CN116669564A

CN116669564A - Composition for reducing methane emissions, method for improving the metabolic efficiency of ruminants and method for administration of methanogenesis inhibitors

Info

Publication number: CN116669564A
Application number: CN202180090172.0A
Authority: CN
Inventors: P·F·芬尼西; D·W·尼夫
Original assignee: Motel Innovation Co ltd
Current assignee: Motel Innovation Co ltd
Priority date: 2020-12-21
Filing date: 2021-12-21
Publication date: 2023-08-29

Abstract

Compositions for reducing methane emissions and/or inhibiting one or more methanogens are provided. The composition comprises an organohalogen compound and an organosulfur compound, preferably bromoform and allicin. The composition may further comprise a polyphenol compound. Animal feeds comprising the composition are also described.

Description

Composition for reducing methane emissions, method for improving the metabolic efficiency of ruminants and method for administration of methanogenesis inhibitors

Technical Field

The present invention relates to compositions for reducing methane emissions and/or for inhibiting one or more methanogenic organisms (i.e., methanogens). The invention also relates to methods for improving the metabolic efficiency of ruminants and methods for administering methanogenesis inhibitors to ruminants, particularly agricultural ruminants.

Background

Ruminants consume feeds containing carbon and nitrogen, which are converted to a variety of substances, including carbon-rich lipids, fats, fatty acids and carbohydrates, as well as nitrogen-rich proteins, nucleic acids, amino acids and nucleotides.

The feeds converted to these substances assist in animal growth and are the major component of a variety of valuable animal products.

Ruminants, such as cattle, sheep and goats, however, are inefficient users of carbon and nitrogen. Methane production from carbon ruminants represents an energy loss, accounting for 2% to 12% of the total energy intake from the feed.

Additionally, ruminants may excrete more than 50% of their nitrogen from the feed, primarily in the form of urinary nitrogen and fecal nitrogen.

Carbon and nitrogen losses from ruminants are both valuable animal growth losses and have significant environmental impact, including nitrogen leaching that allows the strong greenhouse gases methane and nitrous oxide to enter the atmosphere and soil. Feed supplements administered to ruminants to improve animal growth and reduce carbon and nitrogen emissions into the environment are expensive and difficult to administer in an optimal and economical manner.

One of the known methods for reducing methane production in ruminants is by using seaweed. However, feeding animals with large amounts of seaweed may have potential risk factors. One of the risk factors is a decrease in feed intake and performance. Feeding seaweed at higher levels in the diet has resulted in reduced intake of dry matter in beef cattle (Roque et al, 2021) and dairy cows (Roque et al, 2019, stefenoni et al, 2021, muizer et al, 2021). It has been observed that cows often reject or choose to be resistant to seaweed when mixed with their fresh feed, indicating poor palatability of seaweed (muizelar et al 2021). Lower feed intake can also result in lower performance when cows are fed high dose levels of seaweed, as shown by reduced milk production (Roque et al 2019, stefenoni et al 2021, muizer et al 2021). Seaweed is also known to contain high levels of iodine (Makkar et al 2016) and its transfer to livestock products has been studied. Algae (Taxus bacon (Asparagopsis taxiformis)) were fed to beef cattle in diets at content levels of 0.25% and 0.5% resulting in iodine intake of 106 to 225 mg/day per day (Roque et al 2021). This exceeded the recommended daily iodine intake level of about 5 mg/day based on 0.5mg/kg DMI (NRC, 2006) and 10kg DM intake in this study. Iodine transfer into milk is a greater problem. According to Lean et al (2021), feeding Taxus chinensis (Asparagopsis taxiformis) at 0.5% in the diet increased the iodine level 5-fold to 3mg/L.

There is clearly a need to develop improved compositions for reducing methane emissions, in particular wherein the above-mentioned risk factors are reduced and/or wherein the compositions have improved palatability compared to seaweed.

There is also a clear need to improve the metabolic efficiency of ruminants to reduce methane emissions to the environment and nitrogen excretion to the environment, and to improve animal growth levels and increase the amount of valuable animal products.

The present invention addresses this need by providing a feed supplement that improves the metabolic efficiency of ruminants. The feed supplement reduces methane emissions into the environment and nitrogen excretion into the environment and converts energy that would otherwise be converted to emitted methane and otherwise excreted nitrogen into valuable animal products and thus improves the metabolic efficiency of ruminants.

There is clearly a need to improve the administration of feed supplements to ruminants to improve animal growth and reduce carbon and nitrogen losses to the environment.

The present invention addresses this need by providing a composition and methanogenesis inhibitor feed supplement administration that optimally and economically reduces methane emissions and nitrogen excretion and converts energy that would otherwise be converted to emitted methane and otherwise excreted nitrogen into valuable animal products.

Statement of invention

According to the present invention, in a first aspect, there is provided a composition for reducing methane emissions comprising an organohalogen compound and an organosulfur compound.

In some embodiments, the organohalogen compound is an organobromine compound. In a preferred embodiment, the organohalogen compound is bromoform (CHBr ₃ )。

In some embodiments, the organosulfur compound is from a plant of the Allium (Allium) species. In some embodiments, the organosulfur compound is selected from allicin (C ₆ H ₁₀ S ₂ O); diallyl sulfide (C) ₆ H ₁₀ S) S; diallyl disulfide (C) ₆ H ₁₀ S ₂ ) The method comprises the steps of carrying out a first treatment on the surface of the And allyl mercaptan (C) ₃ H ₆ S). In a preferred embodiment, the organosulfur compound is allicin.

In some embodiments, the ratio of organohalogen compound to organosulfur compound is from 1:10 to 1:3500, more preferably from 1:1000 to 1:2500.

In some embodiments, the composition further comprises a polyphenol compound. In some embodiments, the polyphenolic compound comprises or is a bioflavonoid. In some embodiments, the polyphenolic compound comprises naringin, neohesperidin, or a combination thereof.

In some embodiments, the composition is for inhibiting one or more methanogens. In some embodiments, the composition is used to improve the metabolic efficiency of an animal, more particularly a ruminant, such as a cow, goat or sheep.

The inventors have found that the compositions of the present invention exhibit high% inhibition of methanogens when an organohalogen compound (e.g. bromoform) is combined with an organosulfur compound (e.g. allicin).

In particular, it has been shown that organohalogen compounds and organosulfur compounds can act synergistically to reduce methane production. Synergistic combinations provide effects greater than the sum of the individual components. The inventors have additionally found that compositions comprising an organohalogen compound (e.g. bromoform) and a powder mixture comprising an organosulfur compound and a polyphenol compound (e.g. NXRH214 powder) also lead to effective inhibition of methanogens.

Also according to the invention, in a second aspect, is an animal feed comprising the composition of the invention or as otherwise disclosed herein.

Also according to the invention, in a third aspect, is the use of the composition of the invention or the animal feed of the invention for reducing methane emissions and/or for inhibiting methanogens and/or improving the metabolic efficiency of an animal.

Also according to the invention in a fourth aspect is a method of reducing methane emissions, the method comprising administering to an animal, more particularly a ruminant, a composition or animal feed of the invention. In some examples, the ruminant is a cow, goat or sheep.

Also according to the invention, in a fifth aspect, is a method of inhibiting one or more methanogens, the method comprising administering to an animal, more particularly a ruminant, a composition or animal feed of the invention. In some examples, the ruminant is a cow, goat or sheep.

Also according to the invention, in a sixth aspect, is a method of improving the metabolic efficiency of an animal, the method comprising administering to the animal, more particularly a ruminant, a composition or animal feed of the invention. In some examples, the ruminant is a cow, goat or sheep.

Also disclosed herein are compositions for inhibiting one or more methanogens comprising an organohalogen compound and an organosulfur compound. The organohalogen compound and the organosulfur compound are as further described herein.

Also disclosed herein are compositions for improving metabolic efficiency in an animal, the compositions comprising an organohalogen compound and an organosulfur compound. The organohalogen compound and the organosulfur compound are as further described herein.

Also disclosed herein are methods of reducing nitrogen excretion by a ruminant and/or reducing methane emissions by a ruminant and/or increasing nitrogen-rich and carbon-rich materials in a ruminant, the method comprising the step of administering to the ruminant an effective amount of at least one type of methanogenesis inhibitor. In one embodiment, the method comprises administering any of the compositions disclosed herein to the ruminant.

In one embodiment, the methanogenesis inhibitor is selected from the group consisting of: an organohalogen compound; marine macroalgae enriched in organic halogen; an organic sulfur compound; plants rich in organic sulfur; a polyphenol compound; and polyphenol-enriched plants.

In one embodiment, the organohalogen compound is selected from: CH (CH) ₃ Cl；CH ₃ Br；CH ₃ I；CH ₂ Cl ₂ ；CH ₂ Br ₂ ；CH ₂ I ₂ ；CHCl ₃ ；CHBr ₃ ；CHI ₃ ；CCl ₄ ；CBr ₄ ；CH ₂ ClBr；CH ₂ ClI；CH ₂ BrI；CHBr ₂ Cl；CHBrI ₂ ；CHBrClI；CHBr ₂ I；CHBrCl ₂ ；CH ₃ CH ₂ Br；CH ₃ CH ₂ I；CH ₃ CH ₂ CH ₂ I；CH ₃ (CH ₂ ) ₃ I；CH ₃ (CH ₂ ) ₄ Br；CH ₃ (CH ₂ ) ₄ I；(CH3) ₂ CHI；CH ₃ CH ₂ CH(CH ₃ )I；(CH ₃ ) ₂ CHCH ₂ I；BrCH ₂ CH ₂ Br；ClCH＝CCl ₂ The method comprises the steps of carrying out a first treatment on the surface of the And CH (CH) ₃ CH ₂ CH ₂ CH ₂ I。

In one embodiment, the organohalogen-rich marine macroalgae is selected from the group consisting of: radix seu radix Ophiopogonii (Asparagopsis armata); taxus-like sea-door winter (Asparagopsis taxiformis); a Dictyota (Dictyota) species; a sphingosine (oendonium) species; ulva (Ulva) species; and cladophora (Cladophora patentiramea).

In one embodiment, the organosulfur compound is selected from: organic sulfur secondary metabolites; allicin (C) ₆ H ₁₀ S ₂ O); diallyl sulfide (C) ₆ H ₁₀ S) S; diallyl disulfide (C) ₆ H ₁₀ S ₂ ) The method comprises the steps of carrying out a first treatment on the surface of the And allyl mercaptan (C) ₃ H ₆ S)。

In one embodiment, the organosulfur-rich plant is selected from the Allium species: garlic (Allium sativum); green Chinese onion (Allium ampeloprasum); and onion (Allium cepa).

In one embodiment, the polyphenolic compound is selected from the group consisting of: flavonoids; bioflavonoids; non-bioflavonoids. The at least one polyphenolic compound may, for example, comprise at least one bioflavonoid; -a flower flavin; flavone; flavonols; flavanones; a flavanonol; a flavan; anthocyanin; isoflavane; new flavanthamine; isoflavone; procyanidins; phenolic acid; hydroxycinnamic acid; coumarin; stilbenes; anthraquinone; lignans; lignin; tannin; a polyphenol protein; catechin; rutin; robinia pseudoacacia; genistein; kaempferol; gallocatechin; catechin gallate; epicatechin; epigallocatechin; epicatechin gallate; quercetin; epicatechin (allocalin); gallocatechin gallate; epicatechin; epigallocatechin; epicatechin gallate; epigallocatechin gallate; kaempferol; quercetin; naringin; neohesperidin; eriocitrin; naringin; naringenin; hesperidin; rhus verniciflua fordii glycoside (roifolin); myrosin; melissa glucoside; hesperetin; poncirin; epicatechin; gallocatechin; epigallocatechin; coumaric acid; cinnamic acid; gallic acid; ellagic acid; protocatechuic acid; chlorogenic acid; caffeic acid; ferulic acid; punicalagin; pomegranate rind tannagin.

