Method for obtaining insects or insect larvae rich in vitamin d

Abstract

The present invention relates to a method for preparing insects or insect larvae rich in vitamin D, comprising raising insects or their larvae on an ergosterol-rich food substrate, for example fungi, in order to obtain ergosterol-rich insects or larvae and then exposing them to UV radiation. The present invention also relates to insects and insect larvae rich in vitamin D, and to compositions comprising same, in particular intended for human and / or animal food or for cosmetic use.

Description

[0001] DESCRIPTION

[0002] TITLE: PROCESS FOR OBTAINING INSECTS OR INSECT LARVAE RICH IN VITAMIN D

[0003] Field of invention

[0004] The present invention relates to compositions rich in vitamin D for human or animal consumption.

[0005] Technological background

[0006] Vitamin D is just as important for livestock as it is for humans. In addition to its essential role in calcium-phosphorus metabolism, it also plays a crucial role in modulating the immune system, growth, fertility, muscle function, and cardiac and mental health. A large portion of this vitamin is synthesized in the epidermis under the action of UV rays, while another portion is provided by diet. This is why animals raised indoors must be supplemented with vitamin D in their diet to compensate for the lack of endogenous synthesis.

[0007] Exposure of the skin to UV radiation results in the production of vitamin D by converting its precursor, 7-dehydrocholesterol, into cholecalciferol, known as vitamin D3. This conversion occurs through a non-enzymatic process called photolysis. It is also known that ergosterol (the equivalent of cholesterol in mushrooms) can be converted into ergocalciferol, known as vitamin D2, through exposure to UV-B radiation. Vitamins D2 and D3, collectively referred to as vitamin D, have virtually the same biological activities, and vitamin D2 synthesized by exposing mushrooms to UV light is used in many dietary supplements.

[0008] Industrialists are showing growing interest in using insects in animal feed and human nutrition, particularly due to their high nutritional protein content. In animal feed, the interest in replacing soy or fish protein with insect protein lies in the fact that insect farming would offer numerous environmental benefits, such as lower greenhouse gas production and lower water consumption. The black soldier fly (Hermetia lllucens) is a particularly interesting insect due to its ability to grow on a wide variety of organic substrates. It can therefore be used to recycle plant by-products or a wide variety of production waste.

[0009] Oonincx et al. (2018, Scientific Reports, 8: 10807,1-10) studied the synthesis of vitamins D2 and D3 in different insect species exposed to IIV-B radiation. The larvae of these insects were initially reared on conventional food substrates before being exposed to IIV-B radiation. A maximum concentration of 700 lU / kg of vitamin D2 (equivalent to 0.0175 mg / kg of vitamin D2) was obtained in Tenebrio molitor larvae after exposure to UV-B radiation for 64 hours. For the other insect species studied, the maximum concentration observed after UV-B exposure was approximately 275 lU / kg (equivalent to 0.0069 mg / kg) of vitamin D2 (for black soldier fly larvae).

[0010] There is still a need for sources of vitamin D for human and animal consumption.

[0011] Description of the invention

[0012] The inventors have demonstrated that it is possible to obtain insect larvae rich in vitamin D2, by raising said larvae on a food substrate rich in ergosterol, namely mushrooms, then exposing them to UV radiation. Furthermore, the vitamin D2 content is significantly increased if the larvae are fasted before exposure to UV radiation, this content being twice as high as that obtained with non-fasted larvae.

[0013] Larvae reared on mushrooms are thus able to bioaccumulate a very high concentration of ergosterol. Their subsequent exposure to UV radiation then allows the ergosterol they have bioaccumulated to be converted into vitamin D2.

[0014] The inventors have in fact shown that larvae exposed to UV radiation do not contain vitamin D2 if they have not previously consumed ergosterol.

[0015] Furthermore, exposure of larvae that have consumed mushrooms to UV radiation results in a vitamin D2 concentration in the larvae up to 14.5 times higher than the average vitamin D2 concentration observed when exposing mushrooms to UV radiation according to literature data.

[0016] Advantageously, the exposure of dead larvae, which have been previously raised on mushrooms, starved, and then killed by freezing, also leads to the production of larvae rich in vitamin D2.

[0017] The larvae obtained according to the invention constitute a source of vitamin D which can be at least 18 times more concentrated than the richest known food source of vitamin D, i.e. cod liver oil which contains 2.5 mg / kg of vitamin D. The results obtained with the black soldier fly larva can advantageously be obtained with larvae of other insect species deliberately enriched with ergosterol before exposure to UV radiation, for example larvae of Tenebrio molitor (mealworms) or common house fly. It is also possible to start from adult insects instead of insect larvae. Indeed, the process which allows ergosterol to be converted into vitamin D2 is a chemical process and does not require specific enzymes which might not be present in certain insect species.Specifically, when ergosterol is exposed to UV radiation, a photochemical reaction occurs, causing its conversion to previtamin D2 (7-dehydroergosterol). Previtamin D2 then undergoes spontaneous isomerization under the influence of heat to form ergocalciferol (vitamin D2). This also explains why exposing dead larvae that had bioaccumulated ergosterol to UV radiation also produces larvae rich in vitamin D2.

[0018] The production method according to the invention advantageously makes it possible to obtain insects or insect larvae rich in vitamin D, without raising them on a food substrate rich in this vitamin. Indeed, food substrates rich in vitamin D are in particular oily fish and it is not authorized, in Europe, to incorporate foods of animal origin into the diet of edible insects intended for animal or human nutrition.

[0019] Furthermore, the method according to the invention is relatively easy to implement, particularly at the industrial level, especially since exposure of mushrooms to UV radiation is already used to obtain mushrooms rich in this vitamin.

[0020] Advantageously, the mushroom-based food substrate can be based on mushroom waste (especially mushroom stalks). Indeed, similar results are obtained by rearing insect larvae on this mushroom waste versus whole mushrooms.

[0021] The present invention thus relates to a method for preparing insects or insect larvae rich in vitamin D, in particular comprising at least 5 mg / kg of vitamin D2 relative to the fresh weight of insect or insect larva, said method comprising the following steps: a) raising insects or insect larvae on a food substrate rich in ergosterol, in order to obtain insects or larvae rich in ergosterol, b) optionally, fasting the insects or larvae rich in ergosterol obtained in step a), in order to obtain insects or larvae rich in ergosterol and fasting, and c) exposing the insects or larvae rich in ergosterol obtained in step a) or b) to LIV radiation, in order to obtain insects or insect larvae rich in vitamin D.

