New CNSL-derived composition and use thereof for specific application in food or pharmaceuticals

Abstract

The invention relates to a new composition of an extract of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated anacardic acid, and to the use of same for specific application in food, neutraceuticals or pharmaceuticals.

Description

[0001] New composition derived from CNSL and its use for a specific application for food or pharmaceutical purposes

[0002] The invention relates to a new composition derived from a cashew nut shell extract (CNSL) and its use for a specific application for food, nutraceutical or pharmaceutical purposes.

[0003] CONTEXT OF THE INVENTION

[0004] Cashew nut shell liquid (CNSL) is a natural extract derived from cashew nut shells. The main components of raw (natural) CNSL are phenolic compounds: anacardic acid, cardol, and cardanol. Methyl cardol is also present, but in trace amounts (<5%).

[0005] Each of these compounds is itself a mixture of products, comprising an alkyl or alkenyl chain, said alkenyl chain having 1, 2 or 3 double bonds (Diagram 1): anacardic acid cardol methyl cardol cardanol

[0006] 65% 15 to 20% trace 10%

[0007] Scheme 1 - Chemical structures of the main constituents of natural CNSL.

[0008] CNSL can be classified into 2 types, depending on the extraction method used:

[0009] • Natural CNSL, an extract obtained by extraction using a low boiling point solvent or obtained mechanically without heating such as by mechanical extraction under pressure,

[0010] • Technical CNSL, an extract obtained by hot processes, in particular by high-temperature roasting, e.g. >200°C. Technical CNSL contains a reduced amount of anacardic acid compared to natural CNSL, due to partial decarboxylation of this acid during heating or roasting. In some cases, decarboxylation is even complete, and the CNSL no longer contains anacardic acid.

[0011] By "crude, undecarboxylated CNSL" or "natural CNSL" we mean CNSL that has not undergone, or has only partially undergone, decarboxylation of anacardic acid. Crude, undecarboxylated CNSL is therefore characterized by the presence of anacardic acid as the majority species within the mixture.

[0012] To date, the valorization of CNSL oil is a promising avenue given the potential presented by the components that constitute it.

[0013] The use of natural or technical CNSL and that of its phenolic constituents with unsaturated C15 alkyl chain is known in the prior art as a food supplement or as a therapeutic agent, in particular to reduce methanogenesis in ruminants and to treat digestive pathological conditions such as abdominal tympanism or coccidiosis. However, the use of natural CNSL also has negative effects on digestive fermentation, as well as astringent and irritant effects, limiting its use.

[0014] The dose available in the literature for food use is between 50 and 500 ppm, or between 50 and 500 g of active ingredient per ton of feed ingested. CNSL registrations according to US legislation such as AAFCO (Association of American Feed Control Officials) mention and recommend an inclusion level of 500 to 600 ppm as a maximum in feed.

[0015] There is a need to promote CNSL and its constituents as natural and ecological alternatives to petrochemical products.

[0016] There is a need to valorize CNSL compounds, derived from agricultural production waste, in the food sector.

[0017] There is a need for dietary supplements or pharmaceutical compositions to treat digestive pathological conditions such as abdominal tympanism or coccidiosis.

[0018] There is a need for a food supplement to reduce methanogenesis, particularly in ruminants, which is the source of greenhouse gases.

[0019] There is a need for a feed supplement to improve the digestibility of organic matter or nutrients to promote growth and weight gain in animals.

[0020] One of the aims of the invention is to provide a new composition of natural origin or derived from plant and natural sources. Another aim of the invention is to provide a new food or therapeutic active ingredient.

[0021] Another aim of the invention is the non-therapeutic use of a new composition to reduce methanogenesis and / or promote digestive fermentation and / or optimize the composition of the intestinal flora (the microbiota).

[0022] Another object of the invention is to provide a food or a food supplement.

[0023] Another aim of the invention is a food composition making it possible to prevent or treat a digestive pathological condition such as abdominal tympanism or coccidiosis.

[0024] Another object of the invention is a non-therapeutic method for reducing methanogenesis.

[0025] A first subject of the present invention relates to the non-therapeutic use of a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV): or the non-therapeutic use of hydrogenated anacardic acid of formula (I) to decrease methanogenesis and / or to improve digestive fermentation in animals.

[0026] Advantageously, in said non-therapeutic use as defined above, methods of treating the human or animal body by therapy are excluded.

[0027] Advantageously in a particular embodiment, the animals are non-human, namely humans are excluded.

[0028] “Cashew nut shell extract” means a solid or liquid fraction obtained from cashew nut shells by a physical, thermal or chemical extraction and / or transformation process.

[0029] “Cashew nut shell liquid” or “CNSL” means a liquid extract obtained from cashew nut shells by a physical, thermal or chemical extraction and / or transformation process.

[0030] Thus, “natural CNSL” is advantageously obtained from an extraction process which does not involve or only partially involves the decarboxylation of anacardic acid, such as extraction by a solvent or by mechanical press.

[0031] "Technical CNSL" is produced by extraction with or followed by thermal processing resulting in partial or even total decarboxylation of anacardic acid. Anacardic acid then no longer represents the majority constituent. Advantageously, "technical CNSL" can be prepared from natural CNSL by thermal processing resulting in partial or total decarboxylation of anacardic acid.

[0032] By "hydrogenated cashew nut shell liquid" or "hydrogenated CNSL" is meant a natural or technical cashew nut shell liquid in which the phenolic constituents such as anacardic acid, cardol, methyl cardol and cardanol are predominantly present with a single saturated C15 alkyl chain.

[0033] Thus, hydrogenated cashew nut shell liquid (hydrogenated CNSL) is free from or contains less than 5%, in particular less than or equal to 2%, by total weight of unsaturated C15 alkyl chain anacardic acid, unsaturated C15 alkyl chain cardol, unsaturated C15 alkyl chain methyl cardol and unsaturated C15 alkyl chain cardanol.

[0034] Advantageously, the hydrogenated CNSL contains at least or more than 95% by weight of the phenolic species, namely by total weight of anacardic acid with a saturated C15 alkyl chain of formula (I), cardol with a saturated C15 alkyl chain of formula (II), methyl cardol with a saturated C15 alkyl chain of formula (III) and cardanol with a saturated C15 alkyl chain of formula (IV). Advantageously, the “hydrogenated CNSL”, in particular the “hydrogenated natural CNSL” and the “hydrogenated technical CNSL”, are in the form of a solid, in particular in the form of a powder.

[0035] Natural or technical CNSL is mainly in liquid form. Hydrogenation of the C15 alkyl chains of its phenolic constituents transforms it into hydrogenated CNSL, which is in solid form.

[0036] “Hydrogenated cashew nut shell extract” means a solid or liquid fraction obtained from cashew nut shells by a process of extracting cashew nut shells, followed by a chemical transformation by hydrogenation of the alkene functions of the alkyl chains of the constituents, so that the phenolic constituents such as anacardic acid, cardol, methyl cardol and cardanol have a saturated alkyl chain.

[0037] The extract can be in liquid or solid form only or in the form of a mixture of liquid and solid which can have a pasty texture.

[0038] For example, CNSL, whether natural or technical, is in a liquid state under normal temperature (25-30°C) and pressure (1 atmosphere) conditions. CNSL, whether natural or technical, is hydrogenated, i.e. having undergone hydrogenation to saturate the C15 alkyl chains of the phenolic constituents, at a rate of 95 to 100% of these C15 alkyl chains, is in a solid state under the same temperature and pressure conditions. CNSL having partially undergone hydrogenation of the C15 alkyl chains on a portion of the phenolic constituents is in a mixture of liquid and solid.

[0039] Hydrogenated CNSL is advantageously obtained by hydrogenation of the alkene functions present in the C15 alkyl chain of the constituents of natural or technical CNSL.

[0040] Advantageously, the “hydrogenated natural CNSL” is obtained by hydrogenation of the “natural CNSL”, in particular by hydrogenation at a temperature lower than the decarboxylation temperature of anacardic acid.

[0041] Advantageously, the “hydrogenated technical CNSL” is obtained by hydrogenation of the “technical CNSL”. Preferably, said “technical CNSL” is obtained from the “natural CNSL” by heat treatment to decarboxylate the anacardic acid.

[0042] "Methanogenesis" refers to the production of methane in the digestive system by microorganisms present in the animal or human microbiota, particularly in the rumen of ruminant animals. "Improving digestive fermentation" refers to improving the digestibility of ingested organic matter or nutrients and / or optimizing the composition of the microbiota, possibly with an increase in propionate production.

[0043] Indeed, ruminal digestive fermentation is defined by the fermentation of microorganisms that constitute the ruminal flora. This fermentation allows the digestion of fibrous materials (grasses, hay, etc.) and transforms them into bacterial proteins (approximately 2 kg / day) and others, which will feed the animal. If digestive fermentation is inhibited, the digestibility of organic matter is also inhibited and the production of these proteins and others will be reduced and the animal will produce less meat or milk.

[0044] More specifically, "digestive improvement in ruminants" means the optimization of ruminal fermentation in order to better digest the food ingested by the ruminant, namely the cow, ox, goat, sheep or ewe.

[0045] More specifically, "digestive improvement in monogastrics" means an improvement in the composition of the intestinal flora (the microbiota).

[0046] Thus, digestive fermentation is directly linked to the digestibility of organic matter. For the purposes of the present invention, the improvement in the digestibility of organic matter is due to the improvement of digestive fermentation.

[0047] The term "digestibility of organic matter" refers to the degree to which organic matter is digested by an animal.

[0048] The digestibility of organic matter and the methods of evaluation are known to those skilled in the art. As a non-limiting example, the digestibility of organic matter can be evaluated and calculated as described in Van Gastelen et al. (J. Dairy Sci. 104:4174-4191, 2021).

[0049] It is known that one of the problems associated with the use of natural CNSL and unsaturated anacardic acids, at the doses tested, in ruminants, is the reduction of ruminal fermentation because natural CNSL inhibits, through its antibacterial and antifungal properties, the fermentation of microorganisms that constitute the ruminal flora.

[0050] The inventors have surprisingly found that hydrogenated natural CNSL and hydrogenated anacardic acid lead to a reduction in methanogenesis without any negative effect on digestive fermentation, limiting the side effects of natural CNSL. Indeed, as indicated in the in vitro study of the examples, hydrogenated natural CNSL, comprising mainly hydrogenated anacardic acid, allows a reduction in gas production, in particular methanogenesis, a shift towards the production of propionate and valerate to the detriment of acetate and butyrate, with minimal and attenuated side effects on fermentation, by promoting the digestibility of organic matter and the maintenance of digestive pH.