In one embodiment, the polyphenol-enriched plant is selected from the group consisting of: allium species; a Brassica (Brassica) species; a camellia (Camelia) species; a Capsicum (Capsicum) species; citrus (Citrus) species; citrus aurantium (Citrus aurantium); cucumber (Cucumis) species; malus (Malus) species; a Musa (Musa) species; a Phaseolus (Phaseolus) species; a prune (prune) species; a Punica (Punica) species; pear (Pyrus) species; solanum (Solanum) species; and Vaccinium species.

In another aspect, the present invention provides a method for reducing nitrogen and carbon emissions excreted by a ruminant and increasing valuable nitrogen-and carbon-rich materials in a ruminant, the method comprising the step of administering to the animal a feed supplement as described herein or a feed as described herein.

Also disclosed herein are methods of reducing nitrogen excretion by a ruminant and/or reducing methane emissions by a ruminant and/or increasing nitrogen-and carbon-rich materials in a ruminant, the methods comprising progressively administering to the ruminant an effective amount of at least one type of methanogenesis inhibitor. In one embodiment, the method comprises administering to the ruminant a composition as disclosed herein.

In one embodiment, the methanogenesis inhibitor is administered stepwise with at least one dose, the percentage of which on the ruminant weight is selected from the group consisting of: 0.1%;0.2%;0.3%;0.4%;0.5%;0.6%;0.7%;0.8%;0.9%;1.0%;1.1%;1.2%;1.3%;1.4%;1.5%;2.0%;2.5%;3.0%;3.5%;4.0%;4.5%;5.0%; and 10%.

In one embodiment, the step-wise administration of the methanogenesis inhibitor is with at least one dose, the percentage of which in the feed weight of the ruminant is selected from: 0.1%;0.2%;0.3%;0.4%;0.5%;0.6%;0.7%;0.8%;0.9%;1.0%;1.1%;1.2%;1.3%;1.4%;1.5%;2.0%;2.5%;3.0%;3.5%;4.0%;4.5%;5.0%; and 10%.

In one embodiment, the stepwise administration has at least one interval between successive doses, the interval selected from the group consisting of: 1 minute; 1 hour; 1 day; for 2 days; 3 days; 4 days; for 5 days; for 6 days; 7 days; for 10 days; 2 weeks; 3 weeks; 4 weeks; for 6 weeks; 2 months; 3 months; 4 months; 6 months; 9 months; and 12 months.

Brief Description of Drawings

Figures 1 and 2 show the% methane inhibition of various compositions of the invention after incubation with a culture of methanococcus maritimus (Methanococcus maripaludis).

Detailed Description

According to the present invention there is provided a composition for reducing methane emissions comprising an organohalogen compound and an organosulfur compound. The compositions may also be used to inhibit one or more methanogens. Any "composition" referred to herein may also be referred to as an "animal feed supplement".

In some embodiments, the one or more methanogens have methanobacteria (methanobacteria), methanosarcina (Methanosarcina), methanobrevibacteria (methanobrevulbater), methanosarcina (Methanosarcina), methanobagrus (methanocullus), methanosphaera (Methanosphaera), methanogranella (methanonocorkulum), methanovesiculus (Methanofollis), methanogenelium (methanogenelium), methanomicrobacteria (methanomicrobacteria), methanofire (Methanopyrus), methanobreguella, methanomans (methanomergua), methanothermophilus (methanothermophilic bacteria), or Methanococcus (methanonococcus). In some embodiments, the one or more methanogens are selected from the group consisting of methanobacterium formate (Methanobacterium formicicum), methanobacterium buchneri (Methanobacterium bryantii), methanobacterium rumen (Methanobrevibacter ruminantium), methanobacterium migrainae (Methanobrevibacter millerae), methanobacterium australis (Methanobrevibacter olleyae), methanobacterium moti (Methanomicrobium mobile), methanobacterium australis (Methanoculleus olentangyi), methanobacterium sarcina bararyophyllum (Methanosarcina barkeri), methanobacterium borgpi (Methanobrevibacter boviskoreani), methanobacterium beijing (Methanobacterium beijingense), methanobacterium jersey (Methanoculleus marisnigri), methanobacterium breveticus (Methanoculleus bourgensis), methanosarcina mahogany (Methanosarcina mazei), methanobacterium gossypii (Methanobrevibacter gottschalkii), methanobacterium (Methanobrevibacter thaueri), methanobacterium smithii (Methanobrevibacter smithii), methanobacterium stevens (Methanosphaera stadtmanae), methanobacterium Wo Sishi (Methanobrevibacter woesei), methanobacterium wovens (Methanobrevibacter wolinii). In some examples, the one or more methanogens is methanococcus maritimus (Methanococcus maripaludis).

Hereinafter, the present invention will be described according to preferred embodiments thereof with reference to the accompanying description. It should be understood, however, that the description is limited to the preferred embodiments of the invention only for the purpose of facilitating the discussion of the invention, and that various modifications may be devised by those skilled in the art without departing from the scope of the appended claims.

Organic halogen compounds

The compositions of the present invention comprise an organohalogen compound (i.e., at least one organohalogen compound).

The organic halogen compound is a halogen-containing organic compound. In some embodiments, the organohalogen compound is C ₁ -C ₆ An alkyl halide compound. In some embodiments, the organohalogen compound comprises chlorine, bromine, iodine, or a combination thereof. In some embodiments, the organohalogen compound comprises CH ₃ Cl；CH ₃ Br；CH ₃ I；CH ₂ Cl ₂ ；CH ₂ Br ₂ ；CH ₂ I ₂ ；CHCl ₃ ；CHBr ₃ ；CHI ₃ ；CCl ₄ ；CBr ₄ ；CH ₂ ClBr；CH ₂ ClI；CH ₂ BrI；CHBr ₂ Cl；CHBrI ₂ ；CHBrClI；CHBr ₂ I；CHBrCl ₂ ；CH ₃ CH ₂ Br；CH ₃ CH ₂ I；CH ₃ CH ₂ CH ₂ I；CH ₃ (CH ₂ ) ₃ I；CH ₃ (CH ₂ ) ₄ Br；CH ₃ (CH ₂ ) ₄ I；(CH3) ₂ CHI；CH ₃ CH ₂ CH(CH ₃ )I；(CH ₃ ) ₂ CHCH ₂ I；BrCH ₂ CH ₂ Br；ClCH＝CCl ₂ The method comprises the steps of carrying out a first treatment on the surface of the And CH (CH) ₃ CH ₂ CH ₂ CH ₂ I. In some embodiments, the organohalogen compound is trihalomethane. In some embodiments, the organohalogen compound is an organobromine compound, more preferably wherein the organohalogen compound is bromoform (CHBr ₃ )。

The biological source of the organohalogen compound is an organohalogen-rich marine macroalgae. For example, the organohalogen-rich marine macroalgae comprises at least one marine macroalgae species selected from the group consisting of: radix seu radix Ophiopogonii (Asparagopsis armata); taxus-like sea-door winter (Asparagopsis taxiformis); a Dictyota (Dictyota) species; a sphingosine (oendonium) species; ulva (Ulva) species; and cladophora (Cladophora patentiramea). Thus, in some embodiments, the organohalogen compound is derived from an organohalogen-rich marine microalgae, for example selected from: radix seu radix Ophiopogonii (Asparagopsis armata); taxus-like sea-door winter (Asparagopsis taxiformis); a Dictyota (Dictyota) species; a sphingosine (oendonium) species; ulva (Ulva) species; and cladophora (Cladophora patentiramea).

In other embodiments, the organohalogen compound is produced by bacteria, fungi, and cyanobacteria. For example, the bacteria include one selected from the group consisting of: streptomyces sp and Flavobacterium galactovorans Zhuo Beier (Zobellia galactanivorans). For example, the fungus includes one fungus selected from the group consisting of: pyricularia oryzae (Pyricularia oryzae), curvularia curvularia (Curvularia inaequalis), pyrenophora pumila (Pyrenophora tritici-repentis) and Embellisia didymospora. For example, cyanobacteria include one selected from the group consisting of: red sea Shu Maozao (Trichodesmium erythraeum), synechococcus sp and deep sea single cell blue algae (Acaryochloris marina).

In other embodiments, the organohalogen is synthetic, i.e., the organohalogen is chemically synthesized. In other embodiments, the organohalogen is produced by recombinant yeast.

In some embodiments, the concentration of the organohalogen compound in the composition is greater than 100nM, or greater than 110nM, or greater than 120nM, or greater than 130nM, or greater than 140nM, or greater than 150nM. In some embodiments, the concentration of the organohalogen compound in the composition is less than 10000nM or less than 1000nM or less than 500nM or less than 200nM, or less than 175nM, or less than 160nM, or less than 150nM, or less than 140nM, or less than 130nM. In some embodiments, the concentration of the organohalogen compound in the composition is 100nM to 10000nM,100nM to 1000nM,100nM to 500nM,100nM to 300nM,100nM to 200nM, or 110nM to 175nM. In some examples, the organohalogen compound is an organobromine compound, preferably bromoform.

Organic sulfur compounds

The compositions of the present invention comprise an organic sulfur compound (i.e., at least one organic sulfur compound).

The organic sulfur compound is an organic compound containing sulfur. In certain embodiments, each organosulfur compound may be independently selected from the group consisting of a thioether, thioester, thioacetal, thiol, disulfide, polysulfide, sulfoxide, sulfone, thiosulfinate, sulfonimide, sulfimide, sulfone diimine, thioketone, thioaldehyde, sulfonium, sulfone alkene, thiocarboxylic acid (including dithiocarboxylic acids), sulfonic acid, sulfinic acid, sulfenanic acid, sulfonate, sulfinate, sulfenate, sulfonamide, sulfinamide, sulfenamide, sulfonium compound, oxosulfonium compound, sulfonium salt, oxosulfonium salt, thiocarbonyl endoium salt, sulfane (sulfane), and persulfate (persulfurane). In certain embodiments, each organosulfur compound is independently selected from thioesters, sulfoxides, sulfides, disulfides, polysulfides (including trisulfides), and thiols. In certain embodiments, each organosulfur compound is independently selected from thioesters, sulfoxides, sulfides, disulfides, and polysulfides (including trisulfides). In certain embodiments, each organosulfur compound is independently selected from disulfides and polysulfides (including trisulfides). In certain embodiments, each organosulfur compound is a disulfide.

In some embodiments, the organosulfur compound is from a plant of the Allium (Allium) species. In some embodiments, the organosulfur compound is a disulfide compound, and more specifically a diallyl disulfide compound.

In certain embodiments, each organosulfur compound is independently selected from allicin, allyl disulfide, diallyl trisulfide, s-allyl cysteine, vinyl dithiines (3-vinyl-4H-1, 2-dithiines) and 2-vinyl-4H-1, 3-dithiines (2-vinyl-4H-1, 3-dithiines), and diallyl disulfide.

