 *% by weight relative to the total weight of dry matter in faecal matter. The values are the averages ± the standard deviation (n = 3).

[0201] The apparent digestibility coefficients (ADC %) for the different nutrients and energy are presented in Table 12. The increase in the doses of beetle powder incorporated had no significant effect (P>0.05) on the apparent digestibility of the dry matter, proteins, fat, phosphorus and energy.

TABLE-US-00012 TABLE 12 Apparent digestibility of the nutrients and energy in the trout. ADC, % CTRL Y5 Y7.5 Y15 Y25 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, 69.9 ± 1.4 68.3 ± 1.5 70.5 ± 2.4 71.4 ± 2.9 70.3 ± 1.8 % of intake Energy, 84.1 ± 0.4 84.3 ± 0.8 84.1 ± 1.0 84.2 ± 0.6 84.4 ± 0.6 % of intake The values are the averages ± the standard deviation (n = 3).

3. Conclusion

[0202] At the end of 90 days of experimental feeding, the overall growth performance can be considered as very satisfactory and in a higher range for the young rainbow trout, with SGR values for the total duration of the test varying between 2.4 and 2.7%/day. In the most effective treatments, the fish showed an increase of 11 times their initial body weight. The feed conversion rate among the treatments varied between 0.79 and 0.93, which suggests a good nutritional adequacy of the feeds and good feeding practices.

[0203] The experimental data generated in this example make it possible to affirm that: [0204] The incorporation of increasing doses of beetle powder (5, 7.5, 15 and 25%) with a concomitant reduction in fishmeal was progressively linked to a significant increase in the body weight of the fish. [0205] All the diets containing the beetle powder showed a significant improvement in SGR, FCR and PER. [0206] The increasing doses of beetle powder incorporated had no effect on the composition of the whole body of the trout. [0207] The increasing doses of beetle powder incorporated had no effect on the apparent digestibility of the dry matter, proteins, lipids, phosphorus and energy in the different experimental diets. [0208] The proteins, the phosphorus and energy retention were enhanced in trout fed with feeds comprising beetle powder.

[0209] In general, the beetle powder utilized in this example could effectively replace 100% of the fishmeal in the diet of juvenile rainbow trout with positive effects on FCR and overall growth performance.

EXAMPLE 5: METHODS WITH OR WITHOUT GRINDING PRIOR TO PRESSING

[0210] Method with Pressing Only

[0211] 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 brought to the boil beforehand. After 5 minutes, the beaker is removed from the water bath, the larvae are drained, then passed into a twin-screw-type press. A press cake is thus obtained.

[0212] Method with Grinding Followed by Pressing

[0213] 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 brought to the boil beforehand. 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 into a twin-screw-type press. A press cake is thus obtained.

[0214] Measurement of the Lipid Content

[0215] 2 g of sample is placed in a beaker to which 0.2 g of Na.sub.2SO.sub.4 and 15 mL of CHCl.sub.3/MeOH (2/1 v/v) are added. The mixture is placed under magnetic stirring for 20 minutes, then the solution is filtered, the residue is again placed in the beaker with 10 mL of CHCl.sub.3/MeOH (2/1 v/v). The mixture 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 by weight after extraction-evaporation relative to the initial weight of the sample (2 g).

[0216] Conclusion

[0217] The significance of grinding upstream of pressing was studied (FIG. 3). It is thus clearly apparent that the distribution of the lipids between the press cake and the press juice is much more effective, 12.9 versus 87.1 as opposed to 42.7 versus 57.3, when grinding was carried out beforehand.

EXAMPLE 6: ANALYSIS OF THE SIZE OF THE SOLUBLE PROTEINS OF THE BEETLE POWDER ACCORDING TO THE INVENTION

[0218] A sample of 100 mg of the beetle powder prepared in Example 1 was placed in 10 mL of NaCl phosphate buffer (pH 7.4, 0.137 mM). The sample was stirred for 1 minute (vortex), then centrifuged at 900 g for 1 min. After centrifugation, the sample was filtered through a 0.45 μm membrane. Analysis of the size of the soluble proteins was carried out using a steric exclusion chromatography system with a Nucleogel GFC-300 column. An NaCl phosphate buffer (pH 7.4, 0.137 mM) was used as eluent. The flow rate was 1.0 mL/min. Detection was carried out with a UV detector at 280 nm.

[0219] The results of the analysis are presented in FIG. 4 and summarized in Table 13 below.

TABLE-US-00013 TABLE 13 Distribution of the sizes of the soluble proteins contained in the beetle powder prepared in Example 1 Size of the proteins (kg/mol) Relative abundance (%)  6.5 to 12.4 74.4 12.4 to 29   20.5 29 to 66 5.1

[0220] The results show that approximately 74.4% of the soluble proteins present in the beetle powder according to the invention have a molar mass of less than 12,400 g/mol (or Da, Daltons).