[0051] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, of a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV): wherein said composition comprises:

[0052] - hydrogenated anacardic acid from 50.0 to 85.0% by total weight of the composition,

[0053] - hydrogenated cardol from 10.0 to 25.0% by total weight of the composition

[0054] - hydrogenated methyl cardol from 0.0 to 5.0% by total weight of the composition, and

[0055] - hydrogenated cardanol from 2.0 to 15.0% by total weight of the composition, or the non-therapeutic use of hydrogenated anacardic acid of formula (I) to reduce methanogenesis and / or to improve digestive fermentation in animals.

[0056] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, in which said composition is derived from an extract of cashew nut shells obtained by extraction using a solvent or mechanically without heating.

[0057] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, in which said composition is derived from an extract of cashew nut shells obtained by extraction using a solvent or mechanically without heating, followed by hydrogenation.

[0058] Advantageously, said composition is in solid form, in particular in powder form.

[0059] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, of a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or of a hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV):

[0060] or the non-therapeutic use of hydrogenated anacardic acid of formula (I) to decrease methanogenesis and possibly to improve digestive fermentation in animals.

[0061] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, of a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV):

[0062] or the non-therapeutic use of hydrogenated anacardic acid of formula (I) for improving digestive fermentation in animals. According to a particular embodiment, the invention relates to the use as defined above, of a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV):

[0063] to reduce methanogenesis and / or to improve digestive fermentation in animals.

[0064] According to a particular embodiment, the invention relates to the use as defined above, of a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising at least 95% by total weight of the combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV), for reducing methanogenesis and / or for improving digestive fermentation in animals.

[0065] Advantageously, said composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract is free from or contains an amount of less than 5%, in particular less than or equal to 2%, by weight of anacardic acid with unsaturated alkyl chain, cardol with unsaturated alkyl chain, methyl cardol with unsaturated alkyl chain and cardanol with unsaturated alkyl chain. According to a particular embodiment, the invention relates to the use as defined above, of a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract, consisting of the combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV), for reducing methanogenesis and / or for improving digestive fermentation in animals.

[0066] According to a particular embodiment, the invention relates to the non-therapeutic use of hydrogenated anacardic acid of formula (I), as defined above, for reducing methanogenesis and possibly improving digestive fermentation in animals.

[0067] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, for reducing methanogenesis and for improving digestive fermentation in animals.

[0068] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, for increasing the production of propionate.

[0069] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, for increasing the production of valerate.

[0070] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, for increasing the production of propionate and valerate.

[0071] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, for maintaining the rate of production of volatile fatty acids (VFA) or limiting its reduction.

[0072] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, to maintain the rate of production of acetate and / or butyrate or to limit their reduction.

[0073] According to a particular embodiment, the invention relates to the non-therapeutic use as defined above, for maintaining the pH or limiting its increase in the stomach or rumen of the animal.

[0074] According to a particular embodiment, the invention relates to the use as defined above, in which said composition of hydrogenated cashew nut shell liquid or hydrogenated cashew nut extract comprises:

[0075] - hydrogenated anacardic acid from 50.0 to 85.0% by total weight of the composition, - hydrogenated cardol from 10.0 to 25.0% by total weight of the composition

[0076] - hydrogenated methyl cardol from 0.0 to 5.0% by total weight of the composition, and

[0077] - hydrogenated cardanol from 2.0 to 15.0% by total weight of the composition.

[0078] The range “50.0 to 85.0%” includes the following ranges: 50.0 to 55.0%; 55.0 to 60.0%; 60.0 to 65.0%; 65.0 to 70.0%; 70.0 to 75.0%; 75.0 to 80.0%; 80.0 to 85.0%.

[0079] The range “from 10.0 to 25.0%” includes the following ranges: from 10.0 to 11.0%; from 11.0 to 12.0%; from 12.0 to 13.0%; from 13.0 to 14.0%; from 14.0 to 15.0%; from 15.0 to 16.0%; from 16.0 to 17.0%; from 17.0 to 18.0%; from 18.0 to 19.0%; from 19.0 to 20.0%; from 20.0 to 21.0%; from 21.0 to 22.0%; from 22.0 to 23.0%; from 23.0 to 24.0%; from 24.0 to 25.0%.

[0080] The range “0.0 to 5.0%” includes the following ranges: 0.0 to 1.0%; 1.0 to 2.0%; 2.0 to 3.0%; 3.0 to 4.0%; 4.0 to 5.0%.

[0081] The range “from 2.0 to 15.0%” includes the following ranges: from 2.0 to 3.0%; from 3.0 to 4.0%; from 4.0 to 5.0%; from 5.0 to 6.0%; from 6.0 to 7.0%; from 7.0 to 8.0%; from 8.0 to 9.0%; from 9.0 to 10.0%; from 10.0 to 11.0%; from 11.0 to 12.0%; from 12.0 to 13.0%; from 13.0 to 14.0%; from 14.0 to 15.0%.

[0082] According to a particular embodiment, the invention relates to the use as defined above, wherein said composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract or hydrogenated anacardic acid is used at a dose of 50 to 600 ppm, preferably 50 to 500 ppm.

[0083] The "50 to 600 ppm" range includes the following ranges: 50 to 60 ppm, 60 to 70 ppm, 70 to 80 ppm, 80 to 90 ppm, 90 to 100 ppm, 100 to 110 ppm, 110 to 120 ppm, 120 to 130 ppm, 130 to 140 ppm, 140 to 150 ppm, 150 to 160 ppm, 160 to 170 ppm, 170 to 180 ppm, 180 to 190 ppm, 190 to 200 ppm, 200 to 210 ppm, 210 to 220 ppm, 220 to 230 ppm, 230 to 240 ppm, 240 to 250 ppm, 250 to 260 ppm, 260 to 270 ppm, 270 to 280 ppm, 280 to 290 ppm, 290 to 300 ppm, 300 to 310 ppm, 310 to 320 ppm, 320 to 330 ppm, 330 to 340 ppm, 340 to 350 ppm, 350 to 360 ppm, 360 to 370 ppm, 370 to 380 ppm, 380 to 390 ppm, 390 to 400 ppm, 400 to 410 ppm, 410 to 420 ppm, 420 to 430 ppm, 430 to 440 ppm, 440 to 450 ppm, 450 to 460 ppm, 460 to 470 ppm, 470 to 480 ppm, 480 to 490 ppm, 490 to 500 ppm, 500 to 510 ppm, 510 to 520 ppm, 520 to 530 ppm, 530 to 540 ppm, 540 to 550 ppm, 550 to 560 ppm, 560 to 570 ppm,from 570 to 580 ppm, from 580 to 590 ppm, from 590 to 600 ppm.,

[0084] The unit "ppm", namely parts per million, is a mass fraction corresponding to 1 mg / kg.

[0085] According to a particular embodiment, the invention relates to the use as defined above, wherein said composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract is used at a dose of 50 to 600 ppm, preferably 50 to 500 ppm.

[0086] According to a particular embodiment, the invention relates to the use as defined above, in which the hydrogenated anacardic acid is used at a dose of 50 to 600 ppm, preferably 50 to 500 ppm.

[0087] According to a particular embodiment, the invention relates to the use as defined above, for reducing methanogenesis and possibly improving digestive fermentation in the rumen of ruminants and / or possibly for increasing the production of propionate, in particular of cattle.

[0088] According to a particular embodiment, the invention relates to the use as defined above, for reducing methanogenesis and possibly improving digestive fermentation in the rumen of ruminants, in particular cattle.

[0089] According to a particular embodiment, the invention relates to the use as defined above, for reducing methanogenesis and possibly for increasing the production of propionate, in particular in cattle.

[0090] According to a particular embodiment, the invention relates to the use as defined above, for reducing methanogenesis and possibly improving digestive fermentation in the rumen of cattle.

[0091] According to a particular embodiment, the invention relates to the use as defined above, for improving the digestive fermentation of animals.

[0092] According to a particular embodiment, the invention relates to the use as defined above, for improving digestive fermentation in ruminants, in particular cattle, sheep and goats, preferably cows, beef, goats, sheep and ewes. According to a particular embodiment, the invention relates to the use as defined above, for improving digestive fermentation in monogastric animals, in particular by influencing the composition of the intestinal flora. Another subject of the present invention relates to a composition of cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV) following: or a composition comprising or consisting of hydrogenated anacardic acid of formula

[0093] (I) for its use in reducing methanogenesis and / or in improving digestive fermentation in animals. According to a particular embodiment, the invention relates to the composition as defined above for the uses described above. Another subject of the present invention relates to the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV):

[0094] According to a particular embodiment, the invention relates to the association as defined above, in which

[0095] - hydrogenated anacardic acid represents from 50.0 to 85.0% by total weight of the association,

[0096] - hydrogenated cardol represents from 10.0 to 25.0% by total weight of the association,

[0097] - hydrogenated methyl cardol represents from 0.0 to 5.0% by total weight of the association, and

[0098] - hydrogenated cardanol represents from 2.0 to 15.0% by total weight of the association.

[0099] According to a particular embodiment, the invention relates to the association as defined above, in which said association is derived from a hydrogenated cashew nut shell extract, in particular hydrogenated CNSL, preferably hydrogenated natural CNSL. Advantageously, said association as defined above is derived from a natural CNSL having undergone hydrogenation of the C15 alkyl chains of its constituents.

[0100] The inventors have found that said association can be easily obtained from a natural product by low temperature hydrogenation (35°C maximum) of natural CNSL as reported in the present application in example 3. In particular, there is no need to isolate the hydrogenated anacardic acid contained in the hydrogenated natural CNSL, which would constitute an additional purification step, complicating the preparation of the product.

[0101] In addition, the product obtained is notably in solid form, which facilitates use and storage.

[0102] The amounts of hydrogenated cardol (from 10.0 to 25.0%), hydrogenated methyl cardol (from 0.0 to 5.0%) and hydrogenated cardanol (from 2.0 to 15%) in the claimed concentrations in association with hydrogenated anacardic acid (from 50.0 to 85.0%) do not exhibit any destructive and / or inhibitory effect on the properties related to hydrogenated anacardic acid as demonstrated in the examples.

[0103] Cashew nuts comprise 55-65% shell by weight and 35-45% kernel by weight. Cashew nut shells are therefore an agricultural waste generated by cashew nut processing plants and thus an inexpensive starting material. Cashew nut shells in particular contain a dark reddish-brown caustic oil, in the range of 15-40% by weight, rich in phenolic lipids and have potential for the production of high-value fuels and chemicals, allowing an eco-responsible and local alternative to products from the petroleum industry.