In some embodiments, the organosulfur compound is selected from allicin (C ₆ H ₁₀ S ₂ O); diallyl sulfide (C) ₆ H ₁₀ S) S; diallyl disulfide (C) ₆ H ₁₀ S ₂ ) The method comprises the steps of carrying out a first treatment on the surface of the And allyl mercaptan (C) ₃ H ₆ S). In certain embodiments, the at least one organosulfur compound is or includes allicin.

Allicin is of formula C ₆ H ₁₀ OS ₂ And an organic sulfur compound having a structure shown below.

Organosulfur compounds, such as allicin, may be obtained, for example, from garlic or another Allium species. For example, the organosulfur compound (e.g., allicin) may be obtained from an extract of Allium species such as Allium sativum (Allium sativum). The term extract includes aqueous extracts, non-aqueous extracts, alcoholic extracts, concentrates, oils, infusions, powders, granules and combinations of two or more thereof. For example, the organic sulfur compound (e.g., allicin) may be obtained from raw garlic, dried garlic, or a combination thereof. The organosulfur compounds (e.g., allicin) may be derived, for example, from any Allium subspecies and varieties currently known or later discovered, such as garlic (Allium sativum), allium ursinum (Allium ursinum), allium fistulosum (Allium fistulosum), allium cepa (Allium cepa), and Mao Cong (Allium tricoccum). For example, the organosulfur compounds (e.g., allicin) can be independently derived from garlic of the sub-species ophiocorodon (hard necked garlic) and garlic of the sub-species sativum (soft necked garlic). For example, organosulfur compounds (such as allicin) may be independently derived from porcelain garlic (garlic), cucurbit garlic (rocambole garlic), purple garlic (purple stripe garlic), marble Dan Ziwen garlic (marbled purple stripe garlic), glazed purple garlic (glazed purple stripe garlic), artichoke garlic (aromachanic garlic), silver skin garlic (silverskin garlic), asiatic garlic (asiatic garlic), cape garlic (turban garlic), and crioll garlic (creoll garlic). In particular, organosulfur compounds (such as allicin) are available from garlic (Allium sativum).

For example, allium plants (Allium) that can be derived from organosulfur compounds (e.g., allicin) can have been treated or processed. For example, allium plants (Allium) can be "aged" or "black" (e.g., aged or black garlic) obtained by storing Allium plants (Allium) under controlled conditions and heating at specific temperatures, humidity and solvents, e.g., over several days or weeks, after undergoing Maillard or browning reactions, darkening the garlic cloves. For example, allium plants (Allium) can be "dried" or "dehydrated" by heating fresh or unaged garlic to a temperature of 30 ℃ to 120 ℃ and to a moisture content of about 3% to 10% with or without converting or converting its ingredients to a different compound. For example, allium plants (Allium) may be "fresh" or "unaged" (e.g., fresh or unaged garlic) that have not been intentionally subjected to special treatments or processes to convert or convert their components to different compounds. Fresh or unaged Allium plants (Allium) may, for example, have been treated or processed to remove odors (deodorized) (such as deodorized garlic extracts). Generally, encapsulation or coating methods can be employed to mask or reduce odors. Alternatively or additionally, taste masking ingredients such as green tea, parsley, basil, spinach, and the like may be added to mask or reduce odors in the composition.

The organosulfur compound (e.g., allicin) may or may not be isolated and/or purified prior to incorporation into the compositions described herein. Thus, in certain embodiments, the compositions described herein may comprise raw garlic, dry garlic, and/or garlic extract. In other embodiments, the organosulfur compound (e.g., allicin) is chemically synthesized. In certain embodiments, the allicin may be obtained by treating a natural source of alliinase to release alliinase, contacting the treated source of alliinase with alliin, thereby enzymatically converting alliin to allicin, and optionally extracting allicin. Suitable processes are further described, for example, in WO03/004668, the contents of which WO03/004668 are incorporated herein by reference.

In other embodiments, the organosulfur compound, such as allicin, may be synthetic, i.e., chemically synthesized.

In some embodiments, the concentration of the organosulfur compound in the composition is greater than 10 μm, or greater than 100 μm, or greater than 150 μm, or greater than 175 μm, or greater than 200 μm, or greater than 225 μm, or greater than 250 μm, or greater than 275 μm. In some embodiments, the concentration of the organosulfur compound in the composition is less than 350 μm, or less than 325 μm, or less than 300 μm, or less than 275 μm, or less than 250 μm, or less than 225 μm. In some embodiments, the concentration of the organosulfur compound in the composition is from 10 μm to 350 μm, from 100 μm to 350 μm, from 150 μm to 350 μm, and in some examples, from 200 to 300 μm.

Ratio of organohalogen compound to organosulfur compound

In some embodiments, the ratio of organohalogen compound to organosulfur compound in the composition is preferably from 1:10 to 1:3500. In some embodiments, the ratio of organohalogen compound to organosulfur compound in the composition is from 1:50 to 1:3500, or from 1:100 to 1:3500, or from 1:500 to 1:3500, or from 1:750 to 1:3500, or from 1:1000 to 1:3500, or from 1:1500 to 1:3500, or from 1:2000 to 1:3500. In some embodiments, the ratio of organohalogen compound to organosulfur compound in the composition is from 1:500 to 1:3000, or from 1:500 to 1:2750, or from 1:500 to 1:2500, or from 1:500 to 1:2000, or from 1:500 to 1:1500. In preferred embodiments, the ratio of organohalogen compound to organosulfur compound in the composition is from 1:750 to 1:3000, or from 1:1000 to 1:2500.

In some examples, the organohalogen is an organobromine compound, such as bromoform, and the organosulfur compound is a disulfide, such as allicin.

-polyphenol compounds

The compositions of the present invention may further comprise a polyphenolic compound (i.e., one or more polyphenolic compounds).

The term phenol refers to a chemical compound comprising a hydroxyl group (-OH) directly bonded to an aromatic hydrocarbon group. The term polyphenol compound refers to a compound comprising more than one phenolic group. The polyphenolic compounds described herein may include bioflavonoids, non-bioflavonoid polyphenolic compounds, or combinations thereof. The at least one polyphenolic compound may, for example, comprise at least one bioflavonoid.

The term biological yellowKetone refers to a class of plant and fungal secondary metabolites and has a general structure of 15 carbon skeleton consisting of two benzene rings (A and B) and a heterocycle (C), sometimes abbreviated as C ₆ -C ₃ -C ₆ . Bioflavonoids are thus polyphenols. The term bioflavonoids includes anthocyanins (including flavones and flavonols), flavanones, flavanonols, flavans and anthocyanins. The term bioflavonoids also includes compounds having a flavone backbone (2-phenyl-1, 4-benzopyrone), an isoflavane backbone (3-phenylchromen-4-one) or a neoflavane backbone (4-phenylcoumarin). The term non-bioflavonoid polyphenolic compound refers to other classes of polyphenolic compounds known in the art which do not fall within the definition of the term bioflavonoid as described herein. The term non-bioflavonoid polyphenol compound includes polyphenol compounds comprising 6 carbons or more, 7 carbons or more, 8 carbons or more, 9 carbons or more, 10 carbons or more, 13 carbons or more, 14 carbons or more, 16 carbons or more, 18 carbons or more, or 30 carbons or more. The term non-bioflavonoid polyphenolic compounds includes, but is not limited to, polyphenolic acids (C ₆ -C ₁ Structure), stilbene compounds (C) ₆ -C ₂ -C ₆ Structure), anthraquinone (C) ₆ -C ₂ -C ₆ Structure) and lignan ((C) ₆ -C ₃ ) ₂ Structure). In some embodiments, the non-bioflavonoid polyphenolic compounds are plant polymers including, but not limited to, lignin, catechol melanin, polyflavans (flavans), polyphenol proteins, and polyphenols. In certain embodiments, the one or more bioflavonoids are each independently selected from the group consisting of anthocyanins (including flavones and flavonols), flavanones (including flavanone glycosides), flavanonols, flavans, isoflavones, anthocyanidins, and procyanidins. In certain embodiments, each of the one or more bioflavonoids is independently selected from the group consisting of an anthoxanthin and a flavanone (including flavanone glycosides). In certain embodiments, all bioflavonoids are anthocyanins and/or flavanones. In certain embodiments, the one or more bioflavonoids are independently flavones or flavones. In certain embodiments, all bioflavonoids are flavones and/or flavanones. Flavone and flavanoneFor example, flavone glycosides and flavanone glycosides, respectively. In certain embodiments, the one or more bioflavonoids is a flavanone. In certain embodiments, all bioflavonoids are flavones. In certain embodiments, the one or more bioflavonoids is a flavanone glycoside. In certain embodiments, all bioflavonoids are flavanone glycosides. The one or more bioflavonoids may be selected, for example, from naringin, neohesperidin, eriocitrin, naringin, hesperidin, rhoifolin, myrosin, melissa glucoside, hesperetin, poncirin, catechin, rutin, locust element, genistein, kaempferol, quercetin, epicatechin, gallocatechin, epigallocatechin, catechin gallate, epicatechin gallate, epigallocatechin gallate and gallocatechin gallate. In certain embodiments, the one or more bioflavonoids include naringin and neohesperidin. In certain embodiments, the one or more bioflavonoids is a combination of naringin and neohesperidin. In certain embodiments, the one or more bioflavonoids comprise one or more of catechin, rutin, locust element, genistein, kaempferol, gallocatechin, catechin gallate, epicatechin, epigallocatechin, epicatechin gallate, and quercetin. In certain embodiments, the one or more bioflavonoids comprise one or more of catechin, rutin, locust bean gum, genistein, and kaempferol. In certain embodiments, the one or more bioflavonoids is a combination of catechin, rutin, locust bean gum, genistein, and kaempferol. In certain embodiments, the one or more bioflavonoids comprise one or more of gallocatechin, catechin gallate, epicatechin, epigallocatechin, epicatechin gallate, gallocatechin gallate, epigallocatechin gallate, kaempferol, and quercetin. In certain embodiments, the one or more bioflavonoids comprise gallocatechin, catechin gallate, epicatechin, epigallocatechin, epicatechin gallate, kaempferol, and quercetin One or more of the following. In certain embodiments, the one or more bioflavonoids is a combination of gallocatechin, catechin gallate, epicatechin, epigallocatechin, epicatechin gallate, gallocatechin gallate, kaempferol, and quercetin. In certain embodiments, the one or more bioflavonoids is a combination of gallocatechin, catechin gallate, epicatechin, epigallocatechin, epicatechin gallate, kaempferol, and quercetin.

In some embodiments, the polyphenols comprise one or more non-bioflavonoid polyphenolic compounds. In some embodiments, the one or more non-bioflavonoid phenolic compounds are each independently selected from the group consisting of phenolic acids, stilbenes, anthraquinones, lignans, tannins, polyphenolic proteins, and polyphenols. In certain embodiments, each of the one or more non-bioflavonoid polyphenolic compounds is independently selected from tannins and polyphenols. In certain embodiments, all of the non-bioflavonoid polyphenolic compounds are tannins and/or polyphenols.