[0022] Said insects or larvae are preferably selected from the group consisting of black soldier flies (Hermetia lllucens), Tenebrio molitor, Alphitobius diaperinus, Musca domestica, Acheta domesticus, Gryllodes sigillatus, steppe crickets (Gryllus assimilis) and larvae thereof.

[0023] Said ergosterol-rich food substrate preferably comprises mushrooms, algae and / or yeasts.

[0024] Insects or larvae exposed to UV radiation in step c) may be alive or dead.

[0025] Step c) comprises, for example, exposing the insects or larvae to UV radiation, preferably UV-B, for a period of at least 15 minutes.

[0026] Step c) comprises, for example, exposing the insects or larvae to UV radiation, preferably UV-B, of at least 2 J / cm 2 .

[0027] Another subject of the invention is a vitamin D-rich insect or insect larva comprising at least 5 mg / kg of vitamin D2. Said vitamin D-rich insect or insect larva is preferably obtained by exposing said insect or insect larva previously reared on an ergosterol-rich substrate to UV radiation.

[0028] Another subject of the invention is a composition comprising (i) insects or insect larvae rich in vitamin D as defined above or obtained by the process as defined above, (ii) an extract of said insects or said insect larvae or (iii) a derivative of said insects or said insect larvae.

[0029] Said composition is, for example, a food composition, a food supplement or a cosmetic composition.

[0030] Another subject of the invention relates to the use of insects or insect larvae rich in vitamin D as defined above or obtained by the process as defined above or of a composition as defined above, as a source of vitamin D.

[0031] Another subject of the invention relates to the use of a food substrate rich in ergosterol, to obtain insects or insect larvae rich in vitamin D after their exposure to UV radiation, preferably to UV-B radiation.

[0032] Another subject of the invention relates to the use of insects or insect larvae rich in vitamin D as defined above or obtained by the process as defined above, to obtain an extract of insects or insect larvae or a derivative of insects or insect larvae.

[0033] Insects and insect larvae rich in vitamin D

[0034] The present invention relates to an insect or insect larva rich in vitamin D, in particular rich in vitamin D2.

[0035] By "insect" we mean the adult insect.

[0036] Insect or insect larva as defined above is any insect or insect larva edible to humans and / or animals.

[0037] The insect can be, for example, a fly, a grasshopper, a cricket or an ant.

[0038] The insect or insect larva as defined above is for example of the genus Hermetia, Tenebrio, Alphitobius, Musca, Acheta, Gryllodes, Gryllus or Lasius.

[0039] The insect or insect larva as defined above is for example selected from the group consisting of a black soldier fly (Hermetia lllucens), mealworm (Tenebrio molitor), mealworm (Alphitobius diaperinus), a housefly (Musca domestica), a house cricket (Acheta domesticus), a zoo cricket (Gryllodes sigillatus), a steppe cricket (Gryllus assimilis) and a larva thereof.

[0040] The insect larva as defined above is preferably a black soldier fly larva {Hermetia lllucens) or a mealworm larva {Tenebrio molitor).

[0041] The expression "rich in vitamin D" or "rich in vitamin D2" here means an insect or an insect larva comprising at least 5 mg / kg of vitamin D2, preferably at least 7 mg / kg, more preferably at least 10 mg / kg, for example at least 20 mg / kg, at least 25 mg / kg, at least 30 mg / kg, at least 35 mg / kg or at least 40 mg / kg of vitamin D2.

[0042] Vitamin D concentrations (especially vitamin D2) in an insect or larva are always expressed here relative to the fresh weight of the insect or insect larva, for example in mg / kg fresh weight of the insect or insect larva. Fresh weight is the mass of the insect or insect larva without prior dehydration.

[0043] The insect or insect larva rich in vitamin D, in particular rich in vitamin D2, as defined above, thus comprises at least 5 mg / kg of vitamin D2, preferably at least 7 mg / kg, more preferably at least 10 mg / kg, for example at least 20 mg / kg, at least 25 mg / kg, at least 30 mg / kg, at least 35 mg / kg or at least 40 mg / kg of vitamin D2.

[0044] The vitamin D2 concentration can be measured by any suitable method well known to those skilled in the art, for example by hexane extraction followed by HPLC (high performance liquid chromatography). Vitamin D2 is then quantified at 265 nm. One such method is detailed in the example. The insect or larva is finely ground before hexane extraction.

[0045] The insect or insect larva rich in vitamin D, in particular rich in vitamin D2, as defined above, is preferably obtained by implementing the method as described below.

[0046] The insect or insect larva rich in vitamin D, in particular rich in vitamin D2, as defined above is obtained by exposing to UV radiation, preferably to UV-B radiation, said insect or said insect larva previously raised on a food substrate rich in ergosterol (for example comprising fungi, yeasts and / or algae), in particular by means of a lamp emitting UV radiation, said insect or said larva preferably having been fasted before said exposure, and said insect or said larva optionally being dead before said exposure.

[0047] Exposure to UV radiation, ergosterol-rich food substrate and fasting are particularly as described below.

[0048] The insect or insect larva rich in vitamin D, in particular vitamin D2, as defined above may be subjected to one or more subsequent processing steps, in particular to obtain a product usable in animal or human food.

[0049] For example, the insect or insect larva rich in vitamin D, in particular in vitamin D2, as defined above, can be frozen, processed to obtain an extract, for example to extract a protein fraction and / or an oil fraction and / or to obtain a derivative, for example to obtain a meal of insects or insect larvae rich in vitamin D, in particular in vitamin D2.

[0050] A meal of insects or insect larvae rich in vitamin D, in particular rich in vitamin D2, is for example obtained by a possible bleaching step (for example by immersing the insects or insect larvae briefly in hot water or by heating them with steam), a drying step (for example in a low-temperature oven, by air dehydration or by freeze-drying) and a grinding step (for example using a mill or grinder).

[0051] The protein fraction is obtained, for example, by solvent extraction, ultracentrifugation, filtration, and / or protein precipitation. The protein fraction is then preferably dried (for example, by spray drying, freeze-drying, or low-temperature drying), then preferably ground (for example, in a mill or grinder) and optionally sieved. The oil fraction is obtained, for example, by solvent-based extraction or by mechanical pressure. For solvent extraction, the insects or insect larvae are preferably ground and then mixed with a solvent, for example, hexane or ethanol. The solvent dissolves the fats and is separated from the resulting suspension. Then, the oil fraction is recovered by evaporating the solvent. For mechanical pressure extraction, the insects or insect larvae are, for example, subjected to mechanical pressure using a hydraulic press or a screw press.The oil thus extracted is then preferably filtered to remove impurities.

