Methane production inhibitors in ruminants

Abstract

Provided is a methane production inhibitor for ruminants, which is characterized by containing cashew nut shell oil, heat-treated cashew nut shell oil, anacardic acid, cardanol and / or cardol, and an organic acid as active ingredients, the mass ratio of the matching amount of the cashew nut shell oil, the cashew nut shell oil subjected to heat treatment, the anacardic acid, the cardanol and / or the cardol to the matching amount of the organic acid is (1: 5)-(1: 20).

Description

Technical Field

[0001] The present invention relates to a preparation and a feed for suppressing methane production in ruminants such as cattle, and a method for suppressing methane production in ruminants using the same. Background Art

[0002] Ruminants such as cattle and sheep digest and ferment feed with the help of microorganisms in the rumen and survive by using the fermentation products. On the other hand, the production of methane by methanogenic bacteria in the rumen causes a loss in the energy efficiency of the feed. In addition, methane is a greenhouse gas that affects global warming, so it is important to reduce the production of methane in the rumen of ruminants.

[0003] Methanogens in the rumen use hydrogen to reduce carbon dioxide to produce methane. Methane is the second largest contributor to global warming after carbon dioxide, and ruminants are thought to produce 15-20% of total methane emissions.

[0004] Cashew nut shell liquid is known to have an effect on ruminants, including an effect of inhibiting methane production, to improve rumen fermentation (Patent Document 1: WO2008 / 149992). However, due to the increasing global awareness of preventing global warming in recent years, further improvements are being sought.

[0005] On the other hand, Non-Patent Document 1 (Animal Feed Science and Technology, Volumes 166-167, 23 June 2011, Pages 282-290) and Non-Patent Document 2 (Journal of Dairy Science, Volume 92, Issue 7, July 2009, Pages 3258-3264) disclose that malic acid inhibits methane production, but this is not practical because very high concentrations of malic acid must be used.

[0006] In addition, Patent Document 2 (WO2011 / 152532) and Patent Document 3 (WO2012 / 060451) disclose that an acid such as an organic acid is added to a composition containing cashew nut shell liquid in order to suppress the decomposition of anacardic acid and improve stability. However, the organic acid is added in a trace amount relative to the cashew nut shell liquid, and there is no suggestion of suppressing the generation of methane.

[0007] Prior art literature

[0008] Patent Literature

[0009] Patent Document 1: WO2008 / 149992

[0010] Patent Document 2: WO2011 / 152532

[0011] Patent Document 3: WO2012 / 060451

[0012] Non-patent literature

[0013] Non-patent literature 1: Animal Feed Science and Technology, Volumes 166-167, 23 June 2011, Pages 282-290

[0014] Non-patent document 2: Journal of Dairy Science, Volume 92, Issue 7, July 2009, Pages 3258-3264 Summary of the invention

[0015] Problems to be solved by the invention

[0016] An object of the present invention is to provide a material for effectively suppressing methane production while maintaining energy production and metabolism in ruminants.

[0017] Means for solving problems

[0018] The present inventors have conducted intensive research to solve the above problems and have found that by administering cashew nut shell liquid (hereinafter sometimes referred to as CNSL) and an organic acid to ruminants at a specific ratio, the production of acetic acid, which is the main energy and lipid synthesis material of ruminants, can be maintained while effectively suppressing the production of methane in ruminants. Based on this finding, the present inventors have completed the present invention.

[0019] That is, the present invention is as follows.

[0020] [1] A methane production inhibitor for ruminants, characterized in that it contains cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol, and an organic acid as active ingredients, and the mass ratio of the cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol to the mass ratio of the organic acid to the mass ratio is 1:5 to 1:20 (or 1:10 to 1:20).

[0021] [2] The methane generation inhibitor according to [1], wherein the organic acid is a carboxylic acid having 3 to 12 carbon atoms.

[0022] [3] The methane generation inhibitor according to [1], wherein the organic acid is a hydroxycarboxylic acid having 3 to 10 carbon atoms.

[0023] [4] The methane production inhibitor according to [1], wherein the organic acid is citric acid or malic acid.

[0024] [5] The methane production inhibitor according to any one of [1] to [3], wherein the ruminant is a cattle.