The compositions described herein comprise one or more polyphenolic compounds. For example, the composition may comprise two or more polyphenolic compounds or three or more polyphenolic compounds or four or more polyphenolic compounds or five or more or six or more or seven or more or eight or more or nine or more or ten or more polyphenolic compounds. For example, the composition may comprise one, two, three, four or five polyphenolic compounds. The compositions described herein comprise one or more bioflavonoids. For example, the composition may comprise two or more bioflavonoids, or three or more bioflavonoids, or four or more bioflavonoids, or five or more or six or more or seven or more or eight or more or nine or more or ten or more bioflavonoids. For example, the composition may comprise one, two, three, four or five bioflavonoids. For example, the composition may comprise two bioflavonoids, which may be naringin and neohesperidin. In another example, the composition may comprise five bioflavonoids, which may be catechin, rutin, locust in, genistein and kaempferol. In alternative embodiments, the composition may comprise seven bioflavonoids, which may be gallocatechin, catechin gallate, epicatechin, epigallocatechin, epicatechin gallate, kaempferol, and quercetin. In alternative embodiments, the composition may comprise nine bioflavonoids, which may be gallocatechin, catechin gallate, epicatechin, epigallocatechin, epicatechin gallate, gallocatechin gallate, epigallocatechin gallate, kaempferol, and quercetin.

One or more polyphenolic compounds, such as one or more bioflavonoids, may be obtained, for example, from parts of plants (e.g., fruits or vegetables). For example, flavonols can be obtained from tomatoes, beans, almonds and/or radishes. For example, flavan-3-ols may be obtained from peach, plum, strawberry, and/or green tea. For example, the flavone may be obtained from watermelon and/or capsicum. For example, flavanones may be obtained from fruits of Citrus (Citrus) species. For example, the anthocyanin may be obtained from blueberry, banana, strawberry, cranberry and/or plum. One or more polyphenolic compounds, e.g., one or more bioflavonoids, may be obtained, for example, from Citrus (Citrus) species fruits such as orange, lemon, grapefruit, or lime. In particular, one or more polyphenolic compounds, such as one or more bioflavonoids, may be obtained from orange. One or more polyphenolic compounds, e.g. one or more bioflavonoids, may be obtained, for example, from fruits of Punica (Punica) species, such as Punica granatum (Punica granatum) or suo kot la Punica granatum (Punica protopunica). In particular, one or more polyphenolic compounds, such as one or more bioflavonoids, may be obtained from Punica granatum (Punica granatum).

One or more polyphenolic compounds, such as one or more bioflavonoids, may be obtained, for example, from parts (e.g., leaves) of plants of the genus Camellia (Camellia) species, plants of the species Camellia genus (Camellia) such as tea tree (Camellia sinensis), marble tea (Camellia taliensis), tea-oil Camellia (Camellia oleifera), hong Kong Mao Ruicha (Camellia assimilis), red Camellia (Camellia azalea), short-post tea (Camellia brevistyla), tail She Shancha (Camellia caudata), zhejiang red Camellia (Camelllia chekiangoleosa), golden Camellia (Camellia chrysantha), bao Shejin scented tea (Camellia chrysanthoides), lian She Shancha (Camellia connata), red-skin brown fruit tea (Camellia crapnelliana), lipium tea (Camellia crapnelliana), pulse-exhibiting golden Camellia (Camellia crapnelliana), camellia crapnelliana (Camellia crapnelliana), yellow-flower tea (Camellia flava) callia fleuryi, calomelas stamen tea (Camellia crapnelliana), calophyllum inophyllum tea (Camellia crapnelliana), philippia crudus tea (Camellia crapnelliana), calophyllum inophyllum (Camellia crapnelliana), camellia sinensis (Camellia grijisii), hengchun tea (Camellia crapnelliana), dongshan tea (Camellia crapnelliana), hongkong tea (Camellia crapnelliana), yunnan Burma tea (Camellia crapnelliana), japanese Camellia (Camellia crapnelliana), camellia sinensis (Camellia kisii), camellia sinensis (Camellia crapnelliana), camellia sinensis (Camellia miyagii), camellia sinensis (Camellia crapnelliana), herba capable of being high mountain tea (Camellis nokoensis), fine flower short core tea (Camellia parviflora), southwest Camellia (Camellia pitardii), light red bud Camellia (Camellia pleurocarpa), winding Tian Gonghua Camellia oleifera (Camellia polyodonta), thick broad-leaved Camellia (Camellia pubupetala), yunnan Camellia (Camellia reticulata), camellia rosiflora, snow mountain tea (Camellia rusticana), willow Camellia (Camellia salicifolia), angjiang Camellia (Camellia saluenensis), camellia sinensis (Camellia sasanqua), safflower Camellia (Camellia semiserrata), general energy herba high mountain tea (Camellis trasnokoensis), yunnan even core tea (Camellia tsaii), dongxing Camellia chrysantha (Camellia tunghinensis), vietnam Camellia (Camellia vietnamensis), williams hybrid Camellia (Camellia x williamsii) and monkey-wood Camellia (Camellia yunnanensis). In particular, one or more bioflavonoids may be obtained from tea tree (Camellis sinensis) (tea plants). Any subspecies or variety of tea tree (Camellia sinensis) may be used. Portions (e.g., leaves) of tea tree (Camellia sinensis) may be untreated or may be treated, for example, by steaming, withering, rolling, oxidizing, fermenting, and/or drying. One or more polyphenolic compounds, such as one or more bioflavonoids, may be obtained, for example, from green tea (tea tree (Camellia sinensis)) leaves.

For example, one or more polyphenolic compounds, such as one or more bioflavonoids, may be obtained from an extract of a Citrus (Citrus) species fruit, a Punica (Punica) species fruit, or a portion of a Camellia (Camellia) species plant. The term extract includes aqueous extracts, non-aqueous extracts, alcoholic extracts, concentrates, oils, infusions, powders, granules and combinations of two or more thereof. For example, one or more polyphenolic compounds, such as one or more bioflavonoids, may be obtained from dried Citrus (Citrus) fruits, dried Punica (Punica) fruits, or dried Camellia (Camellia) plant parts (e.g., leaves). For example, one or more polyphenolic compounds, such as one or more bioflavonoids, may be obtained from raw Citrus (Citrus) fruits, raw Punica (Punica) fruits, or raw Camellia (Camellia) plant parts (e.g., leaves).

One or more polyphenolic compounds, such as one or more bioflavonoids, may or may not be isolated and/or purified prior to incorporation into the compositions described herein. Thus, in certain embodiments, the compositions described herein can comprise raw Citrus (Citrus) fruit, dried Citrus (Citrus) fruit, and/or Citrus (Citrus) fruit extract, or raw pomegranate (Punica) fruit, dried pomegranate (Punica) fruit extract, or raw Camellia (Camellia) plant, dried Camellia (Camellia) plant, and/or Camellia (Camellia) plant extract.

In other embodiments, one or more polyphenolic compounds, such as one or more bioflavonoids, may each be independently chemically synthesized.

In certain embodiments, the compositions described herein comprise two polyphenolic compounds, e.g., two bioflavonoids. The ratio of the first polyphenol compound to the second polyphenol compound, e.g., the first bioflavonoid to the second bioflavonoid, may be, for example, in the range of about 0.5:5 to about 3:1. For example, the ratio of the first polyphenol compound to the second polyphenol compound, e.g., the first bioflavonoid to the second bioflavonoid, may be in the range of about 0.5:5 to about 2.5:1, or about 0.5:5 to about 2:1, or about 0.5:5 to about 1.5:1, or about 0.5:5 to about 1:1. For example, the ratio of the first polyphenol compound to the second polyphenol compound, e.g., the first bioflavonoid to the second bioflavonoid, may be in the range of about 1:5 to about 3:1, or about 1.5:5 to about 3:1, or about 2:5 to about 3:1, or about 2.5:5 to about 3:1, or about 3:5 to about 3:1, or about 3.5:5 to about 3:1, or about 4:5 to about 3:1, or about 4.5:5 to about 3:1, or about 5:5 to about 3:1. The ratio is preferably 2:1.

In certain embodiments, the compositions described herein comprise naringin and neohesperidin. In certain embodiments, the at least one polyphenol comprises a major portion of naringin, neohesperidin, or a combination thereof, wherein the major portion refers to at least 50 wt%, or at least 60 wt%, or at least 70 wt%, or at least 80 wt%, or at least 90 wt% of the total weight of the polyphenol compounds. The ratio of naringin to neohesperidin can be, for example, in the range of about 0.5:5 to about 3:1. For example, the ratio of naringin to neohesperidin can range from about 0.5:5 to about 2.5:1, or from about 0.5:5 to about 2:1, or from about 0.5:5 to about 1.5:1, or from about 0.5:5 to 1:1. For example, the ratio of naringin to neohesperidin can range from about 1:5 to about 3:1, or from about 1.5:5 to about 3:1, or from about 2:5 to about 3:1, or from about 2.5:5 to about 3:1, or from about 3:5 to about 3:1, or from about 3.5:5 to about 3:1, or from about 4:5 to about 3:1, or from about 4.5:5 to about 3:1, or from about 5:5 to about 3:1. The ratio is preferably 2:1.

In certain embodiments, the ratio of total organic sulfur compounds to total polyphenolic compounds (e.g., the ratio of total organic sulfur compounds to total bioflavonoids) can range from about 16:1 to about 1:30. For example, the ratio of total organosulfur compounds to total polyphenolic compounds (e.g., the ratio of total organosulfur compounds to total bioflavonoids) can range from about 15:1 to about 1:30, or from about 14:1 to about 1:30, or from about 13:1 to about 1:30, or from about 12:1 to about 1:30, or from about 10:1 to about 1:30, or from about 16:1 to about 1:16. For example, the ratio of total organosulfur compounds to total polyphenolic compounds (e.g., the ratio of total organosulfur compounds to total bioflavonoids) can range from about 9:1 to about 1:25, or from about 8:1 to about 1:20, or from about 7:1 to about 1:15, or from about 6:1 to about 1:10, or from about 5:1 to about 1:8, or from about 4:1 to about 1:7, or from about 3:1 to about 1:6, or from about 2:1 to about 1:5, or from about 1:1 to about 1:4. For example, the ratio of total organic sulfur compounds to total polyphenolic compounds (e.g., the ratio of total organic sulfur compounds to total bioflavonoids) can be in the range of about 1:1 to about 1:3, or about 2:1 to about 1:4. For example, the ratio of total organic sulfur compounds to total polyphenolic compounds (e.g., the ratio of total organic sulfur compounds to total bioflavonoids) may be about 1:3.

In certain embodiments, the ratio of organosulfur compounds to total polyphenolic compounds (e.g., the ratio of total organosulfur compounds to total bioflavonoids) is in the range of about 16:1 to about 1:30. For example, the ratio of organosulfur compounds to total polyphenolic compounds (e.g., the ratio of total organosulfur compounds to total bioflavonoids) can range from about 15:1 to about 1:30, or from about 14:1 to about 1:30, or from about 13:1 to about 1:30, or from about 12:1 to about 1:30, or from about 10:1 to about 1:30, or from about 16:1 to about 1:16. For example, the ratio of organosulfur compound to total polyphenolic compound (e.g., ratio of organosulfur compound to total bioflavonoid) may range from about 9:1 to about 1:25, or from about 8:1 to about 1:20, or from about 7:1 to about 1:15, or from about 6:1 to about 1:10, or from about 5:1 to about 1:8, or from about 4:1 to about 1:7, or from about 3:1 to about 1:6, or from about 2:1 to about 1:5, or from about 1:1 to about 1:4. For example, the ratio of organosulfur compounds to total polyphenolic compounds (e.g., the ratio of total organosulfur compounds to total bioflavonoids) can range from about 1:4 to about 1:8, or from about 1:1 to about 1:3, or from about 2:1 to about 1:4. For example, the ratio of organosulfur compounds to total polyphenolic compounds (e.g., the ratio of total organosulfur compounds to total bioflavonoids) may be about 1:6 or may be about 1:3. In a preferred embodiment, the organosulfur compound is a disulfide compound. In a preferred embodiment, the organosulfur compound is allicin and/or the polyphenolic compound is a bioflavonoid comprising naringin and neohesperidin.