[0052] The insect or insect larva rich in vitamin D, particularly vitamin D2, as defined above, may be a whole insect or a whole larva.

[0053] The insect or insect larva rich in vitamin D, particularly vitamin D2, as defined above, may be dead or alive.

[0054] Advantageously, the insect or insect larva rich in vitamin D, in particular vitamin D2, as defined above, may further be rich in vitamin D3. For example, the insect or insect larva rich in vitamin D, in particular vitamin D2, as defined above, may comprise at least 3 mg / kg of vitamin D3 relative to the fresh weight of the insect or insect larva, in particular after exposure to UV radiation, in particular UV-B, of at least 9 J / cm 2, for example at least 9.88 J / cm 2 . When the insect or insect larva is fasted before exposure to UV radiation, in particular UV-B radiation, the vitamin D-rich insect or insect larva may further comprise at least 6 mg / kg of vitamin D3 relative to the fresh weight of the insect or insect larva, for example at least 8 mg / kg of vitamin D3 relative to the fresh weight of the insect or insect larva, in particular after exposure to UV radiation, in particular UV-B radiation, of at least 9 J / cm 2 , for example at least 9.88 J / cm 2 .

[0055] The insect or insect larva rich in vitamin D, in particular vitamin D2, as defined above, may be enriched with at least one other micronutrient (e.g. provitamin A, vitamin E and / or carotenoid). Such an insect or larva is for example obtained by rearing it on a food substrate rich in ergosterol and rich in this or these other vitamins or micronutrients.

[0056] Composition comprising insects or insect larvae rich in vitamin D, a fraction thereof or a derivative thereof

[0057] The present invention also relates to a composition comprising (i) insects rich in vitamin D, in particular in vitamin D2, as defined above and / or insect larvae rich in vitamin D, in particular in vitamin D2, as defined above, (ii) an extract of said insects or said insect larvae and / or (iii) a derivative of said insects or said insect larvae.

[0058] An extract of said insects or said insect larvae is for example the protein fraction or the oil fraction of said insects or said insect larvae.

[0059] The protein fraction and the oil fraction are for example obtained as described above.

[0060] The derivative of said insects or said insect larvae is for example a flour or an oil.

[0061] Insect or insect larvae meal is for example obtained as described above.

[0062] The composition as defined above preferably comprises at least 10% (i) of insects or insect larvae rich in vitamin D, in particular vitamin D2, (ii) of an extract of said insects or said insect larvae or (iii) of a derivative of said insects or said insect larvae, the percentage being a mass percentage.

[0063] The composition as defined above comprises for example at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% (i) of insects or insect larvae rich in vitamin D, in particular vitamin D2, (ii) of an extract of said insects or said insect larvae or (iii) of a derivative of said insects or said insect larvae, the percentage being a mass percentage.

[0064] The composition as defined above comprises for example at most 99%, at most 90%, at most 80%, at most 70%, at most 60%, at most 50% (i) of insects or insect larvae rich in vitamin, in particular in vitamin D2, (ii) of an extract of said insects or said insect larvae or (iii) of a derivative of said insects or said insect larvae, the percentage being a mass percentage.

[0065] For example, the composition as defined above comprises from 10% to 99%, for example from 20% to 99% or from 50% to 99% (i) of insects or insect larvae rich in vitamin D, in particular in vitamin D2, (ii) of an extract of said insects or of said insect larvae or (iii) of a derivative of said insects or of said insect larvae, the percentage being a mass percentage.

[0066] In the compositions as defined above, the mass percentage is therefore a mass percentage relative to the total mass of the composition.

[0067] In a particular embodiment, the composition as defined above consists of (i) insects or insect larvae rich in vitamin D, in particular vitamin D2, as defined above, (ii) an extract of said insects or said insect larvae or (iii) a derivative of said insects or said insect larvae.

[0068] The composition as defined above is preferably suitable for human and / or animal consumption or for cosmetic use.

[0069] The composition as defined above is for example a food composition, a food supplement or a cosmetic composition (in particular when the composition comprises an extract or a derivative of insects or insect larvae rich in vitamin D, in particular vitamin D2 in the form of an oil).

[0070] The composition as defined above, in particular the food supplement as defined above, may further comprise at least one vitamin other than vitamin D2 (e.g. vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B8, vitamin B9, vitamin B12, vitamin C, vitamin E and / or vitamin K) and / or at least one mineral (e.g. calcium, magnesium, potassium, chlorine and / or sulfur) and / or at least one trace element (e.g. iron, zinc, copper, iodine, chromium, fluorine and / or selenium) and / or at least one probiotic and / or at least one other ingredient, e.g. taurine. The food composition, in particular the food supplement, is for example in the form of a capsule.

[0071] The composition as defined above, in particular the food composition as defined above, is for example a meat substitute based on insect flour or insect larvae rich in vitamin D as defined above.

[0072] The composition as defined above, in particular the food composition as defined above, may further comprise at least one other food and / or at least one nutrient.

[0073] A "nutrient" means a food substance that can be directly assimilated, such as a protein, a carbohydrate, a lipid or a micronutrient (for example, a vitamin, a mineral, a trace element, an essential amino acid or a polyunsaturated fatty acid).

[0074] The micronutrient can be a phytomicronutrient, such as a polyphenol, carotenoid, phytosterol, phytoestrogen, or glucosinolate.

[0075] Breeding substrate rich in ergosterol

[0076] In the context of the present invention, insects or insect larvae are reared on a rearing substrate, also called a feed substrate, which is rich in ergosterol. The ergosterol-rich feed substrate comprises a source of ergosterol. The ergosterol source may be any source of ergosterol suitable as a feed for the insect species that will be used in animal and / or human food.

[0077] The source of ergosterol may for example include or consist of fungi, algae and / or yeasts.

[0078] The ergosterol-rich food substrate may, for example, include mushrooms, algae and / or yeasts.

[0079] A mushroom that can be used as a source of ergosterol in the food substrate is therefore preferably a mushroom that is edible for humans.

[0080] The mushroom as defined above is for example chosen from a mushroom of the genus Agaricus, Cantharellus, Craterellus, Gomphus, Hygrophoropsis, Pleurotus (also called oyster mushroom), Lentiluna, Flammulina, Grifola, Boletus or Morchella (also called morel).

[0081] The mushroom as defined above is for example Agaricus bisporus (also called button mushroom), Pleurotus ostreatus, Lentinula edodes (also called shiitake), Flammulina velutipes (also called enokitake), Grifola frondosa (also called maitake)

[0082] The mushroom as defined above is for example a chanterelle, such as Cantharellus cibarius, Cantharellus amethysteus, Cantharellus ferruginascens, Craterellus cinereus, Craterellus lutescens, Craterellus tubaeformis, Craterellus undulatus, Gomphus clavatus, Gomphus floccosus or Hygrophoropsis aurantiaca (also called "false chanterelle").