[0025] [6] A feed for inhibiting methane production in ruminants, comprising the methane production inhibitor according to any one of [1] to [5].

[0026] [7] A method for inhibiting methane production in ruminants, characterized in that the method includes a step of administering cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and an organic acid to the ruminants, wherein the mass ratio of the amount of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol administered per ruminant per day to the amount of organic acid administered per ruminant per day is 1:5 to 1:20 (or may be 1:10 to 1:20).

[0027] [8] The method according to [7], wherein the organic acid is a carboxylic acid having 3 to 12 carbon atoms.

[0028] [9] The method according to [7], wherein the organic acid is a hydroxycarboxylic acid having 3 to 10 carbon atoms.

[0029]

[10] The method according to [7], wherein the organic acid is citric acid or malic acid.

[0030]

[11] The method according to any one of [7] to

[10] , wherein the ruminant is a cattle.

[0031]

[12] The method according to any one of [7] to

[11] , wherein the cashew nut shell liquid, the heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and the organic acid are mixed in advance before application.

[0032]

[13] The method according to any one of [7] to

[11] , wherein cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and an organic acid are applied separately.

[0033]

[14] An application, characterized in that it is the use of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and an organic acid in the manufacture of a methane production inhibitor for ruminants, wherein the mass ratio of the cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol to the mass ratio of the organic acid is 1:5 to 1:20 (or may be 1:10 to 1:20).

[0034]

[15] The use according to

[14] , wherein the organic acid is a carboxylic acid having 3 to 12 carbon atoms.

[0035]

[16] The use according to

[14] , wherein the organic acid is a hydroxycarboxylic acid having 3 to 10 carbon atoms.

[0036]

[17] The use according to

[14] , wherein the organic acid is citric acid or malic acid.

[0037]

[18] The use according to any one of

[14] to

[17] , wherein the ruminant is a cattle.

[0038]

[19] An application, characterized in that it is a non-therapeutic application of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and an organic acid for inhibiting methane production in ruminants, and the mass ratio of the cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol to the mass ratio of the organic acid is 1:5 to 1:20 (or 1:10 to 1:20).

[0039]

[20] The use according to

[19] , wherein the organic acid is a carboxylic acid having 3 to 12 carbon atoms.

[0040]

[21] The use according to

[19] , wherein the organic acid is a hydroxycarboxylic acid having 3 to 10 carbon atoms.

[0041]

[22] The use according to

[19] , wherein the organic acid is citric acid or malic acid.

[0042]

[23] The use according to any one of

[19] to

[22] , wherein the ruminant is a cattle.

[0043] Effects of the Invention

[0044] According to the present invention, by administering cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol, and / or cardanol and organic acid to ruminants at a specific ratio, methane production in the rumen can be effectively suppressed while maintaining the production of acetic acid, which is the main energy and lipid synthesis material of ruminants. Thus, it is possible to contribute to the prevention of global warming in the livestock field while maintaining the growth and productivity of livestock. Cashew nut shell liquid works under low dosage conditions, so it has the advantage of high safety. DETAILED DESCRIPTION

[0045] The methane generation inhibitor of the present invention is characterized in that it contains cashew nut shell liquid (CNSL), heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol (sometimes referred to as cashew nut shell liquid, etc. in this specification) and an organic acid as active ingredients, and the mass ratio of the amount of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol to the amount of the organic acid is 1:5 to 1:20 (or 1:10 to 1:20).

[0046] Here, the amount of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol blended is the total blended amount when a plurality of these substances are contained, and the amount of organic acid blended is the total blended amount when two or more organic acids are contained.

[0047] When cashew nut shell liquid is described separately from heat-treated cashew nut shell liquid, it means non-heated cashew nut shell liquid.

[0048] Among cashew nut shell liquid (non-heated), heated cashew nut shell liquid, anacardic acid and cardanol, cashew nut shell liquid (non-heated) or anacardic acid is preferred, and cashew nut shell liquid (non-heated) is particularly preferred.