In some embodiments, the organosulfur compound and the at least one polyphenol compound may be provided as a mixture. The mixture may be one as described in WO 2018/220340 A1, which is incorporated herein by reference. In the examples disclosed herein, the ratio of organosulfur compounds to total polyphenolic compounds in the mixture is 1:3, and the ratio of garlic powder to citrus extract is 93:7, which is referred to as "NXRH214" in the examples disclosed herein. In some examples disclosed herein, the ratio of organohalogen compound (e.g., bromoform) to powder mixture comprising organosulfur compound and polyphenol compound (e.g., NXRH214 powder) is 1:100 to 1:100000, more preferably 1:30000 to 1:100000, or 1:35000 to 1:83000.

-other additives

The composition may, for example, further comprise other animal feed supplements including, for example, vitamins, minerals, antibiotics, growth stimulators, and combinations thereof. For example, the composition may comprise other bioactive animal feed supplements, such as animal feed supplements suitable for reducing methane production/emissions and/or increasing nutrient availability to an animal. The vitamin may be any one or more of vitamin A, vitamin D, vitamin E, vitamin K, thiamine, riboflavin, pyridoxine, cyanocobalamine, carotenoids (including beta-carotene, zeaxanthin, lutein, and lycopene), niacin, folic acid, pantothenic acid, biotin, vitamin C, choline, inositol, and salts and derivatives thereof. The mineral may be any one or more of calcium, phosphorus, magnesium, iron, zinc, manganese, copper, cobalt, boron, iodine, sodium, potassium, molybdenum, selenium, chromium, fluorine and chlorine. The animal feed composition may, for example, comprise from about 0.001 wt% to about 5 wt% of each additional animal feed supplement, or from about 0.01 wt% to about 5 wt%, or from about 0.1 wt% to about 5 wt% of each additional animal feed supplement.

In addition to the organosulfur compound, the organohalogen compound, and optionally at least one polyphenolic compound, the composition may, for example, further comprise other components such as, for example, flavoring agents, colorants, stabilizers, antioxidants, buffers, emulsifiers, dispersants, thickeners, solubilizing agents, micronutrients (e.g., selenium), vitamins, other feed substances (e.g., carbohydrates such as sugar and starch), soluble and insoluble fibers, cellulose, lignocellulose, cereal grains, chaff, cereal grits, fruit and vegetable seeds, husks, pericarps, and the like.

-animal feed

Also disclosed herein is an animal feed comprising the composition described herein. Animal feed can be solid (e.g., powder, granules, pellets), semi-solid (e.g., gel, ointment, cream, paste) or liquid (e.g., solution, suspension, emulsion). Animal feed can be independently solid, semi-solid (e.g., gel, ointment, cream, paste) or liquid (e.g., solution, suspension, emulsion). For example, the animal feed may be all liquid, or all semi-solid, or all solid. Alternatively, the animal feed and the composition may each be in a different physical state. For example, the animal feed may be solid or semi-solid, while the composition may be liquid. The composition may be used, for example, in a "top-dressing" (top-additive) ruminant farm ration, or may be used to blend a total mixed ration. For example, the composition may be added to the drinking water of an animal. In certain embodiments, the composition may be added to the drinking water of an animal immediately prior to ingestion, for example, up to 1 hour prior to ingestion, or up to 30 minutes prior to ingestion, or up to 15 minutes prior to ingestion, or up to 5 minutes prior to ingestion. Three main types of animal feed include roughage, concentrate, and mixed feed. Generally, roughage comprises a higher percentage of crude fiber and a lower percentage of digestible nutrients than concentrate. For example, a roughage may be defined as comprising equal to or greater than 20 wt% of crude fiber and equal to or less than 60 wt% of total digestible nutrients. Roughages may include, for example, dry roughages (such as hay, straw, artificially dehydrated forage containing at least 90% by weight dry matter), silage (formed from green forage such as grass, alfalfa, sorghum, and corn, and stored in silos at 20% to 50% dry matter content), and pastures (such as green growing pastures that provide forage having a high moisture content and typically less than 30% dry matter). Two basic types of roughage include grasses and legumes. Grass fiber and dry matter are generally higher than legumes. Legume proteins, metabolizable energy, vitamins and minerals are generally higher. The concentrate comprises a relatively lower percentage of crude fiber and a higher percentage of digestible nutrients than the roughage. For example, concentrate may be defined as comprising less than 20% by weight crude fiber and greater than 60% by weight total digestible nutrients. Concentrate may include, for example, energy-rich grains and molasses. Corn, wheat, oats, barley and milo (sorghum grains) are energy rich grains comprising about 70 to 80 weight percent total digestible nutrients.

The mixed feed is typically a mixture of coarse and fine feed to provide a "complete" balanced ration, and the energy, protein or fiber may be high or low. The at least one organosulfur compound and the at least one polyphenolic compound (e.g., the at least one bioflavonoid) may be combined with the animal feed, for example, in various amounts, depending on the total amount of organohalogen compound(s), organosulfur compound(s) and optionally polyphenolic compound(s) (e.g., bioflavonoid) intended for administration to the animal.

The animal feed may, for example, comprise from about 0.0001 wt% to about 10 wt% of an organic sulfur compound (e.g., allicin) based on the total dry weight of the animal feed. The animal feed may, for example, comprise from about 0.3 wt% to about 10 wt% of an organic sulfur compound (e.g., allicin) based on the total dry weight of the animal feed. For example, the animal feed can comprise from about 0.001 wt% to about 9.5 wt%, or from about 0.005 wt% to about 9 wt%, or from about 0.01 wt% to about 8.5 wt%, or from about 0.05 wt% to about 8 wt%, or from about 0.1 wt% to about 7.5 wt%, or from about 0.9 wt% to about 7 wt%, or from about 1 wt% to about 6 wt%, or from about 1.5 wt% to about 5.5 wt%, or from about 2 wt% to about 5 wt%, or from about 2.5 wt% to about 4.5 wt%, or from about 3 wt% to about 4 wt% of an organic sulfur compound (e.g., allicin), based on the total dry weight of the animal feed. For example, the animal feed can comprise from about 0.4 wt% to about 9.5 wt%, or from about 0.5 wt% to about 9 wt%, or from about 0.6 wt% to about 8.5 wt%, or from about 0.7 wt% to about 8 wt%, or from about 0.8 wt% to about 7.5 wt%, or from about 0.9 wt% to about 7 wt%, or from about 1 wt% to about 6 wt%, or from about 1.5 wt% to about 5.5 wt%, or from about 2 wt% to about 5 wt%, or from about 2.5 wt% to about 4.5 wt%, or from about 3 wt% to about 4 wt% of an organic sulfur compound (e.g., allicin), based on the total dry weight of the animal feed. The concentration of total organic sulfur compounds (e.g., allicin) present in the animal feed supplements or animal feed compositions described herein generally exceeds the concentration of each organic halogen compound. As indicated above, the ratio of organohalogen compound to organosulfur compound in the animal feed can be about 1:10 to 1:3500, or 1:100 to 1:3500, or more preferably 1:1000 to 1:2500. Thus, in some embodiments, the animal feed can comprise from about 0.00015 wt% to about 0.01 wt% of an organic sulfur compound (e.g., allicin) based on the total dry weight of the animal feed.

The animal feed, if present, may, for example, comprise from about 0.0001% to about 10% by weight of total polyphenolic compounds (e.g., total bioflavonoids) based on the total dry weight of the animal feed. The animal feed may, for example, comprise from about 0.1% to about 10% by weight total polyphenolic compounds (e.g., total bioflavonoids) based on the total dry weight of the animal feed. For example, the animal feed can comprise from about 0.001 wt% to about 10 wt%, or from about 0.005 wt% to about 10 wt%, or from about 0.01 wt% to about 9.5 wt%, or from about 0.05 wt% to about 9 wt%, or from about 0.1 wt% to about 8.5 wt%, or from about 0.7 wt% to about 8 wt%, or from about 0.8 wt% to about 7.5 wt%, or from about 0.9 wt% to about 7 wt%, or from about 1 wt% to about 6 wt%, or from about 1.5 wt% to about 5.5 wt%, or from about 2 wt% to about 5 wt%, or from about 2.5 wt% to about 4.5 wt%, or from about 3 wt% to about 4 wt% of a total polyphenolic compound (e.g., a total bioflavanoid) based on the total dry weight of the animal feed. For example, the animal feed can comprise from about 0.2 wt% to about 10 wt%, or from about 0.3 wt% to about 10 wt%, or from about 0.4 wt% to about 9.5 wt%, or from about 0.5 wt% to about 9 wt%, or from about 0.6 wt% to about 8.5 wt%, or from about 0.7 wt% to about 8 wt%, or from about 0.8 wt% to about 7.5 wt%, or from about 0.9 wt% to about 7 wt%, or from about 1 wt% to about 6 wt%, or from about 1.5 wt% to about 5.5 wt%, or from about 2 wt% to about 5 wt%, or from about 2.5 wt% to about 4.5 wt%, or from about 3 wt% to about 4 wt% of a total polyphenolic compound (e.g., a total bioflavanoid) based on the total dry weight of the animal feed. The concentration of total organic sulfur compounds (e.g., allicin) present in the animal feed supplements or animal feed compositions described herein can exceed the concentration of total polyphenol compound(s).

-method of the present disclosure

Disclosed herein are methods of reducing methane, e.g., reducing methane production by an animal, comprising administering to the animal, more particularly a ruminant, a composition or animal feed described herein.

The compositions and methods described herein can, for example, reduce methane production and/or emissions by at least about 10% (as compared to methane production and/or emissions if the animal feed supplement is not consumed). For example, the animal feed supplement can reduce methane production and/or emissions by at least about 10%, or at least about 15%, or at least about 25%, or at least about 30%, or at least about 35%, or at least about 40%, or at least about 45%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%. The animal feed supplements described herein may, for example, reduce methane production and/or emissions by up to 100%. For example, an animal feed supplement may reduce methane production and/or emissions by up to about 99%, or up to about 98%, or up to about 97%, or up to about 96%, or up to about 95%, or up to about 90%, or up to about 85%, or up to about 80%, or up to about 75%, or up to about 70%. This can be measured, for example, by a Huo Enhai-mu (Hohenheim) gas test or by using a pressure gauge.