[0104] According to a particular embodiment, the invention relates to the association as defined above, in which

[0105] - hydrogenated anacardic acid represents from 0.0 to 30.0% by total weight of the association,

[0106] - hydrogenated cardol represents from 10.0 to 25.0% by total weight of the association,

[0107] - hydrogenated methyl cardol represents from 0.0 to 5.0% by total weight of the association, and

[0108] - hydrogenated cardanol represents from 50.0 to 85.0% by total weight of the association.

[0109] According to a particular embodiment, the invention relates to the association as defined above, in which said association is derived from a hydrogenated cashew nut shell extract, in particular hydrogenated CNSL, preferably hydrogenated technical CNSL. Advantageously, said association as defined above is derived from a technical CNSL having undergone hydrogenation of the C15 alkyl chains of its phenolic constituents.

[0110] The range “0.0 to 30.0%” includes the following ranges: 0.0 to 1.0%, 1.0 to 2.0%; 2.0 to 3.0%; 3.0 to 4.0%; 4.0 to 5.0%; 5.0 to 6.0%; 6.0 to 7.0%; 7.0 to 8.0%; 8.0 to 9.0%; 9.0 to 10.0%; 10.0 to 11.0%; 11.0 to 12.0%; 12.0 to 13.0%; 13.0 to 14.0%; 14.0 to 15.0%; 15.0 to 16.0%; from 16.0 to 17.0%; from 17.0 to 18.0%; from 18.0 to 19.0%; from 19.0 to 20.0%; from 20.0 to 21.0%; from 21.0 to 22.0%; from 22.0 to 23.0%; from 23.0 to 24.0%; from 24.0 to 25.0%, from 25.0 to 26.0%, from 26.0 to 27.0%, from 27.0 to 28.0%, from 28.0 to 29.0%, from 29.0 to 30.0%.

[0111] Another subject of the present invention relates to the use of a combination as defined above or of hydrogenated anacardic acid as a food supplement for reducing methanogenesis and / or for improving digestive fermentation, in particular as a food supplement for ruminants.

[0112] According to a particular embodiment, the invention relates to the use of a combination as defined above or of hydrogenated anacardic acid as a food supplement for reducing methanogenesis and for improving digestive fermentation, in particular as a food supplement for ruminants.

[0113] According to a particular embodiment, the invention relates to the use of a combination as defined above or of hydrogenated anacardic acid as a food supplement for reducing methanogenesis, in particular as a food supplement for ruminants.

[0114] According to a particular embodiment, the invention relates to the use of a combination as defined above or of hydrogenated anacardic acid as a food supplement for improving digestive fermentation, in particular as a food supplement for ruminants.

[0115] According to a particular embodiment, the invention relates to the use of a combination as defined above as a food supplement for reducing methanogenesis, in particular as a food supplement for ruminants.

[0116] According to a particular embodiment, the invention relates to the use of hydrogenated anacardic acid as a food supplement for reducing methanogenesis, in particular as a food supplement for ruminants. Another subject of the present invention relates to the use of a combination as defined above or of hydrogenated anacardic acid for the manufacture of a food supplement for reducing methanogenesis and / or for improving digestive fermentation.

[0117] According to a particular embodiment, the invention relates to the use as defined above or of hydrogenated anacardic acid for the manufacture of a food supplement for reducing methanogenesis.

[0118] According to a particular embodiment, the invention relates to the use as defined above or of hydrogenated anacardic acid for the manufacture of a food supplement for improving digestive fermentation.

[0119] According to a particular embodiment, the invention relates to the use as defined above or of hydrogenated anacardic acid for the manufacture of a food supplement for reducing methanogenesis and for improving digestive fermentation.

[0120] According to a particular embodiment, the invention relates to the use as defined above of an association as defined above for the manufacture of a food supplement for reducing methanogenesis.

[0121] According to a particular embodiment, the invention relates to the use as defined above of hydrogenated anacardic acid for the manufacture of a food supplement for reducing methanogenesis.

[0122] Another subject of the present invention relates to the use as defined above of an association as defined above or of hydrogenated anacardic acid, said association or hydrogenated anacardic acid being used at a rate of 50 to 600 ppm, preferably 50 to 500 ppm.

[0123] According to a particular embodiment, the invention relates to the use as defined above of an association as defined above, said association being used at a rate of 50 to 600 ppm, preferably 50 to 500 ppm.

[0124] According to a particular embodiment, the invention relates to the use as defined above of hydrogenated anacardic acid, the latter being used at a rate of 50 to 600 ppm, preferably 50 to 500 ppm.

[0125] Another subject of the present invention relates to a food comprising the combination as defined above or hydrogenated anacardic acid, at a rate of 50 to 600 ppm, preferably 50 to 500 ppm. According to a particular embodiment, the invention relates to a food as defined above comprising the combination as defined above, at a rate of 50 to 600 ppm, preferably 50 to 500 ppm.

[0126] According to a particular embodiment, the invention relates to a food as defined above comprising hydrogenated anacardic acid, at a rate of 50 to 600 ppm, preferably 50 to 500 ppm.

[0127] Another subject matter of the present invention relates to a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV): or a composition comprising or consisting of hydrogenated anacardic acid, for use against digestive pathological conditions selected from abdominal tympanism or coccidiosis. According to a particular embodiment, the invention relates to a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV): in which

[0128] - hydrogenated anacardic acid represents from 50.0 to 85.0% by total weight of the association,

[0129] - hydrogenated cardol represents from 10.0 to 25.0% by total weight of the association, - hydrogenated methyl cardol represents from 0.0 to 5.0% by total weight of the association, and

[0130] - hydrogenated cardanol represents from 2.0 to 15.0% by total weight of the association, or composition comprising or consisting of hydrogenated anacardic acid, for its use against digestive pathological conditions chosen from abdominal tympanism or coccidiosis. According to a particular embodiment, the invention relates to a composition for its use as defined above for non-human animals.

[0131] According to a particular embodiment, the invention relates to a composition for its use as defined above against abdominal tympanism.

[0132] According to a particular embodiment, the invention relates to a composition for its use as defined above against coccidiosis.

[0133] According to a particular embodiment, the invention relates to a composition for its use as defined above, in which:

[0134] - hydrogenated anacardic acid represents from 50.0 to 85.0% by total weight of the association,

[0135] - hydrogenated cardol represents from 10.0 to 25.0% by total weight of the association,

[0136] - hydrogenated methyl cardol represents from 0.0 to 5.0% by total weight of the association, and

[0137] - hydrogenated cardanol represents from 2.0 to 15.0% by total weight of the association.

[0138] According to a particular embodiment, the invention relates to a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract for use against coccidiosis.

[0139] According to a particular embodiment, the invention relates to a composition of natural or technical hydrogenated cashew nut shell liquid (hydrogenated CNSL) for use against coccidiosis.

[0140] According to a particular embodiment, the invention relates to a composition of hydrogenated natural cashew nut shell liquid (hydrogenated natural CNSL) for use against coccidiosis.

[0141] According to a particular embodiment, the invention relates to a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV):

[0142] in which

[0143] - hydrogenated anacardic acid represents from 50.0 to 85.0% by total weight of the association,

[0144] - hydrogenated cardol represents from 10.0 to 25.0% by total weight of the association, - hydrogenated methyl cardol represents from 0.0 to 5.0% by total weight of the association, and

[0145] - hydrogenated cardanol represents from 2.0 to 15.0% by total weight of the association, or composition comprising or consisting of hydrogenated anacardic acid, for its use against coccidiosis.

[0146] The inventors have surprisingly found that hydrogenated natural CNSL has an effect on the viability of sporozoites, namely the pathogen of coccidiosis. In particular, hydrogenated natural CNSL is more effective than natural CNSL and technical CNSL against the viability of sporozoites. Natural CNSL is more effective than technical CNSL. According to a particular embodiment, the invention relates to a composition of hydrogenated technical cashew nut shell liquid (hydrogenated technical CNSL) for use against coccidiosis.

[0147] According to a particular embodiment, the invention relates to a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV): in which

[0148] - hydrogenated anacardic acid represents from 0.0 to 30.0% by total weight of the association,

[0149] - hydrogenated cardol represents from 10.0 to 25.0% by total weight of the association,

[0150] - hydrogenated methyl cardol represents from 0.0 to 5.0% by total weight of the association, and

[0151] - hydrogenated cardanol represents from 50.0 to 85.0% by total weight of the combination, for its use against coccidiosis. The inventors have also unexpectedly and surprisingly found that the hydrogenated technical CNSL also has an effect on the viability of sporozoites, namely the pathogen of coccidiosis. In particular, the hydrogenated technical CNSL is more effective than the natural CNSL and than the technical CNSL against the viability of sporozoites and the invasion of cells by E. tennella sporozoites.

[0152] According to a particular embodiment, the invention relates to a composition for its use as defined below against coccidiosis, said composition being at a dose of 5.00 to 1000 pg by weight of said composition in 1 mL of an acceptable pharmaceutical medium.

[0153] The range of "5.00 to 1000 pg" includes the following ranges: 5.00 to 6.25 pg; 6.25 to 12.5 pg; 12.5 to 25 pg; 25 to 50 pg; 50 to 100 pg; 100 to 200 pg; 200 to 300 pg; 300 to 400 pg; 400 to 500 pg; 500 to 600 pg; 600 to 700 pg; 800 to 800 pg; 800 to 900 pg; 900 to 1000 pg.

[0154] Another object of the present invention relates to a non-therapeutic method for decreasing methanogenesis in ruminants, comprising administering a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV):

[0155] or a composition comprising or consisting of hydrogenated anacardic acid.

[0156] According to a particular embodiment, the invention relates to a non-therapeutic method as defined above, for reducing methanogenesis in ruminants, comprising the administration of a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV):

[0157]

[0158] According to a particular embodiment, the invention relates to a non-therapeutic method as defined above, for reducing methanogenesis in ruminants, comprising the administration of a composition comprising or consisting of hydrogenated anacardic acid.

[0159] FIGURES AND EXAMPLES

[0160] Figure 1 represents the total production of volatile fatty acids (VFAs) produced after 72 hours of incubation in an in vitro digestive simulation medium containing a dose of 200, 400 and 600 pg / mL for natural CNSL, technical CNSL, hydrogenated natural CNSL and a control, respectively.

[0161] Figure 2 represents the production of propionate as a percentage of VFA after 72 hours of incubation in an in vitro digestive simulation medium containing a dose of 200, 400 and 600 pg / mL for natural CNSL, technical CNSL, hydrogenated natural CNSL and a control, respectively. Figure 3 represents the production of valerate as a percentage of VFA after 72 hours of incubation in an in vitro digestive simulation medium containing a dose of 200, 400 and 600 pg / mL for natural CNSL, technical CNSL, hydrogenated natural CNSL and a control, respectively.