[0049] Cashew nut shell liquid is an oily liquid contained in the shell of the fruit of the cashew tree (Anacardium occidentale L.). Cashew nut shell liquid contains anacardic acid, cardanol, and cardanol as its components. Anacardic acid is converted into cardanol by heating, and cashew nut shell liquid (heated cashew nut shell liquid) that is converted into only cardanol and cardanol by heating can also be used.

[0050] Cashew nut shell liquid (non-heated) extracted by low-temperature pressing of cashew nut shells contains 55 to 80% by mass of anacardic acid, 5 to 20% by mass of cardanol, and 5 to 30% by mass of cardanol as described in J. Agric. Food Chem. 2001, 49, 2548-2551.

[0051] The heated cashew nut shell liquid obtained by heating the non-heated cashew nut shell liquid (for example, at 70° C. or higher, preferably at 130° C. or higher) contains 0 to 10% by mass of anacardic acid, 55 to 80% by mass of cardanol, and 5 to 30% by mass of cardanol due to decarboxylation of anacardic acid, which is a main component of the non-heated cashew nut shell liquid.

[0052] Cashew nut shell liquid can be obtained as a vegetable oil extracted by low temperature pressing of cashew nut shells. Alternatively, cashew nut shell liquid can be obtained by dry distillation or solvent extraction of cashew nut shells. Alternatively, cashew nut shell liquid can be obtained by the method described in Japanese Patent Application Laid-Open No. 8-231410. Commercially available cashew nut shell liquid can also be used.

[0053] The methane production inhibitor of the present invention may contain one or more of anacardic acid, cardanol, and cardanol instead of cashew nut shell liquid.

[0054] As anacardic acid, natural anacardic acid, synthetic anacardic acid, and their derivatives can be cited. In addition, commercially available anacardic acid can also be used. Anacardic acid can be obtained by the operation described in Japanese Patent Laid-Open No. 8-231410, that is, the shell of cashew nut is subjected to extraction treatment with an organic solvent, and the obtained cashew nut oil is eluted by, for example, using silica gel column chromatography to change the ratio of a mixed solvent of n-hexane, ethyl acetate and acetic acid (Japanese Patent Laid-Open No. 3-240721, Japanese Patent Laid-Open No. 3-240716, etc.).

[0055] Examples of cardanol include natural cardanol, synthetic cardanol, and derivatives thereof. The cardanol used in the present invention can be obtained by decarboxylating anacardic acid, which is a main component of cashew nut shell liquid.

[0056] Examples of cardanol include natural cardanol, synthetic cardanol, and derivatives thereof. In addition, the cardanol used in the present invention can be obtained by purifying cashew nut shell liquid.

[0057] The content of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol in the methane generation inhibitor of the present invention (the total amount when two or more types are contained) is, for example, 1% to 20% by mass based on the total amount of the preparation.

[0058] The type of organic acid is not particularly limited, but is preferably a carboxylic acid, and the carboxylic acid includes monocarboxylic acid, dicarboxylic acid, and tricarboxylic acid, and examples thereof include aliphatic carboxylic acid, aliphatic hydroxycarboxylic acid, aromatic carboxylic acid, aromatic hydroxycarboxylic acid, and amino acid. The number of carbon atoms in the organic acid is preferably 3 or more, more preferably 4 or more, including the carbon atoms in the carboxyl group, and on the other hand, is preferably 12 or less, more preferably 10 or less, further preferably 8 or less, and particularly preferably 7 or less. Specific examples of organic acids include formic acid, acetic acid, propionic acid, butyric acid, valeric acid, salicylic acid, lactic acid, tartaric acid, citric acid, succinic acid, malic acid, fumaric acid, ricinoleic acid, quinic acid, and shikimic acid, but citric acid, malic acid, or fumaric acid is particularly preferred.

[0059] The content of the organic acid in the organic acid methane generation inhibitor (the total amount when two or more kinds are included) is, for example, 5% by mass to 95% by mass based on the total amount of the preparation.

[0060] The methane production inhibitory effect of the methane production inhibitor of the present invention can be ascertained, for example, by comparing the methane production amount when CNSL, etc. and an organic acid are added to a culture medium of rumen fluid of a ruminant at the above-mentioned specific ratio and cultured with the production amount of a control (when CNSL, etc. and an organic acid are not added) as described in the examples below. It should be noted that CNSL, etc. and an organic acid can also be directly administered to ruminants to compare methane production. The effect of CNSL, etc. and an organic acid is preferably to reduce the methane production amount to 90% or less, preferably to 80% or less, and more preferably to 50% or less compared to the control (when CNSL, etc. and an organic acid are not added).