Also disclosed herein are methods of inhibiting one or more methanogens comprising administering to an animal, more particularly a ruminant, a composition or animal feed described herein. In some embodiments, the compositions and combinations disclosed herein can be used to reduce one or more methanogens selected from the group consisting of: methanobacterium (Methanobacterium formicicum), methanobacterium buchneri (Methanobacterium bryantii), methanobacterium rumens (Methanobrevibacter ruminantium), methanobacterium milhneri (Methanobrevibacter millerae), methanobacterium australis (Methanobrevibacter olleyae), methanobacterium motile (Methanomicrobium mobile), methanobacterium australis (Methanoculleus olentangyi), methanosarcina bardans (Methanosarcina barkeri), methanobacterium bortezomib (Methanobrevibacter boviskoreani), methanobacterium beijerinae (Methanobacterium beijingense), methanobacterium jersey (Methanoculleus marisnigri), methanobacterium breve (Methanoculleus bourgensis), methanosarcina mahogany (Methanosarcina mazei), methanobacterium gordonii (Methanobrevibacter gottschalkii), methanobacterium co-existence (Methanobrevibacter thaueri), methanobacterium smini (Methanobrevibacter smithii), methanobacterium stevensis (Methanosphaera stadtmanae), methanobacterium Wo Sishi (Methanobrevibacter woesei), and methanobacterium wovensis (Methanobrevibacter wolinii).

Also disclosed herein are methods of improving the metabolic efficiency of an animal comprising administering to the animal a composition or animal feed of the invention. Improvements in metabolic efficiency may result in increased production of animal products, such as one or more of meat, fat, wool (i.e., fiber), and milk. Thus, the compositions or methods of the invention may improve the production of meat and/or fat and/or hair and/or milk from animals.

The compositions, animal feeds and methods described herein can, for example, increase milk and/or meat and/or hair production by at least about 20% (as compared to if the composition or animal feed were not consumed). For example, the composition or animal feed may increase milk and/or meat and/or fat and/or hair production by at least about 25%, or at least about 30%, or at least about 35%, or at least about 40%, or at least about 45%, or at least about 50%. The compositions or animal feeds described herein may, for example, increase milk and/or meat and/or fat and/or hair production by up to 100%. For example, the composition or animal feed may increase milk and/or meat and/or fat and/or hair production by up to about 95%, or up to about 90%, or up to about 85%, or up to about 80%, or up to about 75%, or up to about 70%. This can be measured, for example, by the volume of milk produced per day or by the weight of the animal or by the weight of the hair and/or fat and/or meat produced.

For example, the compositions and animal feeds described herein can increase the efficiency of milk and/or meat and/or hair production by at least about 20% (as compared to the efficiency of milk and/or meat and/or fat and/or hair production if the composition or animal feed is not consumed). For example, the compositions or animal feeds described herein may increase the efficiency of milk and/or meat and/or fat and/or hair production by at least about 25%, or at least about 30%, or at least about 35%, or at least about 40%, or at least about 45%, or at least about 50%. The compositions or animal feeds described herein may, for example, increase the efficiency of milk and/or meat and/or fat and/or hair production by up to 100%. For example, the compositions or animal feeds described herein may increase the efficiency of milk and/or meat and/or fat and/or hair production by up to about 95%, or up to about 90%, or up to about 85%, or up to about 80%, or up to about 75%, or up to about 70%. Efficiency is related to the extent to which a particular biological process (e.g., milk, meat, fat, wool production) occurs per unit of nutrient consumed. This can be measured, for example, by the change in the volume of milk produced per day or the weight of the animal or the weight of hair or fat divided by the total nutrient consumed by the animal. For example, the compositions or animal feeds described herein can increase nutrient availability by at least about 20% (as compared to milk and/or meat and/or fat and/or hair production if the composition or animal feed is not consumed). For example, a composition or animal feed described herein can increase nutrient availability by at least about 25%, or at least about 30%, or at least about 35%, or at least about 40%, or at least about 45%, or at least about 50%. The compositions or animal feeds described herein may, for example, increase nutrient availability by up to 100%. For example, the compositions or animal feeds described herein can increase nutrient availability by up to about 95%, or up to about 90%, or up to about 85%, or up to about 80%, or up to about 75%, or up to about 70%. Nutrient availability refers to the amount of nutrient available to an animal for biological/metabolic function.

In some embodiments, the ruminant is a cow, goat, sheep, yak, deer, or antelope. In some embodiments, the ruminant is a cow, goat or sheep.

The composition or animal feed may be orally administered to an animal. In some embodiments, the composition or animal feed may be administered to the animal daily.

Improving the metabolic efficiency of ruminants

The present disclosure provides feed supplement formulations incorporating biologically or synthetically derived organic halogens, organic sulfur and polyphenolic compounds suitable for oral administration to ruminants to improve their metabolic efficiency, for reducing emitted methane and reducing excreted nitrogen, and for increasing valuable animal products such as meat, fat, fiber and milk.

The present invention is based on the following unexpected findings: when certain organohalogen compounds, organosulfur compounds and polyphenolic compounds are administered to ruminants to reduce methane emissions in the ruminant, these organohalogen compounds, organosulfur compounds and polyphenolic compounds also improve the metabolic efficiency of the ruminant, and also reduce urinary nitrogen excretion, and increase production of valuable animal products. Furthermore, when the organohalogen compound, organosulfur compound and polyphenolic compound are administered in certain combinations, there is a surprising increase in the reduction of both emitted methane and excreted nitrogen, and a surprising increase in the production of valuable animal products.

The present inventors have recognized that feed supplements from certain combinations comprising organohalogen-rich marine macroalgae and organosulfur-rich plants, as well as polyphenol-rich plants, provide unexpected and surprising improvements in metabolic efficiency.

The combination of organohalogen-rich marine macroalgae and organosulfur-rich plants and polyphenol-rich plants of the invention, when administered to ruminants, reduces methanogenesis and reduces ruminant methane production. The reduction in methanogenesis occurs through different modes including: reducing methanogenic organisms by limiting their growth or killing them; the methanogenic process is reduced by limiting or terminating enzymes involved in methanogenesis.

Methanogens identified in cattle, sheep and goats include methanobacterium formate (Methanobacterium formicicum), methanobacterium buchneri (Methanobacterium bryantii), methanobacterium rumens (Methanobrevibacter ruminantium), methanobacterium migrainae (Methanobrevibacter millerae), methanobacterium australis (Methanobrevibacter olleyae), methanobacterium motile (Methanomicrobium mobile), methanobacterium australis (Methanoculleus olentangyi), methanobacterium sarcina bardans (Methanosarcina barkeri), methanobacterium borgpi (Methanobrevibacter boviskoreani), methanobacterium beijing (Methanobacterium beijingense), methanobacterium jersey (Methanoculleus marisnigri), methanobacterium breveticus (Methanoculleus bourgensis), methanosarcina marxianus (Methanosarcina mazei), methanobacterium gossypii (Methanobrevibacter gottschalkii), methanobacterium co-existence (Methanobrevibacter thaueri), methanobacterium smithii (Methanobrevibacter smithii), methanobacterium stevens (Methanosphaera stadtmanae), methanobacterium Wo Sishi (Methanobrevibacter woesei) and methanobacterium wovens (Methanobrevibacter wolinii). Thus, in some embodiments, the compositions and combinations disclosed herein can be used to reduce one or more methanogens selected from the group consisting of: methanobacterium (Methanobacterium formicicum), methanobacterium buchneri (Methanobacterium bryantii), methanobacterium rumens (Methanobrevibacter ruminantium), methanobacterium migraines (Methanobrevibacter millerae), methanobacterium australis (Methanobrevibacter olleyae), methanobacterium motile (Methanomicrobium mobile), methanobacterium australis (Methanoculleus olentangyi), methanosarcina bardans (Methanosarcina barkeri), methanobacterium bortezomib (Methanobrevibacter boviskoreani), methanobacterium beijerinae (Methanobacterium beijingense), methanobacterium jersey (Methanoculleus marisnigri), methanobacterium breve (Methanoculleus bourgensis), methanosarcina mahogany (Methanosarcina mazei), methanobacterium gordonii (Methanobrevibacter gottschalkii), methanobacterium co-existence (Methanobrevibacter thaueri), methanobacterium smini (Methanobrevibacter smithii), methanobacterium stevens (Methanosphaera stadtmanae), methanobacterium Wo Sishi (Methanobrevibacter woesei) and methanobacterium vortiodans (Methanobrevibacter wolinii).

The combination of organohalogen-rich marine macroalgae and organosulfur-rich plants and polyphenol-rich plants of the invention has unexpected and surprising enhancements in reducing methane emissions and excreted urine nitrogen, as well as in increasing valuable animal products, possibly due to synergy between different modes of inhibition of methanogenesis including, for example:

organohalogen compounds from the species Asparagopsis (Asparagopsis) of marine macroalgae include organobromine, especially bromoform (CHBr) ₃ Bromoform), which inhibits the efficiency of methyltransferase by reacting with reduced vitamin B12 cofactor required for the second to last steps of methanogenesis, and also competitively inhibits methane production by acting as a terminal electron acceptor.

Organosulfur compounds from plants of the Allium (Allium) species include allicin and diallyl disulfide, which possess methanogenic activity due to oxidative interactions with important thiol-containing enzymes and by inhibiting the enzyme HMG-CoA reductase.

Polyphenolic compounds from plants of the Citrus (Citrus) species include the flavonoids neohesperidin and naringin, which have anti-methanogenic activity.

The inventors have recognized that due to the supplement of the present invention, unexpected and surprising improvements in metabolic efficiency may also be due to additional health benefits, including, for example: an anthelmintic effect resulting in a reduction of gastrointestinal parasites; antibacterial effects that lead to bacterial infections including, for example, reduced mastitis; and when administered to ruminants, providing supplemental trace minerals and vitamins present in the organohalogen-rich marine macroalgae and organosulfur-rich plants and polyphenol-rich plant combinations of the invention.

The combination of organohalogen-rich marine macroalgae and organosulfur-rich plants of the invention, and polyphenol-rich plants, can be administered as a feed supplement in certain combinations and in certain proportions.

The combination of organohalogen-rich marine macroalgae and organosulfur-rich plants and polyphenol-rich plants of the invention can be administered as a separate feed supplement or combined into a mixed composition feed supplement.

Use of the same

The compositions or animal feed supplements described herein (including all embodiments and combinations of embodiments) can be used to reduce methane production and/or emissions by an animal, reduce nitrogen excretion by an animal, increase the availability of nutrients to an animal, and/or increase the animal's valuable nitrogen-and carbon-enriched animal products.

In certain embodiments, the animal is a ruminant. Ruminants include animals selected from members of the sub-orders ruminant (Ruminaria) and foot-callus (Tylopoda), and include domesticated ruminants: for example, cattle (e.g., cows), goats, sheep, buffalo, yaks, deer or antelope.

In particular, the compositions or feed supplements of the present invention, when administered to ruminants in an effective amount, result in reduced methane production by the ruminants, which would otherwise be vented to the atmosphere primarily through mouth and nostril exhalation gases, and represent an energy loss of 2% to 12% of the total energy intake from the feed.

Methane is a greenhouse gas with a global warming potential 28 times that of carbon dioxide. Intestinal methane is a byproduct of ruminant digestion and is produced by complex microflora including ciliate protozoa, bacteria, archaebacteria, and anaerobic fungi through a process known as methanogenesis. Cattle produce about 7 and 9 times as much methane as sheep and goats, respectively. Intestinal methane is produced mainly in the rumen (87% -90%) and to a lesser extent (13% -10%) in the large intestine.