[0162] Figure 4 represents the acetate production as a percentage of VFA after 72 hours of incubation in an in vitro digestive simulation medium containing a dose of 200, 400 and 600 pg / mL for natural CNSL, technical CNSL, hydrogenated natural CNSL and a control, respectively.

[0163] Figure 5 represents the butyrate production as a percentage of VFA after 72 hours of incubation in an in vitro digestive simulation medium containing a dose of 200, 400 and 600 pg / mL for natural CNSL, technical CNSL, hydrogenated natural CNSL and a control, respectively.

[0164] Figure 6 represents the digestibility of organic matter in percentage after 72 hours of incubation in an in vitro digestive simulation medium containing respectively a dose of 200, 400 and 600 pg / mL for natural CNSL, technical CNSL, hydrogenated natural CNSL and a control.

[0165] Figure 7 represents the pH of the medium after 72 hours of incubation in an in vitro digestive simulation medium containing respectively a dose of 200, 400 and 600 pg / mL for natural CNSL, technical CNSL, hydrogenated natural CNSL and a control.

[0166] Figure 8 represents the methane production in mL per gram of organic matter (mL / g MO) as a function of incubation time in an in vitro digestive simulation medium containing respectively a dose of 200, 400 and 600 pg / mL for natural CNSL (a), technical CNSL (b), hydrogenated natural CNSL (c) and a control.

[0167] Figure 9 represents the total gas production in mL per gram of organic matter (mL / g OM) as a function of incubation time in an in vitro digestive simulation medium containing respectively a dose of 200, 400 and 600 pg / mL for natural CNSL (a), technical CNSL (b), hydrogenated natural CNSL (c) and a control.

[0168] Figure 10 represents the percentage of methane produced relative to total gas production as a function of incubation time in an in vitro digestive simulation medium containing respectively a dose of 200, 400 and 600 pg / mL for natural CNSL (a), technical CNSL (b), hydrogenated natural CNSL (c) and a control.

[0169] Figure 11 represents the viability data of Eimeria sporozoites exposed to 8 doses (from 6.25 to 800 pg / mL) of product A, product A corresponding to a hydrogenated natural CNSL resulting from solvent extraction; the upper part represents the histogram of the data compared to positive controls (amprolium and monensin at 50 pg / mL), the bars represent the means ± standard error of the mean. *p<0.05, **p<0.01, ***p<0.001 vs. live control without treatment; the lower part represents the viability of sporozoites as a function of the logarithm of the doses (from 6.25 to 800 pg / mL) in product A, as well as the associated nonlinear regression which allows to determine the half-maximal inhibitory concentration (ECso), the bars represent the means ± standard error of the mean.

[0170] Figure 12 represents the viability data of Eimeria sporozoites exposed to 8 doses (from 6.25 to 800 pg / mL) of product B, product B corresponding to a hydrogenated natural CNSL from press extraction; the upper part represents the histogram of the data compared to positive controls (amprolium and monensin at 50 pg / mL), the bars represent the means ± standard error of the mean. *p<0.05, **p<0.01, ***p<0.001 vs. live control without treatment; the lower part represents the viability of sporozoites as a function of the logarithm of the doses (from 6.25 to 800 pg / mL) in product B, as well as the associated nonlinear regression which allows to determine the half-maximal inhibitory concentration (EC50), the bars represent the means ± standard error of the mean.

[0171] Figure 13 represents the viability data of Eimeria sporozoites exposed to 8 doses (from 6.25 to 800 pg / mL) of product C, product C corresponding to a natural CNSL from press extraction; the upper part represents the histogram of the data compared to positive controls (amprolium and monensin at 50 pg / mL), the bars represent the means ± standard error of the mean. *p<0.05, **p<0.01, ***p<0.001 vs. live control without treatment; the lower part represents the viability of sporozoites as a function of the logarithm of the doses (from 6.25 to 800 pg / mL) in product C, as well as the associated nonlinear regression which allows to determine the half-maximal inhibitory concentration (EC50), the bars represent the means ± standard error of the mean.

[0172] Figure 14 represents the viability data of Eimeria sporozoites exposed to 8 doses (from 6.25 to 800 pg / mL) of product D, product D corresponding to a natural CNSL resulting from solvent extraction; the upper part represents the histogram of the data compared to positive controls (amprolium and monensin at 50 pg / mL), the bars represent the means ± standard error of the mean. *p<0.05, **p<0.01, ***p<0.001 vs. live control without treatment; the lower part represents the viability of sporozoites as a function of the logarithm of the doses (from 6.25 to 800 pg / mL) in product D, as well as the associated nonlinear regression which allows to determine the half-maximal inhibitory concentration (EC50), the bars represent the means ± standard error of the mean.Figure 15 represents the viability data of Eimeria sporozoites exposed to 8 doses (from 6.25 to 800 pg / mL) of product E, product E corresponding to a technical CNSL; the upper part represents the histogram of the data compared to positive controls (amprolium and monensin at 50 pg / mL), the bars represent the means ± standard error of the mean. *p<0.05, **p<0.01, ***p<0.001 vs. live control without treatment; the lower part represents the viability of sporozoites as a function of the logarithm of the doses (from 6.25 to 800 pg / mL) in product E, as well as the associated nonlinear regression which allows to determine the half-maximal inhibitory concentration (ECso), the bars represent the means ± standard error of the mean.

[0173] Figure 16 represents the viability data of Eimeria sporozoites exposed to 8 doses (from 6.25 to 800 pg / mL) of product F, product F corresponding to a hydrogenated technical CNSL; the upper part represents the histogram of the data compared to positive controls (amprolium and monensin at 50 pg / mL), the bars represent the means ± standard error of the mean. *p<0.05, **p<0.01, ***p<0.001 vs. live control without treatment; the lower part represents the viability of sporozoites as a function of the logarithm of the doses (from 6.25 to 800 pg / mL) in product F, as well as the associated nonlinear regression which allows to determine the half-maximal inhibitory concentration (ECso), the bars represent the means ± standard error of the mean.

[0174] Figure 17 shows the data (in percentage), in the form of histograms A, B, C, D, E and F, of the invasion capacity of E. tenella sporozoites exposed to 8 doses (from 6.25 to 800 pg / mL) of products A, B, C, D, E and F and compared to positive controls (amprolium and monensin at 50 pg / mL); product A corresponding to a natural hydrogenated CNSL from solvent extraction, product B corresponding to a natural hydrogenated CNSL from press extraction, product C corresponding to a natural CNSL from press extraction, product D corresponding to a natural CNSL from solvent extraction, product E corresponding to a technical CNSL and product F corresponding to a technical hydrogenated CNSL, the bars represent the means ± standard error of the mean. *p<0.05,**p<0.01, ***p<0.001 vs. living control without treatment.

[0175] Figure 18 represents, in the form of graphs A, B, C, D, E and F, the invasion capacity of E. tenella sporozoites as a function of the logarithm of the doses (from 6.25 to 800 pg / mL) in products A, B, C, D, E and F respectively; product A corresponding to a hydrogenated natural CNSL from solvent extraction, product B corresponding to a hydrogenated natural CNSL from press extraction, product C corresponding to a natural CNSL from press extraction, product D corresponding to a natural CNSL from solvent extraction, product E corresponding to a technical CNSL and product F corresponding to a hydrogenated technical CNSL, as well as the associated non-linear regression which allows the determination of the half-maximal inhibitory concentration (ECso), the bars represent the means ± the standard error of the mean.

[0176] Example 1: Extraction of natural CNSL from cashew nut shells

[0177] Natural CNSL was obtained from cashew nut shells by a solvent extraction process. The required mass of previously ground cashew nut shells was suspended in the desired solvent (ethyl acetate, ratio: m CO ques / V so ivant = 1 / 2.5) at 50°C for 3 h. The suspension was then filtered, the shells were washed with solvent, and then the filtrates were combined. Natural CNSL was obtained after removal of the solvent.

[0178] Thus, 160 kg of previously crushed cashew shells were introduced into a filter bottom tank equipped with a felt filter cloth (apparent porosity of approximately 25 μm). 360 kg of ethyl acetate (400 L) were then added to the tank. The suspension was then stirred at 50°C for 3 h. Vacuum filtration was then carried out to separate the filtrate from the extracted shell residues. 216 kg of ethyl acetate (240 L) were then introduced into the filter bottom tank and the suspension was again stirred at 50°C for 30 minutes. A second vacuum filtration was carried out to separate the filtrate from the extracted shell residues. The two filtrates were then combined, and the solvent was removed under reduced pressure using a falling film concentrator, at an evaporation temperature between 25 and 50°C.56.6 kg of natural CNSL were thus obtained (black oil, the mass percentage of ethyl acetate was 13.4%, yield r = 31% excluding ethyl acetate).

[0179] RM N- 1 H (400 M Hz, CDCh): 7.35 (t, J = 7.9 Hz, H aci of anacardium), 7, 13 (t, J = 7.6 Hz, H cardano i) , 6.88 - 6.86 (m, H aC anacardial ide), 6.78 - 6.74 (m, H ac anacardic ide and Hcardanol), 6.66 - 6.63 (m, Hcardanol), 6.24 (m, Hcardol and Hmethyl cardol), 6, 18 (S, Hcardol), 5.87 5.76 (m, Hc=c), 5.46 5.30 (m, Hc=c), 5.07 — 4.96 (m, Hc=c), 2.99 — 2.96 (m, H a anacardic cide), 2.82 — 2.76 (m, Hside chain), 2.57 2.53 (m, Hcardanol), 2.50 2.43 (m, Hcardol and Hmethyl cardol), 2.10 (S, Hmethyl cardol), 2.11 2.00 (m,

[0180] Hside chain), 1.63 1.53 (m, Hside chain), 1.40 1.25 (m, Hside chain), 0.93 0.86 (m, Hside chain).

[0181] This CNSL was dried under high vacuum to remove ethyl acetate (% r ési uei < 2% by mass). The mass percentages of the phenolic compounds of interest in natural CNSL are reported in the following table 1.

[0182] Table 1: Mass percentages of phenolic compounds in natural CNSL after the extraction solvent removal step.

[0183] Example 2: Decarboxylation of natural CNSL into technical CNSL

[0184] 80.2 g of natural CNSL were dissolved in 60 mL of refluxing o-xylene (temperature between 140 and 150°C). The medium was left at reflux with stirring for 5 h 10 min (kinetic monitoring of the decarboxylation by TLC, eluent cyclohexane / ethyl acetate 50 / 50, developer KMnCL). The medium was filtered through filter paper, then the solvent was removed under reduced pressure (rotary evaporator) at 60°C. 74.8 g of technical CNSL were thus obtained in the form of a black viscous oil. The process was repeated until the desired quantity of technical CNSL was reached (2983 g).