[0083] The mushroom as defined above is for example a morel, such as Morchella rotunda, Morchella esculenta, Morchella costata, Morchella conica or Mitrophora semilibera.

[0084] The mushroom is preferably Agaricus bisporus, a porcini, a chanterelle or a shiitake.

[0085] The mushroom as defined above can be used fresh or in dried form.

[0086] All or part of the mushroom can be used. For example, only the mushroom stems can be used.

[0087] The mushroom is preferably used cut into pieces.

[0088] The mushroom as defined above is preferably used fresh and cut into pieces. Yeast that can be used as a source of ergosterol in the food substrate is, for example, Saccharomyces cerevisae. The yeast can be used in fresh or dried form.

[0089] Algae that can be used as a source of ergosterol in food substrates include, for example, green algae or red algae. Examples of algae include Chlorella, Ulva, Porphyra, Laminaria, or Sargassum. Algae can be used fresh or dried.

[0090] The source of ergosterol as defined above (in particular fungus, algae and / or yeast) preferably comprises at least 1 mg of ergosterol / 100 g of said source of ergosterol, more preferably at least 2 mg of ergosterol / 100 g or at least 5 mg of ergosterol / 100 g, more preferably still at least 10 mg of ergosterol / 100 g or at least 20 mg of ergosterol / 100 g, for example at least 30 mg of ergosterol / 100 g, at least 40 mg of ergosterol / 100 g or at least 50 mg of ergosterol / 100 g.

[0091] The ergosterol concentration in an ergosterol source (e.g., fungus, algae, or yeast) may be measured by any suitable method well known to those skilled in the art. For example, the ergosterol concentration may be measured by double hexane extraction followed by HPLC (high-performance liquid chromatography). Ergosterol is then quantified at 282 nm. Such a method is detailed in the example.

[0092] Ergosterol is in particular quantified at 282 nm, after double extraction with hexane followed by separation, identification and quantification of this molecule by HPLC.

[0093] The ergosterol-rich food substrate as defined above may consist of a source of ergosterol if this allows good growth of insects or insect larvae.

[0094] The ergosterol-rich food substrate as defined above may further comprise at least one food for optimizing the growth of said insects or said larvae.

[0095] For example, a suitable feed for the growth of insects or insect larvae, particularly the black soldier fly or its larvae, includes wheat bran and / or soybean or sunflower meal.

[0096] In addition, the ergosterol-rich food substrate as defined above may comprise at least one other micronutrient, such as provitamin A, vitamin E or a carotenoid, e.g. lutein or lycopene. The ergosterol-rich food substrate as defined above comprises at least 5% of a source of ergosterol, preferably at least 10% of a source of ergosterol, more preferably at least 20%, at least 30%, at least 40% or at least 50% of a source of ergosterol, the percentage being a mass percentage and, preferably, said source of ergosterol comprising or consisting of fungi, yeasts and / or algae as defined above.

[0097] The ergosterol-rich food substrate may, for example, comprise at least 60%, at least 70%, at least 80% or at least 90% of an ergosterol source, the percentage being a mass percentage and, preferably, said ergosterol source comprising or consisting of fungi, yeasts and / or algae as defined above.

[0098] In the food substrate as defined above, the mass percentage is therefore relative to the total mass of the food substrate.

[0099] Process for obtaining insects or insect larvae rich in vitamin D

[0100] The present invention relates in particular to a process for preparing insects or insect larvae rich in vitamin D, in particular vitamin D2.

[0101] Insects and insect larvae rich in vitamin D, particularly vitamin D2, are in particular as defined below.

[0102] The method for preparing insects or insect larvae rich in vitamin D, in particular rich in vitamin D2, preferably comprises the following steps: a) rearing insects or insect larvae on a food substrate rich in ergosterol, in particular as defined above, in order to obtain insects or larvae rich in ergosterol, b) optionally, fasting the insects or larvae rich in ergosterol obtained in step a) to obtain insects or larvae rich in ergosterol and fasting, and c) exposing said insects or larvae rich in ergosterol obtained in step a) or b) to UV radiation, preferably to UV-B radiation, in order to obtain insects or insect larvae rich in vitamin D, in particular rich in vitamin D2.

[0103] Vitamin D-rich insects or insect larvae contain at least 5 mg / kg of vitamin D2 based on the fresh weight of the insect or insect larva.

[0104] Vitamin D2 is quantified in particular at 265 nm, after double extraction with hexane followed by separation, identification and quantification of this molecule (HPLC high performance liquid chromatography). Step a)

[0105] Step (a) comprises rearing insects or insect larvae on an ergosterol-rich food substrate, in order to obtain ergosterol-rich insects or insect larvae.

[0106] The insect or insect larva used in step (a) is any insect or insect larva which may be incorporated, in whole or in part, into human and / or animal feed.

[0107] The insect or insect larva is in particular as defined above.

[0108] As previously stated, by "insect" we mean the adult insect.

[0109] The insect or insect larva used in step a) is, for example, of the genus Hermetia, Tenebrio, Alphitobius, Musca, Acheta, Gryllodes, Gryllus or Lasius.

[0110] The insect or insect larva used in step a) is for example selected from the group consisting of a black soldier fly (Hermetia lllucens), mealworm (Tenebrio molitor), mealworm (Alphitobius diaperinus), housefly (Musca domestica), house cricket (Acheta domesticus), zoo cricket (Gryllodes sigillatus), steppe cricket (Gryllus assimilis) and a larva thereof.

[0111] The insect larva used in step a) is preferably a black soldier fly larva (Hermetia lllucens) or a mealworm larva (Tenebrio molitor).

[0112] Food substrate is the food given to insects or insect larvae, especially for their growth.

[0113] The ergosterol-rich food substrate therefore includes a source of ergosterol.

[0114] The feed substrate is in particular as defined above in the section “Livestock substrate comprising a source of ergosterol”.

[0115] The food substrate may consist of a source of ergosterol if this allows good growth of insects or larvae.

[0116] For example, the source of ergosterol comprises or consists of fungi, yeasts and / or algae, particularly as defined above.

[0117] The ergosterol-rich food substrate may further comprise at least one food for optimizing the growth of said insects or said larvae.

[0118] For example, a suitable feed for the growth of insects or insect larvae, particularly the black soldier fly or its larvae, includes wheat bran and / or soybean or sunflower meal.