The compositions and feed supplements of the present invention result in the transfer of metabolic energy from methane production and direct it to anabolic growth processes. Thus, the feed supplement results in an increase in the live weight of ruminants, which is determined by: directly weighing the animal mass; computed Tomography (CT) scans to measure empty body mass (total mass minus intestinal content); carcass quality and composition analysis (lean, fat and major fat distribution; and organ including liver changes).

Examples of valuable nitrogen-and carbon-enriched animal products are tissue-based commodities and include, for example: meat; viscera; and leather.

Another example of valuable nitrogen-and carbon-rich animal products are secretion-based commodities and those commodity products, including for example: milk; full fat milk; milk powder; cream; ice cream; cheese; and yogurt.

Another example of a valuable nitrogen-and carbon-enriched animal product is a fiber-based commodity and includes, for example: a hair; angle (horn); and deer horn (antler).

The compositions and feed supplements of the present invention result in an unexpected and surprising improvement in metabolic efficiency, which may result in a reduction of excreted urinary nitrogen that is deposited in urine plaques on pastures after urination. When the excess excreted nitrogen in the urine plaques is greater than that required for optimal pasture plant efficiency, the excess nitrogen goes through nitrate (NO ₃ ^- ) Leaching and ammonia (NH) ₃ ) Dinitrogen monoxide (N) ₂ O) and nitrogen (N) ₂ ) And volatilized and lost. Nitrous oxide is particularly harmful to the atmosphere as a greenhouse gas and has a global warming potential 298 times that of carbon dioxide. Loss of nitrogen into groundwater can lead to uncontrolled growth of aquatic microbiota, thereby destroying the ecosystem, leading to massive proliferation of toxic algae and eutrophication of the water body.

Method of manufacture

The compositions or animal feed supplements described herein may be prepared by combining one or more organohalogen compounds and one or more organosulfur compounds and one or more polyphenolic compounds.

The organohalogen compounds can be synthesized or extracted from a suitable biological source and used in raw or processed form. For example, the organohalogen compounds include: CH (CH) ₃ Cl；CH ₃ Br；CH ₃ I；CH ₂ Cl ₂ ；CH ₂ Br ₂ ；CH ₂ I ₂ ；CHCl ₃ ；CHBr ₃ ；CHI ₃ ；CCl ₄ ；CBr ₄ ；CH ₂ ClBr；CH ₂ ClI；CH ₂ BrI；CHBr ₂ Cl；CHBrI ₂ ；CHBrClI；CHBr ₂ I；CHBrCl ₂ ；CH ₃ CH ₂ Br；CH ₃ CH ₂ I；CH ₃ CH ₂ CH ₂ I；CH ₃ (CH ₂ ) ₃ I；CH ₃ (CH ₂ ) ₄ Br；CH ₃ (CH ₂ ) ₄ I；(CH3) ₂ CHI；CH ₃ CH ₂ CH(CH ₃ )I；(CH ₃ ) ₂ CHCH ₂ I；BrCH ₂ CH ₂ Br；ClCH＝CCl ₂ The method comprises the steps of carrying out a first treatment on the surface of the And CH (CH) ₃ CH ₂ CH ₂ CH ₂ I。

The biological source of the organohalogen compound is an organohalogen-rich marine macroalgae. For example, the organic halogen-rich marine macroalgae comprises at least one marine macroalgae selected from the group consisting of: radix seu radix Ophiopogonii (Asparagopsis armata); taxus-like sea-door winter (Asparagopsis taxiformis); a Dictyota (Dictyota) species; a sphingosine (oendonium) species; ulva (Ulva) species; and cladophora (Cladophora patentiramea).

The organosulfur compounds can be synthesized or extracted from suitable biological sources and used in raw or processed form. The organic sulfur compounds include: organic sulfur secondary metabolites; allicin (C) ₆ H ₁₀ S ₂ O); diallyl sulfide (C) ₆ H ₁₀ S) S; diallyl disulfide (C) ₆ H ₁₀ S ₂ ) The method comprises the steps of carrying out a first treatment on the surface of the And allyl mercaptan (C) ₃ H ₆ S)。

The biological source of the organosulfur compounds is an organosulfur-rich plant. For example, organic sulfur-rich plants include: allium species; sativum (garlic); ampeloprasum (leek); cepa (onion and green onion). The one or more organosulfur compounds can be obtained from one or more parts of a plant, including, for example: leaves; stems; bark; root; bulbs; flower; fruit; and seeds.

The polyphenol compounds may be synthesized or extracted from a suitable biological source and used in raw or processed form. For example, polyphenolic compounds include bioflavonoids and phenolic compounds. The term phenolic compound refers to a class of chemical compounds that contain hydroxyl groups (-OH) directly bonded to aromatic hydrocarbon groups. The phenolic compounds described herein may include bioflavonoids, non-bioflavonoid phenolic compounds, or combinations thereof. The at least one polyphenolic compound may, for example, comprise at least one bioflavonoid. The term bioflavonoids refers to a class of plant and fungal secondary metabolites and has the general structure of a 15 carbon backbone consisting of two benzene rings (A and B) and a heterocycle (C), sometimes abbreviated as C ₆ -C ₃ -C ₆ . The term bioflavonoids includes anthocyanins (including flavones and flavonols), flavanones, flavanonols, flavans and anthocyanidins. The term bioflavonoids also includes compounds having a flavone backbone (2-phenyl-1, 4-benzopyrone), an isoflavane backbone (3-phenylchromen-4-one) or a neoflavane backbone (4-phenylcoumarin). Thus, the term polyphenol compound includes, but is not limited to: -a flower flavin; flavanones (including flavanone glycosides); flavonols; a flavanonol; a flavan; isoflavone; anthocyanin; procyanidins; phenolic acid; hydroxycinnamic acid; coumarin; stilbenes; anthraquinone; lignans; lignin; tannin; a polyphenol protein; catechin; rutin; robinia pseudoacacia; genistein; kaempferol; gallocatechin; catechin gallate; epicatechin; epigallocatechin; epicatechin gallate; quercetin; catechin; gallocatechin gallate; epicatechin; epigallocatechin; epicatechin gallate; epigallocatechin gallate; kaempferol; quercetin; naringin; neohesperidin; eriocitrin; naringin; naringenin; hesperidin; rhus verniciflua Stokes glycoside; myrosin; melissa glucoside; hesperetin; poncirin; epicatechin; gallocatechin; epigallocatechin; coumaric acid; cinnamic acid; gallic acid; ellagic acid; protocatechuic acid; chlorogenic acid; caffeic acid; ferulic acid; anan (safety) Punicalagin; granatum tann.

The biological source of polyphenol compounds is a polyphenol-rich plant. For example, polyphenol-enriched plants include: allium species; a Brassica (Brassica) species; a camellia (Camelia) species; a Capsicum (Capsicum) species; citrus (Citrus) species; cucumber (Cucumis) species; malus (Malus) species; a Musa (Musa) species; a Phaseolus (Phaseolus) species; a prune (prune) species; a Punica (Punica) species; pear (Pyrus) species; solanum (Solanum) species; vaccinium species. The one or more polyphenolic compounds may be obtained from one or more parts of a plant, including, for example: leaves; stems; bark; root; bulbs; flower; fruit; and seeds.

The compositions or animal feed supplements described herein may be prepared by combining one or more organohalogen-rich marine macroalgae and one or more organosulfur-rich plant compounds and one or more polyphenols-rich plants.

The components are combined in the appropriate amounts to obtain a composition having the desired amounts of the components. Each component may be combined with one or more other components in any order and combination suitable to obtain the desired product. For example, each component may be combined by mixing or blending. For example, the one or more organic halogen compounds and the one or more organic sulfur compounds and the one or more polyphenolic compounds may be combined with the animal feed by placing the one or more organic halogen compounds and the one or more organic sulfur compounds and the one or more polyphenolic compounds on top of the animal feed (Shi Dingfei).

The composition may be prepared in a dry solid form, such as a powder form, and subjected to further processing steps depending on the type of formulation for which the final product is intended. The method may further comprise a shaping step wherein the mixture is molded, pressed, spray dried or otherwise shaped into a shape (e.g., bar, sphere, pellet, cluster, tablet), preferably of a size and/or texture suitable for consumption by animals of the type described herein. The method may include containing the animal feed or animal feed supplement in a specific delivery device such as a syringe. The method may include preparing an animal feed supplement or animal feed into a pellet that is expected to reside in the stomach of an animal (e.g., the rumen of a ruminant).

Administration of methanogenesis inhibitors

Also disclosed herein are methods of stepwise administration of a methanogenesis inhibitor feed supplement incorporating biologically derived organohalogen compounds and/or organosulfur compounds and/or polyphenolic compounds, suitable for oral administration to ruminants to improve their metabolic efficiency, for reducing emitted methane and reducing excreted nitrogen, and for increasing valuable animal products such as meat, fat, fiber and milk.

The term "step-wise" as used herein means that at least one dose of an effective amount of at least one methanogenesis inhibitor is administered, and optionally at least one continuous dose of an effective amount of at least one methanogenesis inhibitor is administered after some effective time intervals.

The present disclosure is based on the unexpected discovery that the feed supplement, when administered stepwise to the animal in an effective dosage amount and for an effective time interval, provides surprising economic benefits by improving metabolic efficiency, reducing methane emissions and reducing nitrogen excretion, and increasing valuable animal products such as meat, fat, fiber, and milk.

Inhibition of methanogenesis occurs through different modes of action including, for example: reducing the methanogenesis process; by limiting or stopping enzymes involved in methanogenesis; or by limiting the growth of methanogenic organisms or killing them.

Methanogenesis inhibitors include, for example: an organohalogen compound; CH (CH) ₃ Cl；CH ₃ Br；CH ₃ I；CH ₂ Cl ₂ ；CH ₂ Br ₂ ；CH ₂ I ₂ ；CHCl ₃ ；CHBr ₃ ；CHI ₃ ；CCl ₄ ；CBr ₄ ；CH ₂ ClBr；CH ₂ ClI；CH ₂ BrI；CHBr ₂ Cl；CHBrI ₂ ；CHBrClI；CHBr ₂ I；CHBrCl ₂ ；CH ₃ CH ₂ Br；CH ₃ CH ₂ I；CH ₃ CH ₂ CH ₂ I；CH ₃ (CH ₂ ) ₃ I；CH ₃ (CH ₂ ) ₄ Br；CH ₃ (CH ₂ ) ₄ I；(CH3) ₂ CHI；CH ₃ CH ₂ CH(CH ₃ )I；(CH ₃ ) ₂ CHCH ₂ I；BrCH ₂ CH ₂ Br；ClCH＝CCl ₂ ；CH ₃ CH ₂ CH ₂ CH ₂ I, a step of I; an organic sulfur compound; organic sulfur secondary metabolites; allicin (C) ₆ H ₁₀ S ₂ O); diallyl sulfide (C) ₆ H ₁₀ S) S; diallyl disulfide (C) ₆ H ₁₀ S ₂ ) The method comprises the steps of carrying out a first treatment on the surface of the Allyl mercaptan (C) ₃ H ₆ S) S; a polyphenol compound; flavonoids; bioflavonoids; a non-bioflavonoid; -a flower flavin; flavone; flavonols; flavanones; a flavanonol; a flavan; anthocyanin; isoflavane; new flavanthamine; isoflavone; procyanidins; phenolic acid; hydroxycinnamic acid; coumarin; stilbenes; anthraquinone; lignans; lignin; tannin; a polyphenol protein; catechin; rutin; robinia pseudoacacia; genistein; kaempferol; gallocatechin; catechin gallate; epicatechin; epigallocatechin; epicatechin gallate; quercetin; catechin; gallocatechin gallate; epicatechin; epigallocatechin; epicatechin gallate; epigallocatechin gallate; kaempferol; quercetin; naringin; neohesperidin; eriocitrin; naringin; naringenin; hesperidin; rhus verniciflua Stokes glycoside; myrosin; melissa glucoside; hesperetin; poncirin; epicatechin; gallocatechin; epigallocatechin; coumaric acid; cinnamic acid; gallic acid; ellagic acid; protocatechuic acid; chlorogenic acid; caffeic acid; ferulic acid; punicalagin; pomegranate rind tannagin.