[0185] NMR- 1H (400 MHz, CDCI3): 7.13 - 7.10 (m, cardanol), 6.77 - 6.74 (m, cardanol), 6.67 - 6.63 (m, cardanol), 6.24 (m, cardol and methyl cardol), 6.17 (t, cardol), 5.87 - 5.77 (m, alkene), 5.43 - 5.33 (m, alkene), 5.08 - 4.97 (m, alkene), 2.84 - 2.76 (m, side chain), 2.57 - 2.53 (m, cardanol), 2.50 - 2.46 (m, cardol and methyl cardol), 2.07 - 1.90 (m, side chain), 1.62 - 1.55 (m, side chain), 1 ,35 - 1 ,27 (m, side chain), 0.93 - 0.87 (m, side chain).

[0186] This technical CNSL was dried under high vacuum to remove xylene (% r residue < 1% by mass). The mass percentages of the phenolic compounds of interest in the technical CNSL are reported in the following table 2.

[0187] Table 2: Mass percentages of phenolic compounds in technical CNSL after the decarboxylation solvent removal step.

[0188] Example 3: Preparation of hydrogenated natural CNSL

[0189] Materials and methods

[0190] The operating setup consists of a 1 L flask, a magnetic bar, a heating stirring plate and an oil bath.

[0191] Natural CNSL was produced internally by ORPIA according to Example 1.

[0192] The Pd / C (5%) comes from Fisher Scientific and the 10% from Sigma Aldrich.

[0193] Ethanol (96%) is supplied by VWR. Hydrogenation operating protocol

[0194] Product 1

[0195] Approximately 200 g of CNSL were solubilized in 400 mL of ethanol, then 8 g of Pd / C (4% by mass relative to CNSL) were added to the medium. The latter was stirred at room temperature, the medium was first purged under nitrogen, then placed under dihydrogen. The suspension was left stirring and under dihydrogen atmosphere at a temperature of 20 to 35°C for 1 day to a week. The kinetic monitoring of the hydrogenation was carried out by NMR 1 H.

[0196] The suspension was filtered using a Whatman® glass microfiber filter. The solid residue was rinsed with ethyl acetate, and the solvent mixture was removed under reduced pressure (rotary evaporator).

[0197] The residue was dried at 50 °C with stirring (using the rotary evaporator) at 25 rpm, protected from light, under a vane pump for 15 h.

[0198] A mass of 167.7 g of hydrogenated CNSL was obtained.

[0199] Product 2

[0200] 53.19 g of CNSL were solubilized in 250 mL of ethanol, then 1.97 g of Pd / C (3.7% by mass relative to CNSL) were added to the medium. The latter was stirred at room temperature after being purged under nitrogen, then placed under dihydrogen. The suspension was left stirring and under dihydrogen atmosphere at room temperature overnight.

[0201] The suspension was filtered using a Whatman® glass microfiber filter, and the solvent was removed under reduced pressure (rotary evaporator).

[0202] The residue was left to dry under a vane pump for 15 h.

[0203] A mass of 93.5 g of hydrogenated CNSL was obtained as a beige solid.

[0204] Characterizations of hydrogenated natural CNSL

[0205] NMR spectrum of Product 1

[0206] NMR- 1 H (400 MHz, acetone-d6): 7.33 (1 H, t, AA), 7.07 (1 H, t, cardanol), 6.78 (2H, d, AA), 6.68

[0207] - 6.62 (3H, m, cardanol), 6.23 (2H, s, MC), 6.19 - 6.16 (3H, m, cardol), 3.00 - 2.96 (2H, m, AA), 2.54 - 2.51 (2H, m, cardanol), 2.46 - 2.38 (2H, m, cardol and MC), 2.03 (3H, s, MC), 1.65

[0208] - 1.54 (m, side chain), 1.34 - 1.27 (m, side chain), 0.89 - 0.86 (m, side chain). Traces of residual solvent (< 1% by mass).

[0209] The mass percentages of the phenolic compounds of interest in the hydrogenated natural CNSL of product 1 are indicated in the following table 3:

[0210] Table 3: Mass percentages of phenolic compounds in hydrogenated natural CNSL of product 1.

[0211] NMR Spectrum of Product 2 NMR- 1 H (400 MHz, acetone-d6): 7.33 (1 H, t, AA), 7.07 (1 H, t, cardanol), 6.78 (1 H, d, AA), 6.68

[0212] - 6.62 (3H, m, cardanol), 6.23 (2H, s, MC), 6.19 - 6.16 (3H, m, cardol), 3.00 - 2.96 (2H, m, AA), 2.54 - 2.51 (2H, m, cardanol), 2.46 - 2.38 (2H, m, cardol and MC), 2.03 (3H, s, MC), 1.65

[0213] - 1.54 (m, side chain), 1.34 - 1.27 (m, side chain), 0.89 - 0.86 (m, side chain). Traces of residual solvent (< 1%) The mass percentages of the phenolic compounds of interest in the hydrogenated natural CNSL of product 2 are indicated in the following table 4:

[0214] Table 4: Mass percentages of phenolic compounds in hydrogenated natural CNSL of product 2.

[0215] Example 4: In vitro study of the methanogenesis reduction potential of hydrogenated natural CNSL compared to natural and technical CNSL

[0216] The following in vitro experiments were conducted to investigate the methanogenesis-mitigating potential, i.e., the reduction of methane (CH4) formation, of hydrogenated cashew nut shell liquid or extract (hydrogenated CNSL) compared to natural and technical grade CNSL. They also aimed to analyze the potential dose-dependent response of the three different forms of CNSL. The experiments were implemented by simulating digestive conditions using an artificial rumen derived from the ruminal fluid of dairy cows. Three types of CNSL (i.e., natural, technical grade, and hydrogenated natural) were dissolved in ethanol at the target dose with the substrate and incubated for 72 hours in ruminal fluid from three Holstein Friesian dairy cows that was pooled, filtered, and buffered.The assays tested were 200, 400, and 600 pg / mL for each of the three CNSLs (natural, technical, and hydrogenated natural). A control treatment, containing only the substrates and no CNSL, was also included. After inoculation, the different fermentation media, each enclosed in a bottle, were immediately connected to an analysis device (APES) to measure the total cumulative gas production (GP). During incubation, 12 gas samples were collected from each bottle at 0, 2, 4, 6, 8, 12, 24, 30, 36, 48, 60, and 72 h of incubation and analyzed for methane (CH4) content.

[0217] Example 5 - Materials and methods of the in vitro study

[0218] 5. 1. Experimental design

[0219] The in vitro study to determine the methanogenesis mitigation potential of the three different types of natural, technical and hydrogenated natural CNSL was carried out in an animal nutrition research laboratory.

[0220] Ruminal fluid from fistulated dairy cows served as inoculum for the in vitro study.

[0221] A total of nine combinations were tested corresponding to the association of a CNSL and a dose, as illustrated in Table 5.

[0222] Table 5: Combinations tested corresponding to the association of a CNSL and a dose.

[0223] 5.2. Ruminal fluid

[0224] Three dairy cows were fitted with a permanent ruminal cannula (10 cm inner diameter, type 1C) to serve as ruminal fluid donors for the in vitro experiment. These cows received a total mixed ration consisting of grass silage, corn silage, and concentrate. The handling of the dairy cows was approved by an ethics committee and in accordance with the legislation of the study country on the use of laboratory animals. Rumen fluid was collected in three equal volumes from the front, mid-ventral sac, and caudodorsal region of the rumen using the method described by van Zijderveld et al. (J. Dairy Sci. 94, 1445-1454. 2011). After collecting the rumen fluid from each dairy cow, it was transferred into preheated thermos flasks (39°C), previously filled with CO2.Ruminal fluid collected from each dairy cow was pooled and 600 pL samples were taken to determine volatile fatty acid (VFA) concentrations. The rumen fluid was then filtered through 2 layers of cloth to remove large rumen fluid particles and mixed with pre-warmed anaerobic buffer (39. 0 C) / a mineral solution (1:2, v / v) according to Cone et al. (Anim. Feed Sci. Technol. 172:34-41 1996).

[0225] 5.3. In vitro study

[0226] Gas production (GP) was determined using fully automated GP equipment as described in Cone et al. (Anim. Feed Sci. Technol. 172:34-41 1996). The substrates, grass silage and maize silage, were ground to pass a 1 mm sieve using a cross-beater mill (Peppink 100 AN, Olst, The Netherlands). The chemical composition of the substrates is shown in Table 6.

[0227] Approximately 0.5 g of dry matter (DM) substrate (i.e., 0.25 g of DM grass silage and 0.25 g of DM corn silage) was used as substrate for each fermentation bottle. CNSL types (i.e., natural, technical grade, and hydrogenated natural CNSL) were dissolved at the desired dose (targeting 60 mL) in ethanol with the substrate and dried to evaporate the ethanol within 48 hours at room temperature. The combined CNSL and substrate were then incubated in 250 mL fermentation bottles (Schott, Mainz, Germany). Each experimental treatment (i.e., CNSL type x dose) as well as the control (i.e., ruminal fluid with substrate only) were included in triplicate in fermentation bottles, with blanks (i.e., ruminal fluid without substrate) included in duplicate.

[0228] The fermentation bottles were pre-rinsed with CO2 and placed in a shaking water bath, maintained at 39 °C with 40 movements per minute. Then, the bottles were inoculated with 60 mL of filtered and buffered ruminal fluid and connected to the fully automated equipment (Cone et al., 1996). Before gas measurements began, the fermentation bottles were fitted with a glass extension and sealed with a screw cap fitted with an airtight septum. The screw caps had a small opening to allow the passage of a fine needle. At distinct incubation times (i.e., 0, 2, 4, 6, 8, 12, 24, 30, 36, 48, 60, and 72 hours of incubation), 10 pL aliquots of the headspace gas were collected through this opening using a gas-tight syringe (Hamilton 1701 N, Point five needle style, 51 mm; Hamilton, Bonaduz, Switzerland).Immediately afterward, the collected headspace gas samples were directly injected into the injection port of the gas chromatography (GC; GC8000Top CE instruments, Milan, Italy) to measure the methane (CH4) concentration in the headspace gas samples, as described by Pellikaan et al. (Anim. Feed Sci. Technol. 168:196-205, 2011) and to quantify the cumulative methane production as described by Hatew et al. (Grass Forage Sci. 70:474-490, 2014). After 72 h of incubation, the fermentation was stopped and 600 pL fermentation samples were collected from each bottle to determine the VFA concentrations of the fermentation liquid as described by Van Gastelen et al. (J. Dairy Sci. 104:4174-4191, 2021). The organic matter (OM) contained in the substrate residues of the fermentation bottles was analyzed to determine the level of digestibility of the OM.