[0119] In addition, the food substrate may include at least one other micronutrient, such as provitamin A, vitamin E or a carotenoid, e.g. lutein or lycopene. The insects or larvae obtained from the process will thus be both rich in vitamin D2 and rich in this or these micronutrients.

[0120] The ergosterol-rich food substrate comprises at least 5% of an ergosterol source, preferably at least 10% of an ergosterol source, more preferably at least 20%, at least 30%, at least 40% or at least 50% of an ergosterol source, the percentage being a mass percentage, and preferably, said ergosterol source comprising or consisting of fungi, yeasts and / or algae.

[0121] The ergosterol-rich food substrate may, for example, comprise at least 60%, at least 70%, at least 80% or at least 90% of an ergosterol source, the percentage being a mass percentage and, preferably, the ergosterol source comprising or consisting of fungi, yeasts and / or algae.

[0122] The ergosterol-rich food substrate preferably comprises at least 1 mg of ergosterol / 100 g of said food substrate, more preferably at least 2 mg of ergosterol / 100 g or at least 5 mg of ergosterol / 100 g, more preferably still at least 10 mg of ergosterol / 100 g or at least 20 mg of ergosterol / 100 g, for example at least 30 mg of ergosterol / 100 g, at least 40 mg of ergosterol / 100 g or at least 50 mg of ergosterol / 100 g.

[0123] Apart from the feeding substrate which is as described above, the other conditions for rearing the insects and insect larvae are the appropriate conditions well known to those skilled in the art for the insect species used.

[0124] When the process is intended to prepare insect larvae rich in vitamin D, step a) may be preceded by a step of seeding the food substrate with insect eggs which will produce insect larvae, preferably in a breeding box.

[0125] For example, to obtain between 20 g and 100 g of larvae, 150 to 200 mg of eggs can be inoculated onto, for example, 1 to 4 kg of food substrate.

[0126] Preferably, the eggs are placed above the substrate on an oviposition medium, allowing for example the eggs to hatch about 3 cm above the food substrate.

[0127] For example, the eggs are incubated at a temperature of 29 ± 1 °C and a relative humidity of 65 ± 5%. In step a), the larvae hatched from the eggs are reared, for example, at a temperature of 29 ± 1 °C and a relative humidity of 65 ± 5%, particularly under artificial light / dark cycles of 14:10.

[0128] Step a) for example has a duration of 12 to 14 days, preferably 13 days.

[0129] The feed substrate is preferably supplied in excess.

[0130] In the absence of step b), the insects or larvae may be frozen at the end of step a), for example at a temperature of -18°C or lower (for example at a temperature between -18°C and -80°C).

[0131] Step b) (optional)

[0132] Optional step b) includes starving the ergosterol-rich insects or larvae obtained in step a), to obtain ergosterol-rich and starved insects or larvae.

[0133] This fasting stage can advantageously allow for a higher concentration of vitamin D2 in insects or larvae after step c) of exposure to UV radiation. The hypothesis is that, during the fasting stage, insects or larvae continue to absorb ergosterol that is in the matrix of the food substrate in their digestive tract. Thus, when insects or larvae are exposed to UV radiation at the end of the fasting stage, a greater amount of ergosterol is present in the tissues of the insects or larvae. This ergosterol is theoretically much more accessible to UV radiation than the ergosterol from the matrix of the food substrate present in the digestive tract of insects or larvae.

[0134] Step b) is preferably carried out with insect larvae rich in ergosterol.

[0135] The larvae are preferably fasted for at least one day, more preferably at least two days.

[0136] During the starvation stage, the insect larvae are no longer in contact with the food substrate. For this purpose, the larvae can, for example, be separated from the food substrate using a sieve (e.g., 2 x 2 mm). They are then preferably washed with water, dried (e.g., with absorbent paper), and transferred to an empty box.

[0137] At the end of the starvation step, the larvae are preferably washed with water and dried (e.g. with absorbent paper). At the end of step b), the larvae may be frozen, for example at a temperature of -18°C or lower (e.g. at a temperature between -18°C and -80°C).

[0138] Step c)

[0139] Step c) comprises exposing the ergosterol-rich insects or larvae obtained in step a) or b) to UV radiation, in order to obtain vitamin D2-rich insects or insect larvae.

[0140] UV radiation can be UV-A, UV-B and / or UV-C radiation.

[0141] Preferably, the UV radiation is UV-B radiation. UV-B radiation is more efficient at converting ergosterol to vitamin D2.

[0142] Exposure to UV radiation can be achieved by exposure to a natural source of UV, such as sunlight, or by exposure to an artificial source of UV, such as a device that emits UV radiation.

[0143] Any device emitting UV radiation, in particular any lamp emitting UV radiation, pulsed or not, well known to those skilled in the art may be used.

[0144] For example, the UV-A lamp used emits in a wavelength range from about 315 to about 400 nanometers (nm).

[0145] For example, the UV-B lamp used emits in a wavelength range from about 280 to about 315 nanometers (nm).

[0146] For example, the UV-C lamp used emits in a wavelength range of approximately 100 to 280 nanometers (nm).

[0147] The power of the lamp emitting UV-B radiation is, for example, between 2.7 and 2.8 mW.cm2 / s.

[0148] The insects or larvae are preferably exposed to UV radiation, preferably UV-B, for a sufficient time, in particular depending on the dose of UV radiation administered, to obtain the desired quantity of vitamin D, in particular the desired quantity of vitamin D2 and, where appropriate, the desired quantity of vitamin D3.

[0149] The insects or larvae are preferably exposed to UV radiation, preferably UV-B, for a period of at least 15 minutes, more preferably at least 30 minutes, more preferably still at least 45 minutes or at least one hour. The insects or larvae are preferably exposed to UV radiation, preferably UV-B, for a period of less than 10 hours, preferably a period of less than 8 hours, more preferably less than 5 hours.

[0150] For example, the duration of exposure to UV radiation, preferably UV-B, may be from 15 minutes to 5 hours, for example from 15 minutes to 4 hours, 3 hours or 2 hours, preferably from 15 minutes to 1 hour.

[0151] Alternatively, insects or larvae can, for example, be exposed to UV radiation, preferably UV-B, for a period of 30 minutes to 1.5 hours, for example for a period of 45 minutes to 1 hour 15 minutes.

[0152] In step c), the insects or larvae are preferably exposed to UV radiation, preferably UV-B, of at least 2 J / cm 2 , for example at least 2.47 J / cm 2 , more preferably at least 4.94 J / cm 2 , more preferably still at least 7.41 J / cm 2 or at least 9.88 J / cm 2 . In step c), the insects or larvae are preferably exposed to UV radiation, preferably UV-B, of less than 50 J / cm 2 .