[0061] The methane production inhibitor of the present invention may contain, in addition to cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and organic acid, ingredients such as excipients that can be used in feeds or medicines, such as lactose, white sugar, D-mannitol, α-starch, starch, corn starch, crystalline cellulose, bentonite, silica gel, and light silicic anhydride.

[0062] In addition, the methane production inhibitor of the present invention may further contain, in addition to cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and organic acid, an arbitrary component such as a component effective for promoting the growth of ruminants, a nutritional supplement component, a component for improving storage stability, a coating agent component, etc. Such arbitrary components include raw materials for feed such as wheat bran, alfalfa, and timothy hay, feed additives, food raw materials, food additives, pharmaceutical raw materials, and other supplement components used in animal supplements (hereinafter referred to as supplements). For example, there can be mentioned live bacteria such as enterococci, Bacillus, and bifidobacteria; enzymes such as amylase and lipase; L-ascorbic acid, choline chloride, inositol, and folic acid; minerals such as potassium chloride, magnesium oxide, and phosphates; antioxidants such as ethoxyquin, butylated hydroxytoluene, butylated hydroxyanisole, ferulic acid, vitamin C, and vitamin E; mold inhibitors such as calcium propionate; binders such as carboxymethyl cellulose (CMC), sodium caseinate, and sodium polyacrylate; emulsifiers such as lecithin, fatty acid glycerides, and sorbitan fatty acid esters; pigments such as astaxanthin and cantharidin; and flavoring agents such as various esters, ethers, and ketones.

[0063] The methane generation inhibitor of the present invention may contain an oil absorbent such as magnesium oxide, stearate, talc, zeolite, diatomaceous earth, and silica, and the oil absorbent is preferably in a particle form. As the oil absorbent, an oil absorbent that absorbs 50 to 300 g of oil per 100 g is preferred. In addition, if the particle size is greater than 300 μm, the particles become coarse and gradually separate, so particles with a particle size of 2 to 300 μm are preferred.

[0064] In one embodiment of the methane production inhibitor of the present invention, a preferred mass ratio of the oil absorbing agent to the cashew nut shell liquid, the heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol, and the organic acid is 100:20 to 100:180.

[0065] The dosage form of the methane generation inhibitor of the present invention is not particularly limited, and may be any form such as liquid, powder, solid, tablet, capsule, emulsion, granule, coated agent, etc., but liquid, powder, capsule, granule, tablet is preferred.

[0066] As a liquid preparation, cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol can be used directly and combined with an organic acid, or cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol can be dissolved in a solvent such as ethanol and combined with an organic acid, or the above-mentioned excipients or optional components can be further added before use. In addition, the following powders, capsules, granules, and tablets can also be suspended or floated in a liquid.

[0067] As a powder, the above-mentioned excipients may be added to cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diol, and an organic acid and the mixture may be powdered.

[0068] As capsules, cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diol, and organic acid may be directly filled into capsules, or the above-mentioned excipients or optional ingredients may be added.

[0069] The granules may be prepared by adding the above excipients to cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diol, and an organic acid, followed by granulation.

[0070] Tablets may be prepared by adding the above excipients to cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diol, and an organic acid, followed by granulation to obtain tablets.

[0071] When an oil absorbent such as silica is contained, it is preferably formulated in the form of a powder, tablet or granule.

[0072] As described above, the methane generation inhibitor of the present invention can be prepared by mixing cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol with an organic acid, and then mixing an excipient or an optional component as needed, and preparing a formulation. It should be noted that, depending on the form of the formulation, the above-mentioned cashew nut shell crushed / crushed material, or the cashew nut shell without any treatment can be directly mixed with the organic acid and other optional components to form the methane generation inhibitor of the present invention.

[0073] Cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol are preferably mixed with an organic acid and then administered to an animal feed.

[0074] Therefore, the present invention provides a feed for suppressing methane production in ruminants, comprising cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diol, and an organic acid as active ingredients.