[0229] Table 6: Chemical composition (in g / kg of dry matter) of grass silage and corn silage used as substrates. 5.4. Chemical analyses

[0230] Grass silage and corn silage substrates were analyzed for dry matter (DM), ash, nitrogen (N), starch, crude fat, neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL). Substrate residues in fermentation bottles after 72 h of incubation were analyzed for ash. The wet chemistry methods performed are described by Abrahamse et al. (J. Dairy Sci. 91:2033-2045, 2008). Calorimetry measured by bomb calorimeter (ISO 9831; International Organization for Standardization, 1998) was used to determine gross energy (GE) content. Crude protein was calculated as N x 6.25, with N determined using the Dumas method (ISO 16634-1; International Organization for Standardization, 2008). Organic matter was calculated as: (1000 - ash).Ruminal fluid samples (i.e., before incubation and after 72 hours of incubation) were analyzed for VFAs as described by van Gastelen et al. (J. Dairy Sci. 104:4174-4191, 2021).

[0231] Example 6 - Results - Volatile Fatty Acids (VFA)

[0232] The VFAs after 72 hours of in vitro incubation with different doses of the different types of CNSL are presented: in Table 7 for natural CNSL, in Table 8 for technical CNSL, in Table 9 for hydrogenated natural CNSL and in Figures 1 to 5.

[0233] In the presented results, the branched-chain volatile fatty acid (VFA) content is defined as the sum of iso-valerate and iso-butyrate compounds.

[0234] The "NGR" index corresponding to the ratio between non-glycogenic AGV and glycogenic AGV is defined as follows (Cone and Becker et al., Anim. Feed Sci. Technol. 172:34-41): acetate + 2 x butyrate + 2 x isobutyrate + valerate + isovalerate propionate + valerate + isovalerate

[0235] Table 7: Effect of natural CNSL at different doses on volatile fatty acids (VFAs) after 72 hours of in vitro incubation.

[0236] Table 8: Effect of technical CNSL at different doses on volatile fatty acids (VFAs) after 72 hours of in vitro incubation.

[0237] Table 9: Effect of hydrogenated natural CNSL at different doses on volatile fatty acids (VFAs) after 72 hours of in vitro incubation.

[0238] Example 7 - Results - pH and digestibility of organic matter (OM) The pH and digestibility of organic matter (OM) after 72 hours of in vitro incubation with different doses of the different types of CNSL are presented in Table 10 and Figures 6 and 7.

[0239] Table 10: pH and digestibility of organic matter (OM) after 72 hours of in vitro incubation with different doses of different types of CNSL.

[0240] Example 8 - Results - Production of gas and methane

[0241] The total volume (mL) of gas (GP) and methane (ChL) generated during a 72-hour in vitro incubation per gram of initial weight of MO substrate as well as the methane concentration in the total gases produced are shown in Table 11. These represent only the cumulative values ​​at the end of 72 hours of incubation.

[0242] The profiles of the same variables, i.e. gas production (GP) and methane production (ChL), in mL / g of organic matter (OM), as well as the concentration of methane (CH4) in total gases, in percentage (%), throughout the 72 hours of incubation, are presented respectively in Figures 8 to 10.

[0243] Table 11: Total volume (mL) of gas (GP) and methane (CH4) generated during a 72-hour in vitro incubation (per gram of initial weight of MO substrate) as well as the concentration of methane in the total gases produced (in %).

[0244] Example 9 - Analysis of results

[0245] Methanogenesis

[0246] Based on the results obtained, hydrogenated natural CNSL can effectively reduce methanogenesis. When 200, 400 or 600 pg / mL of hydrogenated natural CNSL were incubated, CH4 methane production (mL / g of OM) decreased by 37%, 51% and 63%, respectively, compared to the control.

[0247] The methanogenesis mitigation levels are between those of natural CNSL and technical-grade CNSL. Hydrogenated natural CNSL is thus slightly less effective than natural CNSL, but more effective than technical-grade CNSL.

[0248] Similar to natural CNSL and technical CNSL, a dose-response effect in which the inhibition of methane production increased with increasing dose of hydrogenated natural CNSL was observed.

[0249] AGV, OM digestibility, pH and gas production

[0250] Similar to natural CNSL, hydrogenated natural CNSL leads to an increase in the level of propionate and valerate at the expense of acetate and butyrate. A minimal effect of hydrogenated natural CNSL on the total volatile fatty acid (VFA) content is observed. Indeed, a difference of at most - 4% is observed compared to the control for hydrogenated natural CNSL, while for natural CNSL the difference reaches more than 20% compared to the control.

[0251] A minimal effect of hydrogenated natural CNSL on the pH and consequently on the acidity of the digestive environment is observed, whereas natural CNSL causes a variation in the acidity of the digestive environment.

[0252] The effect of hydrogenated natural CNSL on organic matter (OM) digestibility is lower than that of natural CNSL and technical CNSL. In this study, a small effect (difference of the order of -5% with the control) on organic matter digestibility was observed for hydrogenated natural CNSL while natural CNSL resulted in a digestibility difference of up to -28.8% compared to the control.

[0253] The effect of hydrogenated natural CNSL on gas production (GP) is lower than that of natural CNSL, but higher than that of technical CNSL.

[0254] In conclusion, hydrogenated natural CNSL allows a reduction in gas production, particularly in methanogenesis, a shift towards the production of propionate and valerate to the detriment of acetate and butyrate, with minimal and attenuated side effects on fermentation, by promoting the digestibility of organic matter and the maintenance of pH.

[0255] Example 10 - Preparation of CNSLs tested for in vitro evaluation of anticoccidial effect

[0256] 10.1 - Natural solvent-extracted CNSL (sample D)

[0257] Natural CNSL was obtained from cashew nut shells by a solvent extraction process according to Example 1. The required mass of previously ground cashew nut shells was suspended in the desired solvent (ethyl acetate, ratio of: m CO ques / Vsoivant = 1 / 2.5) at 50°C for 3 h. The suspension was then filtered, the shells were washed with solvent, and then the filtrates were combined. Natural CNSL was obtained after removal of the solvent.

[0258] Thus, 160 kg of previously crushed cashew shells were introduced into a filter bottom tank equipped with a felt filter cloth (apparent porosity of approximately 25 μm). 360 kg of ethyl acetate (400 L) were then added to the tank. The suspension was then stirred at 50°C for 3 h. Vacuum filtration was then carried out to separate the filtrate from the extracted shell residues. 216 kg of ethyl acetate (240 L) were then introduced into the filter bottom tank and the suspension was again stirred at 50°C for 30 minutes. A second vacuum filtration was carried out to separate the filtrate from the extracted shell residues. The two filtrates were then combined, and the solvent was removed under reduced pressure using a falling film concentrator, at an evaporation temperature between 25 and 50°C. 56.6 kg of natural CNSL were thus obtained.A fraction of this CNSL was extensively dried under vacuum to remove residual solvent.

[0259] NMR- 1 H (400 MHz, CDCh): 11.03 (wide signal, OH), 7.38 - 7.34 (1 H, m, AA), 7.14 (1 H, t, J = 7.7 Hz, cardanol), 6.88 - 6.86 (1 H, m, AA), 6.79 - 6.74 (2H, m, AA and cardanol), 6.66 - 6.63 (2H, m, cardanol), 6.24 (2H, m, cardol and MC), 6.18 (1 H, t, J = 2.3 Hz, cardanol), 5.85 - 5.78 (m, C=C), 5.47 - 5.30 (m, C=C), 5.08 - 4.96 (m, C=C), 2.99 - 2.96 (m, AA), 2.84 - 2.76 (m, side chain), 2.57 -2.53 (2H, m, cardanol), 2.50 - 2.39 (2H, m, cardol and MC), 2.10 (3H, s, MC), 2.09 - 1.99 (m, side chain), 1.87 (3H, s, MC), 1.40 - 1.25 (m, side chain), 0.93 - 0.86 (m, side chain). Traces of ethyl acetate (< 1% by mass).

[0260] The mass percentages of the phenolic compounds of interest in natural CNSL extracted by solvent are reported in the following table 12:

[0261] Table 12: Mass percentages of phenolic compounds in natural CNSL extracted by solvent (sample D).

[0262] 10.2 - Natural CNSL extracted by solvent then hydrogenated (sample A)

[0263] 53.19 g of the natural CNSL extracted by solvent described previously in paragraph 10.1 were solubilized in 250 mL of ethanol, then 1.99 g of Pd / C (3.7% by mass relative to the CNSL) were added to the medium. The latter was stirred at room temperature, then it was conditioned under nitrogen, and then under dihydrogen. The suspension was left stirring and under dihydrogen atmosphere at room temperature for 22h15 (the kinetic monitoring of the hydrogenation is carried out by NMR 1H). The suspension was filtered through a Millipore filter, and the solvent was then removed using a rotary evaporator. The solid was finally dried under high vacuum at room temperature (20-30°C). 50.31 g of solvent-extracted and then hydrogenated CNSL were obtained, giving a yield of 94.6% (with r =

[0264] NMR- 1 H (400 MHz, acetone-d6): 7.33 (1 H, t, J = 7.9 Hz, AA), 7.07 (1 H, t, J = 7.8 Hz, cardanol), 6.79 - 6.77 (2H, m, AA), 6.67 - 6.62 (3H, m, cardanol), 6.23 (2H, s, MC), 6.19 - 6.16 (3H, m, cardol), 3.00 - 2.96 (2H, m, AA), 2.55 - 2.51 (2H, m, cardanol), 2.46 - 2.37 (2H, m, cardol and MC), 1.65 - 1.55 (m, side chain), 1.40 - 1.28 (m, chain lateral), 0.89 - 0.86 (m, side chain). Traces of ethanol (< 1% by mass).

[0265] The mass percentages of the phenolic compounds of interest in natural CNSL extracted by solvent then hydrogenated are reported in the following table 13:

[0266] Table 13: Mass percentages of phenolic compounds in natural CNSL extracted by solvent then hydrogenated (sample A).

[0267] The melting temperature of the solvent-extracted hydrogenated natural CNSL was determined by DSC analysis using a DSC 200 F3 Maia® (Netzsch) heat flow instrument. 10 mg of the compound of interest was introduced into an aluminum crucible. A first temperature ramp from 20 to -25°C was applied, followed by an isotherm at -25°C for 15 minutes to stabilize the compound. Finally, a temperature ramp from -25 to 180°C was performed on the sample at a speed of 10 K / min, under nitrogen atmosphere (flow rate of 50 mL / min). The melting temperature of this CNSL was 88.5°C.