[0153] UV radiation of XJ / cm 2 means that the dose received (also called energy received) is XJ / cm 2 .

[0154] The dose received can be calculated by multiplying the received power by the exposure time. The received power can, for example, be measured using a radiometer.

[0155] The power received depends in particular on the power of the lamp used, the distance from the lamp and the exposure angle.

[0156] Insects or larvae are preferably exposed to UV-B radiation at a dose of 2 J / cm 2 at 50 J / cm 2 , preferably 2.47 J / cm 2 at 49.4 J / cm 2 , for example 2.47 J / cm 2 at 24.7 J / cm 2 or 2.47 J / cm 2 at 10 J / cm 2 .

[0157] The optimal dose can be determined by exposing insects or insect larvae to increasing doses of UV radiation and choosing, for example, a dose that achieves the highest concentration of vitamin D2 without significantly altering the organoleptic properties of the insects or insect larvae.

[0158] In an advantageous embodiment, the insects or larvae are exposed to UV-B radiation: for a duration of at least 15 minutes, more preferably at least 30 minutes, more preferably still at least 45 minutes or at least one hour,

[0159] - preferably for a duration of less than 2 hours, for example less than 1 hour 30 minutes, and - at a dose of 2 J / cm 2 at 50 J / cm 2 , preferably 2 J / cm 2 at 20 J / cm 2 , for example 2.4 J / cm 2 at 10 J / cm 2 .

[0160] Insects or larvae exposed to UV radiation are either alive or dead.

[0161] Dead insects or larvae have, for example, been frozen between steps a) and c) (in the absence of step b)) or between steps b) and c). Before exposure to UV radiation, the frozen larvae are preferably thawed and then dried (e.g. in an oven or by ventilation).

[0162] Surprisingly, the method according to the invention also makes it possible to enrich the insect or insect larva with vitamin D3. For example, the insect or insect larva obtained by the method according to the invention may also comprise at least 3 mg / kg of vitamin D3 relative to the fresh weight of the insect or insect larva, in particular after exposure to UV radiation, in particular UV-B, of at least 9 J / cm 2 , for example at least 9.88 J / cm 2 . When the method comprises step b) of fasting, the insect or insect larva obtained by the method may advantageously comprise at least 6 mg / kg of vitamin D3 relative to the fresh weight of the insect or insect larva, for example at least 8 mg / kg of vitamin D3 relative to the fresh weight of the insect or insect larva, in particular after exposure to UV radiation, in particular UV-B, of at least 9 J / cm 2 , for example at least 9.88 J / cm 2 .

[0163] Vitamin D3 is in particular quantified at 265 nm, after double extraction with hexane followed by separation, identification and quantification of this molecule by HPLC (high performance liquid chromatography).

[0164] Use of insects or insect larvae rich in vitamin D or of a composition comprising said insects or said larvae, an extract of said insects or said larvae or a derivative of said insects or said larvae

[0165] The present invention also relates to the use of insects or insect larvae rich in vitamin D, in particular in vitamin D2, or of a composition comprising (i) insects or insect larvae rich in vitamin D, in particular in vitamin D2, (ii) an extract of said insects or said insect larvae or (iii) a derivative of said insects or said insect larvae, as a source of vitamin D, in particular as a source of vitamin D2, in particular in humans or animals, for example for human or animal food or in cosmetics. The insects rich in vitamin D, in particular in vitamin D2, the insect larvae rich in vitamin D, in particular in vitamin D2, and the composition are in particular as defined above.

[0166] The insect larvae rich in vitamin D2 according to the invention can in fact contain more than 46 mg / kg of vitamin D2, i.e. approximately 18 times more than cod liver oil which is by far the richest source of vitamin D (with 2.5 mg / kg of vitamin D), whereas insect larvae in their natural state do not contain significant quantities of vitamin D (D2 or D3) allowing them to be qualified as a rich source of vitamin D.

[0167] The present invention also relates to the use of insects or insect larvae rich in vitamin D, in particular in vitamin D2, as defined above or obtained by the process as defined above, to obtain an extract of insects or insect larvae or a derivative of insects or insect larvae. The extract of insects or insect larvae and the derivative of insects or insect larvae are in particular as defined above.

[0168] Use of an ergosterol-rich feed substrate to obtain vitamin D-rich insects or insect larvae

[0169] The present invention also relates to the use of a food substrate rich in ergosterol, for example comprising fungi, yeasts and / or algae, to obtain insects or insect larvae rich in vitamin D, in particular in vitamin D2, after their exposure to UV radiation, preferably to UV-B radiation.

[0170] The feed substrate rich in ergosterol, insects rich in vitamin D, in particular vitamin D2, insect larvae rich in vitamin D, in particular vitamin D2 and exposure to UV radiation are in particular as defined above.

[0171] In particular, insects or larvae may be starved prior to exposure to UV radiation, particularly as described above.

[0172] Other characteristics and advantages of the invention will emerge more clearly from the examples which follow, given for illustrative and non-limiting purposes.

[0173] Figures

[0174] [Fig. 1] Figure 1 shows the concentration of ergosterol (A) and vitamin D2 (B), in mg / kg fresh weight, in mushrooms, exposed or not to UV, and in larvae reared on these mushrooms. The exposure of the mushrooms was carried out at a dose of 2.47 J / cm 2 . Bars represent the mean ±SEM (n=3). In Figures A and B, concentrations are not significantly different between larvae and the corresponding rearing substrate (A: p = 0.08 for unexposed mushrooms and p = 0.07 for exposed mushrooms; B: p = 0.52, two-way ANOVA, followed by pairwise comparison using two-tailed t-tests).

[0175] [Fig. 2] Figure 2 shows the vitamin D2 concentration in mushrooms exposed to UV-B radiation and in larvae reared on mushrooms not exposed to UV-B radiation. The larvae were exposed to UV-B radiation after consuming the mushrooms. Exposure of the mushrooms and larvae was carried out at a dose of 2.47 J / cm 2 Bars represent mean ±SEM (n=4). (*: p < 0.05; Student's t test).