[0075] The feed of the present invention is characterized by containing cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol, and an organic acid.

[0076] The content of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol (the total amount when two or more types are included) in the feed of the present invention is not particularly limited as long as the mass ratio of the amount of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol to the amount of organic acid is 1:5 to 1:20 (or 1:10 to 1:20).

[0077] The feed of the present invention can be produced by adding cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol, and an organic acid or the methane production inhibitor of the present invention containing them to feed ingredients and mixing them.

[0078] At this time, in the case of using powdered or solid preparations, liquid carriers can be used to make the preparations into liquid or gel forms in order to make mixing easy. In this case, fluid liquids such as water, vegetable oil, liquid animal oil, mineral oil, synthetic oil, and water-soluble polymer compounds can be used as liquid carriers. In addition, in order to ensure the uniformity of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and organic acid in feed, it is also preferred to coordinate water-soluble polysaccharides such as alginic acid, sodium alginate, xanthan gum, carboxymethyl cellulose, alpha starch, sodium caseinate, gum arabic, guar gum, and tamarind seed polysaccharides.

[0079] There are no particular restrictions on the type and proportion of the feed ingredients to be combined with the preparation of the present invention in the feed of the present invention, as long as they are feeds that have been provided to various animals in the past. For example, corn kernels, corn flour, millet, wheat bran, soybean meal, oats, wheat low-gluten flour, wheat meal, alfalfa, timothy hay, clover, defatted rice bran, northern fish meal, coastal fish meal, yeast, molasses, meat slices, bone meal, calcium carbonate, calcium hydrogen phosphate, yellow grease, vitamins, minerals, etc. can be used for preparation.

[0080] In addition, the present invention provides a method for inhibiting methane production in ruminants, characterized in that it includes a step of administering cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and an organic acid to the ruminants, wherein the mass ratio of the amount of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol administered per ruminant per day to the amount of organic acid administered per ruminant per day is 1:5 to 1:20 (or may be 1:10 to 1:20).

[0081] Cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and organic acid can be administered to feeds containing them, i.e., they can be mixed in advance and then administered, or they can be administered separately to ruminants. In the case of separate administration (feeding), the timing of administration is not particularly limited as long as they are ingested by ruminants and exist in the body, especially in the rumen.

[0082] The type of ruminant that ingests the feed of the present invention is not particularly limited, and examples thereof include cattle, buffalo, goat, sheep, yak, etc. Here, examples of cattle species include Holstein, Jersey, Kuroko, Japanese Shorthorn, and Aberdeen Angus, but are not limited thereto. In addition, the use of livestock products such as dairy and meat products for production and breeding is not particularly limited. The period of provision of this agent can be exemplified by use after 4 weeks of age when the rumen begins to form functions, but is not limited thereto.

[0083] The amount of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol (the total amount when two or more are included) administered per ruminant per day may be, for example, 0.5 to 50 g / head / day, preferably 1 to 25 g / head / day, and more preferably 2.5 to 10 g / head / day. The amount of organic acid (the total amount when two or more are included) administered per ruminant per day may be, for example, 1 to 1000 g / head / day, preferably 5 to 200 g / head / day, and more preferably 12.5 to 150 g / head / day. The mass ratio of the daily administration amount of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol per ruminant to the daily administration amount of the organic acid per ruminant is adjusted to 1:5 to 1:20 (or 1:10 to 1:20).

[0084] The administration may be performed multiple times a day or over several days.

[0085] Example

[0086] Hereinafter, the present invention will be specifically described with reference to Examples, but the embodiments of the present invention are not limited to the following Examples.

[0087] Example 1

[0088] Evaluation of cashew nut shell liquid (CNSL), organic acids, and minced garlic in a methane production measurement system using rumen culture fluid

[0089] (1) Sample

[0090] Cashew nut shell liquid (CNSL) extracted from cashew nut shells by low temperature pressing was obtained. Rumen fluid (four-layer gauze filtrate) collected orally from Holstein lactating cows more than 90 days after parturition was used as the culture inoculum. 4 mL of the rumen fluid and 6 mL of artificial saliva (McDougal's artificial saliva) were added for culture.