[0268] 10.3 - Natural CNSL extracted by press (sample C)

[0269] It is a natural CNSL extracted by pressing (mechanical extraction) at a sufficiently low temperature (< 150°C) to avoid decarboxylation of anacardic acid. The product was prepared by Orpia Innovation.

[0270] NMR- 1 H (400 MHz, DMSO-d6): 9.25 - 8.86 (wide signals, OH), 7.11 (1 H, t, J = 7.8 Hz, AA), 7.03 (1 H, t, J = 7.7 Hz, cardanol), 6.66 - 6.61 (2H, m, AA), 6.56 - 6.54 (3H, m, cardanol), 6.09 (2H, s, MC), 6.00 (3H, s, cardol), 5.84 - 5.74 (m, C=C), 5.42 - 5.27 (m, C=C), 5.05 - 4.94 (m, C=C), 2.81 - 2.72 (m, side chain), 2.65 - 2.61 (2H, m, AA), 2.49 -2.43 (2H, m, cardanol), 2.36 - 2.33 (2H, m, cardol and MC), 2.03 - 1.95 (m, side chain), 1.87 (3H, s, MC), 1.54 - 1.44 (m, side chain), 1.35 - 1.23 (m, side chain), 0.88 - 0.83 (m, side chain).

[0271] The mass percentages of the phenolic compounds of interest in natural CNSL extracted by press are reported in the following table 14:

[0272] Table 14: Mass percentages of phenolic compounds in natural CNSL extracted by press (sample C).

[0273] 10.4. - Natural CNSL extracted by press then hydrogenated (sample B)

[0274] 9.998 g of the natural CNSL extracted by press described previously in paragraph 10.3 were solubilized in 50 mL of ethanol at 30°C, then 0.508 g of Pd / C (5.1% by mass relative to the CNSL) were added to the medium. The suspension was then conditioned under an argon atmosphere, then under a dihydrogen atmosphere at 30°C for 4 days until the double bonds of the side chains had completely disappeared (the kinetic monitoring of the hydrogenation was carried out by NMR). 1H). The suspension was filtered under vacuum using a glass fiber filter paper, and the solid residue was washed on the frit with 2x10 mL of ethanol. The filtrate was again filtered under vacuum using a glass fiber paper, and then the solvent was removed using a rotary evaporator at 50°C. The solid was finally dried under high vacuum at room temperature for 18h30. Obtaining 9.005 g of natural CNSL extracted by press and then hydrogenated, i.e. a yield of 90.1% (with r = mcNSL h y dr °n ene y non-hydrogenated mCNSL

[0275] NMR- 1H (400 MHz, acetone-d6): 7.30 (1 H, t, J = 7.8 Hz, AA), 7.07 (1 H, t, J = 7.7 Hz, cardanol), 6.77 - 6.75 (2H, m, AA), 6.68 - 6.62 (3H, m, cardanol), 6.23 (2H, s, MC), 6.18 - 6.16 (3H, m, cardol), 3.00 (2H, broad signal, AA), 2.55 - 2.51 (2H, m, cardanol), 2.46 - 2.38 (2H, m, cardol and MC), 2.03 (3H, s, MC), 1.65 - 1.55 (m, side chain), 1.40 - 1 .28 (m, side chain), 0.89 - 0.86 (m, side chain). Traces of ethanol (< 1% by mass).

[0276] The mass percentages of the phenolic compounds of interest in natural CNSL extracted by press and then hydrogenated are reported in the following table 15: Table 15: Mass percentages of phenolic compounds in natural CNSL extracted by press then hydrogenated (sample B).

[0277] The melting temperature of the natural CNSL extracted by press and then hydrogenated was determined by DSC analysis using a DSC 200 F3 Maia® (Netzsch) heat flow instrument. 10 mg of compound of interest was introduced into an aluminum crucible. A first temperature ramp from 20 to -25°C was applied followed by an isotherm at -25°C for 15 minutes to stabilize the compound. Finally, a temperature ramp from -25 to 180°C was performed on the sample at a speed of 10 K / min, under nitrogen atmosphere (flow rate of 50 mL / min.). The melting temperature of this CNSL was 83.6°C.

[0278] 10.5. - CNSL technique (sample E)

[0279] It is a technical CNSL obtained by decarboxylation of a natural CNSL obtained by pressing.

[0280] NMR- 1H (400 MHz, CDCl3): 7.14 (1 H, t, J = 7.7 Hz, cardanol), 6.76 (1 H, t, J = 7.5 Hz, cardanol), 6.66 - 6.64 (2H, m, cardanol), 6.24 (broad signal, cardol and MC), 5.46 - 5.31 (m, C=C), 5.42 - 5.27 (m, C=C), 5.08 - 4.97 (m, C=C), 2.83 - 2.78 (m, side chain), 2.55 (2H, t, J = 7.7 Hz, cardanol), 2.50 - 2.45 (2H, very broad signal, cardol and MC), 2.10 (3H, s, MC), 2.07 - 1.99 (m, chain side chain), 1.63 - 1.56 (m, side chain), 1.40 - 1.26 (m, side chain), 0.91 - 0.87 (m, side chain).

[0281] The product obtained comprises by weight approximately 82% cardonol and 18% cardol and traces of methyl cardol, according to NMR spectrum analysis.

[0282] 10.6. - Hydrogenated technical CNSL (sample F)

[0283] 15.152 g of the technical CNSL described previously in paragraph 10.5 were solubilized in 75 mL of ethanol at 30°C, then 1.049 g of Pd / C (6.92% by mass relative to the CNSL) were added to the medium. The suspension was then conditioned under argon atmosphere, then under dihydrogen atmosphere at 30°C for 7 days. The suspension was filtered under vacuum using a glass fiber filter paper and the solid residue was washed on the frit with 2x20 mL of ethanol. The filtrate was again filtered under vacuum using a glass fiber paper and then the solvent was removed using a rotary evaporator at 50°C. NMR analysis of this compound indicated that it still contained unsaturations in the side chains of the phenolic compounds. Therefore, it was decided to restart the hydrogenation of the remaining double bonds.The oily residue was solubilized in 80 mL of ethanol at 30°C before adding 752 mg of Pd / C, then the suspension was conditioned under an argon atmosphere and then dihydrogen for 2 days. The suspension was then filtered according to the protocol described previously. The solvent from the filtrate was removed using a rotary evaporator at 50°C, and the solid thus obtained was dried under high vacuum at room temperature for 14 h. Obtaining 13.252 g of a light brown solid of hydrogenated technical CNSL, i.e. a yield of 87.5% (with r =. mcNSL h y dr °n ene y non-hydrogenated mCNSL

[0284] NMR- 1H (400 MHz, acetone-d6): 8.14 (broad signal, OH), 7.07 (1 H, t, J = 7.7 Hz, cardanol), 6.68 - 6.62 (3H, m, cardanol), 6.23 (2H, s, MC), 6.19 - 6.16 (3H, m, cardol), 3.00 (2H, broad signal, AA), 2.54 - 2.51 (2H, m, cardanol), 2.44 - 2.37 (2H, m, cardol and MC), 2.03 (3H, s, MC), 1.62 - 1.55 (m, side chain), 1.35 - 1.27 (m, side chain), 0.89 - 0.86 (m, side chain). Traces of ethanol.

[0285] The mass percentages of the phenolic compounds of interest in the hydrogenated technical CNSL are reported in the following table 16:

[0286] Table 16: Mass percentages of phenolic compounds in hydrogenated technical CNSL (sample F).

[0287] The melting temperature of the hydrogenated technical CNSL was determined by DSC analysis using a DSC 200 F3 Maia® (Netzsch) heat flow instrument. 10 mg of the compound of interest was introduced into an aluminum crucible. A first temperature ramp from 20 to -25°C was applied, followed by an isotherm at -25°C for 15 minutes to stabilize the compound. Finally, a temperature ramp from -25 to 180°C was performed on the sample at a speed of 10 K / min, under nitrogen atmosphere (flow rate of 50 mL / min). The melting temperature of the hydrogenated technical CNSL was 46.9°C.

[0288] Example 11 - In vitro evaluation of the anticoccidial effect - Materials and methods

[0289] The in vitro models developed allow rapid evaluation of the anticoccidial effect of a product under several conditions, without the need to use live chickens. The effects of samples A to F on sporulation, viability, and invasion capacity of Eimeria parasites were evaluated at eight doses ranging from 6.25 to 800 pg / mL. Two anticoccidials (50 pg / mL monensin and 50 pg / mL amprolium) were used as positive controls, in addition to a dead or sporulated control and an untreated live or sporulated control.

[0290] 11.1. Products tested and preparation of solutions for biological tests

[0291] The nature of the products, the name and the reference corresponding to the 6 tested products prepared according to example 10 above, namely samples A, B, C, D, E and F are reported in table 17.

[0292] Table 17: Products tested.

[0293] The preparation of solutions of the products tested for biological tests is as follows:

[0294] 1. Preparation of a stock solution of each tested product at 200,000 pg / mL in DMSO (4g of tested product in a volume of 20 mL).

[0295] 2. Preparation of a 20,000 pg / mL solution in the incubation buffer solution (PBS, potassium dichromate or culture medium depending on the test performed) from the stock solution.

[0296] 3. Preparation of serial dilutions (1 / 2) in incubation buffer from the 20,000 pg / mL solution in a range of test product doses between 6.25 and 800 pg / mL, including doses of 6.25 pg / mL, 12.5 pg / mL, 25 pg / mL, 50 pg / mL, 100 pg / mL, 200 pg / mL, 400 pg / mL and 800 pg / mL were prepared.

[0297] 11.2. Sporulation of oocysts

[0298] Oocyst sporulation rates were determined in vitro according to the procedures described in Fatemi et al. (Parasitology Research 114, 1207-1211, 2015). Thus, 1 x 10 5 Mixed Eimeria oocysts were incubated with 8 doses (from 6.25 pg / mL to 800 pg / mL) of each sample A to F at 29°C in 2% potassium dichromate with aeration for 48 h. The numbers of sporulated and non-sporulated oocysts were measured by the hemocytometer method (Conway and McKenzie, 2007). At least 100 oocysts were assessed per replicate. Three replicates per condition were used.

[0299] Several witnesses were included in this study:

[0300] A positive control consisted of amprolium (50 pg / mL) an anticoccidial agent.

[0301] A second positive control consisted of monensin (50 pg / mL), another anticoccidial agent.

[0302] A control consisting of non-sporulated oocysts.

[0303] A control consisting of untreated sporulated oocysts.