[0176] [Fig. 3] Figure 3 shows the concentrations of vitamin D2 and vitamin D3, in mg / kg fresh weight, in live larvae reared on mushrooms not exposed to UV-B. After starvation, different groups of larvae were exposed to UV-B radiation for increasing durations: 15 minutes, 1 hour, 2 hours or 5 hours (i.e. in received energy equivalent 2.47, 9.88, 19.76 and 49.4 J / cm2). Bars represent the mean ±SEM (n = 4). Different lowercase letters indicate that the vitamin D2 means are significantly different from each other, different uppercase letters indicate that the vitamin D3 means are significantly different from each other: p < 0.05; two-way ANOVA followed by pairwise comparison using two-tailed t-tests.

[0177] [Fig. 4] Figure 4 shows the vitamin D2 concentration, in mg / kg fresh weight, in (i) live larvae before starvation, (ii) live larvae after starvation, and (iii) dead larvae after starvation and freezing. The different groups of larvae were reared on non-UV-exposed mushrooms and then exposed to UV-B radiation for 15 minutes or 5 hours (2.47 or 49.4 J / cm2). Bars represent the mean ± SEM (n=4). Bars with different letters indicate that the corresponding means are significantly different: p < 0.05; two-way ANOVA followed by pairwise comparison using two-tailed t-tests.

[0178] [Fig. 5] Figure 5 shows the vitamin D3 concentration, in mg / kg fresh weight, in (i) live larvae before starvation, (ii) live larvae after starvation, and (iii) dead larvae after starvation and freezing. The different groups of larvae were reared on non-UV-exposed mushrooms and then exposed to UV-B radiation for 15 minutes or 5 hours (2.47 or 49.4 J / cm2). Bars represent the mean ± SEM (n=4). Bars with different letters indicate that the corresponding means are significantly different p < 0.05; two-way ANOVA followed by pairwise comparison using two-tailed t-tests.

[0179] Example

[0180] Materials and Methods

[0181] (i) Substrate used for rearing black soldier fly larvae (BSF)

[0182] The ergosterol-rich food substrate consists of button mushrooms (Agaricus bisporus) or button mushroom stems.

[0183] (ii) Rearing of black soldier fly larvae (BSF)

[0184] The breeding protocol was implemented and carried out at the company BioMiMetiC (Boulbon, France). The objective was to obtain between 20 g and 100 g of larvae for each group. To achieve this, depending on their humidity, 1.5 kg to 4 kg of substrates were roughly cut for each group and 150 to 200 mg of eggs were inoculated into them.

[0185] The rearing procedure was as follows: first, after cutting the substrates and collecting the necessary mass of eggs, these were placed above each substrate on an oviposition rack allowing the eggs to hatch approximately 3 cm above the food substrates. The rearing boxes containing the substrates and eggs were placed in a rearing room set at a temperature of 29 ± 1 °C and a relative humidity of 65 ± 5%.

[0186] Eggs and hatched larvae were reared for 13 days under these same parameters, under artificial light / dark cycles of 14:10 (hours:hours). At the end of the rearing period, the larvae were separated from their food substrate by a sieve (2 x 2 mm). They were then washed in tap water, dried, and transferred to an empty plastic box. The larvae were fasted and therefore kept for 5 days in this box to empty their digestive tract of any remaining substrate. After this period, the larvae were washed again in tap water and dried using absorbent paper before being frozen at -80°C. The larvae were either exposed to UV-B alive, before or after fasting, or exposed to UV-B dead after fasting, freezing, and drying.

[0187] (iii) Exposure to UV-B

[0188] The lamp (DSP UV-B tube, OSRAM HNL, 24 W) (Osram Gmbh, Munich, Germany) was allowed to warm up for at least 30 minutes before exposing the larvae or fungi to UV-B. The power received at the location where the fungi or larvae were exposed was measured using a radiometer: 2.747 mW.cm2 / s. The energy received was then calculated by multiplying this power by the exposure time in seconds. For example, for a 15-minute UV-B exposure, the energy received was (2.747 / 1000) * (15*60) = 2.47 J / cm2.

[0189] (iv) Extraction and quantification of vitamins

[0190] First, 100 mg of finely ground LMSN or food substrate (mushroom) samples were homogenized in 500 µl of distilled water, then 500 µl of an ethanol solution, including retinol acetate at a concentration of 0.4 mg / l, was added, before performing a double extraction using hexane. The hexane phases were recovered after centrifugation at 1.257 xg for 10 minutes at 4°C and pooled, then evaporated under nitrogen to a dry film which was then solubilized in 1 mL of acetonitrile / methanol / water (60 / 38 / 2, v / v / v).

[0191] The resulting LMSN and food substrate samples were injected into the HPLC apparatus, including an UltiMate 3000 system (Thermo Fisher Scientific, San Jose, CA, USA). The injection volumes into the HPLC system were 15–50 μl, depending on the theoretical vitamin concentration of the sample. Ergocalciferol (vitamin D2) and cholecalciferol (vitamin D3) were quantified at 265 nm and ergosterol at 282 nm. All analyses were performed using a 250 x 4.6 nm ZORBAX Eclipse XDB-C18 5-μm column (Agilent Technologies, Montpellier, France). The column was maintained at a constant temperature (40°C). The analysis was performed with an acetonitrile / methanol / water (60 / 38 / 2, v / v / v) mobile phase at a flow rate of 1.5 mL / min. The HPLC system included an UltiMate 3000 system (Thermo Fisher Scientific, San Jose, CA, USA).Quantification was performed using Chromeleon software by comparing the area of ​​the peak to be quantified with the calibration range of the standard molecule.

[0192] Results

[0193] (i) Bioaccumulation of ergosterol and vitamin D2 in larvae

[0194] Figure 1 shows the concentration of ergosterol (A) and vitamin D2 (B) in mushrooms exposed or not to UV-B and in larvae that were fed with these mushrooms exposed or not to UV-B.

[0195] Regarding the ergosterol content of mushrooms, it is not significantly different whether they were exposed to UV-B or not. On the other hand, vitamin D2 is not detected in mushrooms not exposed to UV, whereas it is detected in mushrooms exposed to UV-B. Regarding larvae, their ergosterol content is equivalent to that of mushrooms, whether they were raised on mushrooms exposed to UV-B or not. Note that larvae raised on a substrate not containing ergosterol do not contain ergosterol (data not shown). Regarding vitamin D2, only larvae raised on mushrooms previously exposed to UV-B contain vitamin D2, which shows that the larvae have not synthesized vitamin D2. Finally, the vitamin D2 content of larvae raised on mushrooms previously exposed to UV-B is equivalent to that of these mushrooms.The larvae therefore bioaccumulate the vitamin D2 present in the substrate.