[0091] (2) Cultivation

[0092] The concentration of CNSL in the test culture medium was set to 100-500 mg / L. CNSL was dissolved in ethanol and added to a Hungate tube to an arbitrary concentration in the solution. DL-malic acid and fumaric acid were similarly dissolved in ethanol and added to an arbitrary concentration in the solution. In order to make the effect of ethanol equal, only ethanol was added to the non-added area (control area). After that, the ethanol was allowed to volatilize by leaving it for several hours, and 50 mg of corn starch, 75 mg of rich feed powder, and 75 mg of timothy hay powder were added as a culture matrix. Garlic powder was added at 0.5% or 1.0% relative to the dry matter of the culture matrix. This was equivalent to 100 mg / L or 200 mg / L when converted to the concentration in the test culture medium. A total of 10 ml of diluted rumen fluid and artificial saliva were added to these test tubes at the above mixing ratio, and butyl rubber caps and plastic screw caps were covered while nitrogen was blown into the headspace, and anaerobic culture was carried out in an incubator (39°C, 17 hours). Each of the three cultures was cultured in triplicate.

[0093] (3) Analysis

[0094] The concentrations of methane and acetic acid in volatile fatty acids were analyzed using FID gas chromatography.

[0095] (4) Results

[0096] (i) Gas production and acetic acid production

[0097] Table 1 shows the results of the gas generation amount and the acetic acid generation amount.

[0098] A decrease in methane production and acetic acid production was observed depending on the CNSL concentration. When 100 mg / L CNSL and 500-1000 mg / L DL-malic acid or 500 mg / L fumaric acid were used in combination, methane production was significantly reduced compared to the addition of 100 mg / L CNSL alone, while no significant decrease in acetic acid production was observed. No significant increase in methane reduction was observed under the conditions of the combination of CNSL and minced garlic, the use of 500-1000 mg / L DL-malic acid alone, and 100-200 mg / L minced garlic.

[0099] [Table 1]

[0100]

[0101] Example 2

[0102] Evaluation of cashew nut shell liquid (CNSL) and various organic acids using a methane production measurement system using rumen culture fluid

[0103] (1) Sample

[0104] Cashew nut shell liquid (CNSL) extracted from cashew nut shells by low temperature pressing was obtained. Rumen fluid (four-layer gauze filtrate) collected orally from Holstein lactating cows more than 90 days after parturition was used as the culture inoculum. 4 mL of the rumen fluid and 6 mL of artificial saliva (McDougal's artificial saliva) were added for culture.

[0105] (2) Cultivation

[0106] The concentration of CNSL in the test culture medium was set to 100-500 mg / L. CNSL was dissolved in ethanol and added to a Hungate tube to an arbitrary concentration in the liquid. Similarly, pyruvic acid, citric acid, and succinic acid were dissolved in ethanol and added to an arbitrary concentration in the liquid. In order to make the effect of ethanol equal, only ethanol was added to the non-added area (control area). After that, it was left for several hours to evaporate the ethanol, and 50 mg of corn starch, 75 mg of strong flavored feed powder, and 75 mg of timothy hay powder were added as a culture matrix. A total of 10 ml of diluted rumen fluid and artificial saliva were added to these test tubes in the above-mentioned mixing ratio, and nitrogen was blown into the headspace while covering them with butyl rubber caps and plastic screw caps, and anaerobic culture was carried out in an incubator (39°C, 17 hours). Each was set up as a triplicate culture.

[0107] (3) Analysis

[0108] The concentrations of methane and acetic acid in volatile fatty acids were analyzed using FID gas chromatography.

[0109] (4) Results

[0110] (i) Gas production and acetic acid production

[0111] Table 2 shows the results of the gas generation amount and the acetic acid generation amount.

[0112] As in Example 1, reductions in methane production and acetic acid production were observed depending on the CNSL concentration. When 100 mg / L CNSL and 500 to 1000 mg / L citric acid were used in combination, methane production was significantly reduced compared to the addition of 100 mg / L CNSL alone, while no reduction in acetic acid production was observed. No significant increase in methane reduction was observed under the conditions of the combined use of CNSL and pyruvic acid or succinic acid or the addition of each organic acid at the concentration alone.