[0304] 11.3. Sporozoite viability

[0305] Viability percentages were determined in vitro according to the procedures described in Kim et al. (The British Journal of Nutrition 109, 76-88, 2013). Briefly, field Eimeria oocysts were lysed to release sporozoites. 1 x 10 6 Sporozoites were incubated with 8 doses (from 6.25 pg / mL to 800 pg / mL) of each sample A to F (culture medium: PBS) at 4°C for 2 h, and then viability was measured by trypan blue exclusion and motility by counting at least 100 sporozoites per condition. Three replicates per condition were used.

[0306] Several witnesses were included in this study:

[0307] A positive control consisted of amprolium (50 pg / mL) an anticoccidial agent.

[0308] A second positive control consisted of monensin (50 pg / mL), another anticoccidial agent.

[0309] A witness consisting of dead sporozoites.

[0310] A control consisting of untreated live sporozoites. 11.4. MDBK host cell invasion assays by Eimeria species

[0311] 11.4. 1. Cytotoxicity tests on MDBK cells

[0312] According to methods adapted from Kabir et al. (PLOS Neglected Trop. Dis. 16, e0010947, 2022), Madin-Darby Bovine Kidney (MDBK) cells, cultured in 96-well plates for 24 h at a rate of 4 x 10 4Cells per well were treated with the same eight doses (from 6.25 pg / mL to 800 pg / mL) of each sample A to F for 48 h, then equilibrated at room temperature (20 to 30°C) for 30 min. Cell viability was then determined by Cell-Titer Gio reagent (Promega, Madison, WI) according to the manufacturer's instructions. The luminescence signal of control wells containing 0.1% DMSO was set at 100% cell viability.

[0313] 11.3.2. Sporozoite invasion capacity

[0314] Sporozoite invasion rates were determined in vitro according to the procedures described in Burt et al. (Veterinary Parasitology 191, 374-378, 2013). Thus, MDBK bovine kidney epithelial cells at 70-80% confluence were contacted with samples A to F and then exposed to 2 x 10 5sporozoites and incubated at 37°C with 5% CO2 for 2 h, 4 h and 20 h. The cells were then washed and collected. Their DNA was extracted and qPCR was performed to evaluate the differential effect on Eimeria species, namely E. acervulina, E. maxima and E. tenella, present in the field strain isolate collected by the laboratory in 2018. A control treatment, without sporozoites, was also used. Three replicates per condition were used.

[0315] 11.5. In vitro test analyzes

[0316] Half-maximal inhibitory concentrations (EC50) and half-maximal cellular cytotoxic concentrations (CC50) of each test product were calculated by nonlinear regression sigmoidal dose-response curve fitting (GraphPad Prism, version 10). Differences in parasitemia between treatments and control were considered significant with p < 0.05 using one-way variance analysis, followed by Tukey post-hoc tests. For all in vitro assays, three replicates per condition were used.

[0317] References Burt, SA, Tersteeg-Zijderveld, Jongerius-Gortemaker, BGM, Vervelde, L, Vernooij,

[0318] 2013. In vitro inhibition of Eimeria tenella invasion of epithelial cells by phytochemicals.

[0319] Veterinary parasitology 191, 374-378. doi.org / 10.1016 / j.vetpar.2012.09.001

[0320] Conway, D.P., McKenzie, M.E., 2007. Poultry Coccidiosis, John Wiley & Sons. John Wiley & Sons, https: / / doi.org / 10.1002 / 9780470344620 Fatemi, A., Razavi, S.M., Asasi, K., Goudarzi, M.T., 2015. Effects of Artemisia annua extracts on sporulation of Eimeria oocysts. Parasitology research 114, 1207-1211. doi.org / 10.1007 / s00436- 014-4304-z

[0321] Kabir, M.H.B., Recuenco, F.C., Zin, N.K.M., Watanabe, N., Fukuda, Y, Bando, H., Watanabe, K., Bochimoto, H., Xuan, X., Kato, K., 2022. Identification of potent anti-Cryptosporidium new drug leads by screening traditional Chinese medicines. PLOS Neglected Trop. Dis. 16, e0010947. doi.org / 10.1371 / journal.pntd.0010947

[0322] Kim, D.K., Lillehoj, H.S., Lee, S.H., Lillehoj, E.P, Bravo, D., 2013. Improved resistance to Eimeria acervulina infection in chickens due to dietary supplementation with garlic metabolites. The British journal of nutrition 109, 76-88. doi.org / 10.1017 / s0007114512000530

[0323] Example 12: Results

[0324] Sporozoite viability

[0325] The results of the sporozoite viability tests and the half-maximal inhibitory concentration (ECso) analysis are presented respectively: in Figure 11 for the hydrogenated natural CNSL obtained by solvent (Product A), in Figure 12 for the hydrogenated natural CNSL obtained by press (Product B), in Figure 13 for the natural CNSL obtained by press (Product C), in Figure 14 for the natural CNSL obtained by solvent (Product D), in Figure 15 for the technical CNSL (Product E). in Figure 16 for the hydrogenated technical CNSL (Product F).

[0326] The half-maximal inhibitory concentration (ECso) values ​​of the tested products are presented in the following Table 18.

[0327] Table 18: Maximum inhibitory concentration (EC50) values.

[0328] It is observed that: hydrogenated natural CNSL exhibits more effective activity against the pathogen, sporozoites, than natural, i.e. non-hydrogenated CNSL and that technical CNSL, natural CNSL is more effective than technical CNSL. hydrogenated technical CNSL is as effective as hydrogenated natural CNSL.

[0329] Cytotoxicity on MDBK cells and invasion of E. tenella sporozoites Regarding the MDBK host cell invasion assays, an anticoccidial effect of hydrogenated CNSL is observed at a concentration well below the half-maximal cellular cytotoxic concentration (CC50) on MDBK cells, as reported in Figures 17 and 18. The half-maximal cellular cytotoxic concentration (CC50) data of MDBK cells exposed to samples A to F, as well as the half-maximal inhibitory concentration (EC50) values ​​for E. tenella sporozoite invasion for each sample A to F, are reported in Table 19. Table 19: Half-maximal cellular cytotoxic concentration (CC50) values ​​of MDBK cells exposed to samples A to F and half-maximal inhibitory concentration (EC50) values ​​of invasion of E. tenella sporozoites exposed to the same samples. All tested products perform better than the amprolium reference, which has no effect against the E. tenella strain, or even promotes its proliferation. The tested pathogen appears to be resistant to amprolium.

[0330] All products are as effective as, or even more effective than, the monensin reference (tested at 50 micrograms / mL) at all tested concentrations. Regarding the comparison of the tested products: Technical CNSL (Product E) is the least effective among the 6 products tested. Natural CNSL and hydrogenated natural CNSL have comparable effectiveness (EC50s are of the same order of magnitude). Hydrogenated technical CNSL (Product F) works as well as, or even better than, hydrogenated natural CNSL.

Claims

CLAIMS 1. Non-therapeutic use of a composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV): wherein said composition comprises: - hydrogenated anacardic acid from 50.0 to 85.0% by total weight of the composition, - hydrogenated cardol from 10.0 to 25.0% by total weight of the composition - hydrogenated methyl cardol from 0.0 to 5.0% by total weight of the composition, and - hydrogenated cardanol from 2.0 to 15.0% by total weight of the composition, or non-therapeutic use of hydrogenated anacardic acid of formula (I) to reduce methanogenesis and / or to improve digestive fermentation in animals.

2. Use according to claim 1, wherein said composition is derived from a cashew nut extract obtained by extraction using a solvent or mechanically without heating.

3. Use according to one of claims 1 to 2, wherein said composition of hydrogenated cashew nut shell extract or hydrogenated anacardic acid is used at a dose of 50 to 600 ppm, in particular 50 to 500 ppm.

4. Use according to one of claims 1 to 3, for reducing methanogenesis and possibly improving digestive fermentation in the rumen of ruminants and / or possibly for increasing propionate production, in particular of cattle.

5. Association of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV) following: in which - hydrogenated anacardic acid represents from 50.0 to 85.0% by total weight of the association, - hydrogenated cardol represents from 10.0 to 25.0% by total weight of the association, - hydrogenated methyl cardol represents from 0.0 to 5.0% by total weight of the association, and - hydrogenated cardanol represents from 2.0 to 15.0% by total weight of the association.

6. Association of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV) following: in which - hydrogenated anacardic acid represents from 0.0 to 30.0% by total weight of the association, - hydrogenated cardol represents from 10.0 to 25.0% by total weight of the association, - hydrogenated methyl cardol represents from 0.0 to 5.0% by total weight of the association, and - hydrogenated cardanol represents from 50.0 to 85.0% by total weight of the association.

7. Use of a combination according to claim 5 or of hydrogenated anacardic acid as a food supplement for reducing methanogenesis and / or for improving digestive fermentation, in particular as a food supplement for ruminants.

8. Use of a combination according to claim 5 or of hydrogenated anacardic acid for the manufacture of a food supplement for reducing methanogenesis and / or for improving digestive fermentation.

9. Use according to one of claims 7 or 8, said association or anacardic acid being used at a rate of 50 to 600 ppm, in particular 50 to 500 ppm.

10. Food comprising the combination according to claim 5 or hydrogenated anacardic acid, at a rate of 50 to 600 ppm, in particular 50 to 500 ppm.

11. Hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract composition comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV): in which - hydrogenated anacardic acid represents from 50.0 to 85.0% by total weight of the association, - hydrogenated cardol represents from 10.0 to 25.0% by total weight of the association, - hydrogenated methyl cardol represents from 0.0 to 5.0% by total weight of the association, and - hydrogenated cardanol represents from 2.0 to 15.0% by total weight of the association, or composition comprising or consisting of hydrogenated anacardic acid, for its use against digestive pathological conditions chosen from abdominal tympanism or coccidiosis.

12. Composition for use according to claim 11, against abdominal tympanism.

13. Composition for use according to claim 11, against coccidiosis.

14. Composition for use according to claim 13, said composition being at a dose of 5.00 to 1000 μg by weight of said composition in 1 mL of a pharmaceutically acceptable medium.

15. Composition of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract for use against coccidiosis.

16. Composition for use according to claim 15 of hydrogenated cashew nut shell liquid (hydrogenated CNSL) or hydrogenated cashew nut shell extract comprising or consisting of the following combination of hydrogenated anacardic acid of formula (I), hydrogenated cardol of formula (II), hydrogenated methyl cardol of formula (III) and hydrogenated cardanol of formula (IV): in which - hydrogenated anacardic acid represents from 0.0 to 30.0% by total weight of the association, - hydrogenated cardol represents from 10.0 to 25.0% by total weight of the association, - hydrogenated methyl cardol represents from 0.0 to 5.0% by total weight of the association, and - hydrogenated cardanol represents from 50.0 to 85.0% by total weight of the association for its use against coccidiosis.

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