[0196] (ii) Synthesis of vitamin D2 in fungi or in larvae reared on fungi not exposed to UV-B,

[0197] Figure 2 shows vitamin D2 concentrations in mushrooms exposed to UV-B radiation and in larvae reared on mushrooms not exposed to UV-B, then exposed to UV-B. At the same dose of UV-B radiation exposure (2.47 J / cm 2 , i.e. an exposure time of 15 minutes with the lamp and distance used), the vitamin D2 concentration in the larvae was 14.07± 0.81 mg / kg, twice that obtained in the mushrooms. At this dose of UV-B, the mushrooms and larvae did not contain detectable vitamin D3.

[0198] (iii) Effect of duration of exposure to UV-B radiation on vitamin D concentration in larvae

[0199] Figure 3 shows the concentrations of vitamin D2 and vitamin D3 in larvae exposed to UV-B radiation for different durations. The concentration of vitamin D2 in larvae exposed to UV-B radiation for 15 minutes differed significantly from that obtained in larvae exposed for 1 hour, 2 hours and 5 hours (p < 0.001). However, this concentration was not significantly different between 1, 2 and 5 hours. Regarding vitamin D3, it was not detected in larvae exposed to UV-B radiation for 15 minutes. The concentration of vitamin D3 in larvae exposed to UV-B radiation for 1 hour was not significantly different from those obtained in larvae exposed for 2 hours or 5 hours (p > 0.2).

[0200] (iv) Effect of a fasting step, optionally followed by a freezing step, before exposure to UV-B radiation, on vitamin D synthesis by larvae Figures 4 and 5 show respectively the vitamin D2 and D3 concentrations in larvae exposed to UV-B radiation for 15 minutes or 5 hours, the larvae being at the time of exposure (i) live larvae before fasting, (ii) live larvae after fasting or (iii) dead larvae after fasting and freezing. Note that all these larvae were previously reared on mushrooms.

[0201] Table 1 below shows the vitamin D2 concentration obtained in live larvae (before or after fasting) and dead larvae (after fasting), after different durations of exposure to UV-B radiation.

[0202] The results obtained after 15 minutes and 5 hours of exposure are also shown in Figure 4 and those obtained with live larvae after fasting are shown in Figure 3.

[0203] Exposure of between 15 minutes and 5 hours produces larvae rich in vitamin D2. It seems unnecessary to extend exposure beyond 1 hour, since the vitamin D2 concentration after more than 1 hour of exposure is not higher than that obtained after 1 hour of exposure (see figures 3 and 4).

[0204] Fasting larvae before exposure to UV radiation significantly increased vitamin D2 concentrations at 15 minutes compared to larvae exposed before fasting (p = 0.001) (see Figure 4). Freezing larvae after fasting decreased vitamin D2 concentrations obtained at 15 minutes and 5 hours (see Figure 4). However, during a 15-minute exposure, the vitamin D2 concentrations of dead larvae were not significantly different from those obtained in live larvae (p > 0.055). During a 5-hour exposure, vitamin D2 concentrations in the three conditions tested were not significantly different.

[0205] However, the vitamin D2 concentration obtained after fasting and freezing remains very interesting for the use of larvae as a source of vitamin D2 in human and animal food, because they can still be considered a rich source of vitamin D.

[0206] [Table 1] Vitamin D2 concentration (mg / kg fresh weight) of black soldier fly larvae after different durations of exposure to UV radiation

[0207] ND: Not determined

[0208] For comparison, exposing white button mushrooms (the substrate used for larval food) to UV radiation for 15 minutes yields 7.5 ± 1.7 mg / kg of vitamin D2, equivalent to 750 pg / 100 g fresh weight. Similar experiments in the literature have yielded 320 pg / 100 g fresh weight. The difference may be explained by the fact that in the present experiment, both sides of the mushrooms were exposed to UV radiation, which is apparently not the case in the experiments in the literature.

[0209] Finally, vitamin D3 was not detected under any of the conditions tested during a 15-minute exposure to UV-B radiation (see Figure 5). During a 5-hour exposure to UV-B radiation, vitamin D3 concentrations in live larvae before fasting and in dead larvae (after fasting and freezing) were not significantly different (p = 0.57), but these concentrations were significantly lower than those obtained in live larvae after fasting (p < 0.01).

Claims

CLAIMS 1. A method for preparing vitamin D-rich insects or insect larvae, said method comprising the following steps: a) rearing insects or insect larvae on an ergosterol-rich food substrate, in order to obtain ergosterol-rich insects or larvae, b) optionally, fasting the ergosterol-rich insects or larvae obtained in step a), in order to obtain ergosterol-rich and fasted insects or larvae, and c) exposing the ergosterol-rich insects or larvae obtained in step a) or b) to LIV radiation, in order to obtain vitamin D-rich insects or insect larvae.

2. Method according to claim 1, characterized in that said insects or said larvae are selected from the group consisting of black soldier flies, Tenebrio molitor, Alphitobius diaperinus, Musca domestica, Acheta domesticus, Gryllodes sigillatus, steppe crickets (Gryllus assimilis) and larvae thereof.

3. Method according to claim 1 or 2, characterized in that the food substrate rich in ergosterol comprises fungi, algae and / or yeasts.

4. Method according to one of claims 1 to 3, characterized in that the insects or larvae exposed to UV radiation in step c) are alive or dead.

5. Method according to one of claims 1 to 4, characterized in that step c) comprises exposing the insects or larvae to UV radiation for a period of at least 15 minutes.

6. Method according to one of claims 1 to 5, characterized in that step c) comprises exposing the insects or larvae to UV radiation of at least 2 J / cm 2 .

7. Insect or insect larva rich in vitamin D, characterized in that said insect or said larva comprises at least 5 mg / kg of vitamin D2 relative to the fresh weight of insect or insect larva and is obtained by exposing said insect or said larva previously raised on a substrate rich in ergosterol to UV radiation.

8. Composition comprising (i) insects or insect larvae rich in vitamin D according to claim 7 or obtained by the process according to one of claims 1 to 6, (ii) an extract of said insects or said insect larvae or (iii) a derivative of said insects or said insect larvae.

9. Composition according to claim 8, characterized in that it is a food composition, a food supplement or a cosmetic composition.

10. Use of insects or insect larvae rich in vitamin D according to claim 7 or obtained by the process according to one of claims 1 to 6 or of a composition according to claim 8 or 9, as a source of vitamin D.

11. Use of a food substrate rich in ergosterol, to obtain insects or insect larvae rich in vitamin D after their exposure to UV radiation.

12. Use of insects or insect larvae rich in vitamin D according to claim 7 or obtained by the process according to one of claims 1 to 6 to obtain an extract of insects or insect larvae or a derivative of insects or insect larvae.

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