[0113] [Table 2]

[0114]

[0115] Example 3

[0116] Evaluation of cashew nut shell liquid (CNSL), DL-malic acid, and citric acid using a methane production measurement system using rumen culture fluid

[0117] (1) Sample

[0118] Cashew nut shell liquid (CNSL) extracted from cashew nut shells by low temperature pressing was obtained. Rumen fluid (four-layer gauze filtrate) collected orally from Holstein lactating cows more than 90 days after parturition was used as the culture inoculum. 4 mL of the rumen fluid and 6 mL of artificial saliva (McDougal's artificial saliva) were added for culture.

[0119] (2) Cultivation

[0120] The concentration of CNSL in the test culture medium was set to 100-500 mg / L. CNSL was dissolved in ethanol and added to a Hungate tube to an arbitrary concentration in the liquid. Similarly, DL-malic acid and citric acid were dissolved in ethanol and added to an arbitrary concentration in the liquid. In order to make the effect of ethanol equal, only ethanol was added to the non-added area (control area). After that, it was left for several hours to evaporate the ethanol, and 50 mg of corn starch, 75 mg of strong flavored feed powder, and 75 mg of timothy hay powder were added as the culture matrix. A total of 10 ml of diluted rumen fluid and artificial saliva were added to these test tubes in the above-mentioned mixing ratio, and nitrogen was blown into the headspace while covering them with butyl rubber caps and plastic screw caps, and anaerobic culture was carried out in an incubator (39°C, 17 hours). Each was set up as a triplicate culture.

[0121] (3) Analysis

[0122] The concentrations of methane and acetic acid in volatile fatty acids were analyzed using FID gas chromatography.

[0123] (4) Results

[0124] (i) Gas production and acetic acid production

[0125] Table 3 shows the results of the gas generation amount and the acetic acid generation amount.

[0126] As in Examples 1 and 2, reductions in methane generation and acetic acid generation were observed depending on the CNSL concentration. When 100 mg / L CNSL and 500 to 2000 mg / L DL-malic acid or 500 to 2000 mg / L citric acid were used in combination, methane generation was significantly reduced compared to the addition of 100 mg / L CNSL alone, while no reduction in acetic acid generation was observed. When 500 mg / L citric acid and 500 mg / L DL-malic acid were mixed, a trend was observed in which the methane reduction effect was improved compared to the addition of each organic acid alone.

[0127] [Table 3]

[0128]

Claims

1. A methane production inhibitor for ruminants, characterized in that: The invention contains cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and an organic acid as effective ingredients, and the mass ratio of the amount of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol to the amount of the organic acid is 1:5 to 1:

20.

2. The methane production inhibitor according to claim 1, wherein The organic acid is a carboxylic acid having 3 to 12 carbon atoms.

3. The methane production inhibitor according to claim 1, wherein The organic acid is a hydroxycarboxylic acid having 3 to 10 carbon atoms.

4. The methane production inhibitor according to claim 1, wherein The organic acid is citric acid or malic acid.

5. The methane production inhibitor according to claim 1, wherein Ruminants are cattle. A feed for suppressing methane production for ruminants, comprising the methane production inhibitor according to any one of claims 1 to 5.

7. A method for inhibiting methane production in ruminants, characterized in that: The invention comprises the steps of applying cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol and organic acid to ruminants, wherein the mass ratio of the amount of cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol diphenol applied per ruminant per day to the amount of organic acid applied per ruminant per day is 1:5 to 1:

20.

8. The method according to claim 7, wherein: The organic acid is a carboxylic acid having 3 to 12 carbon atoms.

9. The method according to claim 7, wherein: The organic acid is a hydroxycarboxylic acid having 3 to 10 carbon atoms.

10. The method according to claim 7, wherein: The organic acid is citric acid or malic acid.

11. The method according to claim 7, wherein: Ruminants are cattle.

12. The method according to claim 7, wherein: Cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol are mixed with an organic acid before application.

13. The method according to claim 7, wherein: Cashew nut shell liquid, heat-treated cashew nut shell liquid, anacardic acid, cardanol and / or cardanol and an organic acid are applied separately.

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