The biological source of the organohalogen compound is an organohalogen-rich marine macroalgae. For example, the organohalogen-rich marine macroalgae comprises at least one marine macroalgae selected from the group consisting of: radix seu radix Ophiopogonii (Asparagopsis armata); taxus-like sea-door winter (Asparagopsis taxiformis); a Dictyota (Dictyota) species; a sphingosine (oendonium) species; ulva (Ulva) species; and cladophora (Cladophora patentiramea). The organohalogen compounds, organobromine compounds, and bromoforms disclosed herein can also be synthetic.

The biological source of the organosulfur compounds is an organosulfur-rich plant. For example, organic sulfur-rich plants include: allium species; sativum (garlic); ampeloprasum (leek); cepa (onion and green onion). The one or more organosulfur compounds can be obtained from one or more parts of a plant, including, for example: leaves; stems; bark; root; bulbs; flower; fruit; and seeds. The organosulfur compounds disclosed herein can also be synthetic.

The biological source of polyphenol compounds is a polyphenol-rich plant. For example, polyphenol-enriched plants include: allium species; a Brassica (Brassica) species; a camellia (Camelia) species; a Capsicum (Capsicum) species; citrus (Citrus) species; cucumber (Cucumis) species; malus (Malus) species; a Musa (Musa) species; a Phaseolus (Phaseolus) species; a prune (prune) species; a Punica (Punica) species; pear (Pyrus) species; solanum (Solanum) species; vaccinium species. The one or more polyphenolic compounds may be obtained from one or more parts of a plant, including, for example: leaves; stems; bark; root; bulbs; flower; fruit; and seeds. The polyphenol compounds disclosed herein may also be synthetic.

The inventors have appreciated that by optimizing the stepwise administration by optimizing both the amount of effective dose and the effective time interval between successive doses:

1. maximizing the increase in valuable animal products in the ruminant animals, contributing to economic benefits; and

2. preventing overfeeding or other overfeeding, which reduces the cost of the feed supplement, contributes to economic benefit; and

3. preventing overfeeding or other overfeeding, which prevents counter productivity of the feed supplement in the ruminant animal, contributes to economic benefit.

The present inventors have recognized the unexpected and surprising economic benefits of the gradual administration of a methanogenesis inhibitor feed supplement comprising certain combinations of organohalogen-rich marine macroalgae and/or organosulfur-rich plants and/or polyphenol-rich plants by careful control.

The combination of organohalogen-rich marine macroalgae and/or organosulfur-rich plants and/or polyphenol-rich plants, when administered to ruminants, reduces methanogenesis and reduces ruminant methane production. The reduction in methanogenesis occurs through different modes including: reducing methanogenic organisms by limiting their growth or killing them; the methanogenic process is reduced by limiting or terminating enzymes involved in methanogenesis.

The progressive administration of certain combinations of organohalogen-rich marine macroalgae and organosulfur-rich plants and polyphenol-rich plants of the invention to ruminants has unexpected and surprising enhancements in reducing methane emissions and excreted urine nitrogen, as well as in increasing valuable animal products, possibly due to synergy between different modes of inhibition of methanogenesis including, for example:

1. organohalogen compounds from the species Asparagopsis (Asparagopsis) of marine macroalgae include organobromine, especially bromoform (CHBr) ₃ Bromoform), which inhibits the efficiency of methyltransferase by reacting with reduced vitamin B12 cofactor required for the second to last steps of methanogenesis, and also competitively inhibits methane production by acting as a terminal electron acceptor.

2. Organosulfur compounds from plants of the Allium (Allium) species include allicin and diallyl disulfide, which possess methanogenic activity due to oxidative interactions with important thiol-containing enzymes and by inhibiting the enzyme HMG-CoA reductase.

3. Polyphenolic compounds from plants of the Citrus (Citrus) species include the flavonoids neohesperidin and naringin, which have anti-methanogenic activity.

The inventors have recognized that due to the stepwise administration of the methanogenesis inhibitor feed supplement or composition of the present invention, the unexpected and surprising improvement in metabolic efficiency may also be due to additional health benefits, including, for example: an anthelmintic effect resulting in a reduction of gastrointestinal parasites; antibacterial effects that lead to bacterial infections including, for example, reduced mastitis; and when administered to ruminants, providing supplemental trace minerals and vitamins present in the organohalogen-rich marine macroalgae and organosulfur-rich plants and polyphenol-rich plant combinations of the invention.

The progressive administration of organohalogen-rich marine macroalgae and/or organosulfur-rich plants and/or polyphenol-rich plants of the invention can be applied as a feed supplement in certain combinations and in certain proportions.

The progressive administration of the organohalogen-rich marine macroalgae and/or the combination of organosulfur-rich plants and polyphenol-rich plants of the invention can be administered as a separate feed supplement or combined into a mixed composition feed supplement (as a composition described herein).

Use of the same

Progressive administration of the methanogenesis inhibitor animal feed supplements described herein (including all embodiments and combinations of embodiments) can be used to optimize the feed supplement administration to animals to reduce methane production and/or emissions from the animals, reduce nitrogen excretion from the animals, increase the availability of nutrients to the animals, and/or increase the animal's valuable nitrogen-and carbon-enriched animal products.

In particular, progressive administration of the methanogenesis inhibitor feed supplement or composition of the present invention when administered to ruminants results in reduced methane production by ruminants, which would otherwise be discharged into the atmosphere primarily through mouth and nostril exhalation gases, and represents an energy loss of 2% to 12% of the total energy intake from the feed.

The progressive administration of the methanogenesis inhibitor feed supplement of the present invention results in the transfer of metabolic energy from methane production and directs it to anabolic growth processes. Thus, the composition or feed supplement results in an increase in the live weight of the ruminant, as determined by: directly weighing the animal mass; computed Tomography (CT) scans to measure empty body mass (total mass minus intestinal content); carcass quality and composition analysis (lean, fat and major fat distribution; and organ including liver changes).

Another example of a valuable nitrogen-and carbon-enriched animal product is a fiber-based commodity and includes, for example: a hair; a corner; and deer horn.

The progressive administration of the methanogenesis inhibitor feed supplement of the present invention results in an unexpected and surprising improvement in metabolic efficiency, which may result in a reduction of excreted urinary nitrogen that is deposited in urinary plaques on pastures after urination. Overexcretion of nitrogen in urine plaques is greater than that required for optimal pasture plant efficiencyThe amount of nitrogen goes through nitrate (NO ₃ ^- ) Leaching and ammonia (NH) ₃ ) Dinitrogen monoxide (N) ₂ O) and nitrogen (N) ₂ ) And volatilized and lost. Nitrous oxide is particularly harmful to the atmosphere as a greenhouse gas and has a global warming potential 298 times that of carbon dioxide. Loss of nitrogen into groundwater can lead to uncontrolled growth of aquatic microbiota, thereby destroying the ecosystem, leading to massive proliferation of toxic algae and eutrophication of the water body.

Method of stepwise administration

The progressive administration of the compositions or methanogenesis inhibitor feed supplements described herein may be performed by administering at least one dose of an effective amount of at least one methanogenesis inhibitor and optionally administering at least one continuous dose of an effective amount of at least one methanogenesis inhibitor after some effective time intervals.

It will be appreciated that successive doses of the methanogenesis inhibitor feed supplement at time intervals constitute a step-wise administration. Furthermore, the amount of consecutive doses, the time interval between doses, and the methanogenic inhibitor may be the same from dose to dose, from time interval to time interval, and between methanogenic inhibitor and methanogenic inhibitor, or they may be different dose amounts and/or different time intervals, and/or different methanogenic inhibitors.

The animal feed supplements or compositions described herein may be prepared by combining one or more organohalogen-rich marine macroalgae and one or more organosulfur-rich plant compounds and one or more polyphenols-rich plants.

A methanogenesis inhibitor feed supplement may be prepared to aid in the stepwise administration in a form comprising: the solid form, for example a powder form, is dried and subjected to further processing steps depending on the type of formulation for which the final product is intended. The method may further comprise a shaping step wherein the mixture is molded, pressed, spray dried or otherwise shaped into a shape (e.g., bar, sphere, pellet, cluster, tablet), preferably of a size and/or texture suitable for consumption by animals of the type described herein. The method may include containing the animal feed or the animal methanogenesis inhibitor feed supplement in a specific delivery device such as a syringe. The method may include preparing the composition or animal feed into a pellet that is expected to reside in the stomach of an animal (e.g., the rumen of a ruminant).

The methanogenesis inhibitor feed supplement may be administered, for example, stepwise in dosage amounts based on weight percent of ruminants selected from the group consisting of: 0.1%;0.2%;0.3%;0.4%;0.5%;0.6%;0.7%;0.8%;0.9%;1.0%;1.1%;1.2%;1.3%;1.4%;1.5%;2.0%;2.5%;3.0%;3.5%;4.0%;4.5%;5.0%; and 10%.

The methanogenesis inhibitor feed supplement may be administered, for example, stepwise in a dosage amount based on the weight of feed consumed by the ruminant, selected from the group consisting of: 0.1%;0.2%;0.3%;0.4%;0.5%;0.6%;0.7%;0.8%;0.9%;1.0%;1.1%;1.2%;1.3%;1.4%;1.5%;2.0%;2.5%;3.0%;3.5%;4.0%;4.5%;5.0%; and 10%.

Gradual administration of the methanogenesis inhibitor feed supplement may have, for example, a time interval between at least one consecutive dose selected from the group consisting of: 1 minute; 1 hour; 1 day; for 2 days; 3 days; 4 days; for 5 days; for 6 days; 7 days; for 10 days; 2 weeks; 3 weeks; 4 weeks; for 6 weeks; 2 months; 3 months; 4 months; 6 months; 9 months; and 12 months.

All uses and methods described herein are considered purely non-therapeutic.

The invention will now be described by reference only to the following non-limiting examples.

EXAMPLE 1 inhibition of methane production by methanogenic archaea Methanococcus marinus (Methanococcus maripaludis)

i) Bromoform and allicin

The purpose of this experiment was to determine if bromoform and allicin were synergistic in their ability to inhibit methane production by the methanogenic archaebacteria methanococcus marinus (Methanococcus maripaludis).

For this experiment, 100mM bromoform was prepared by adding 8.75. Mu.l bromoform to 991. Mu.l DMSO. It was diluted 10-fold to produce a 10mM solution and further diluted to produce 0.12mM and 0.156mM stock solutions. An allicin stock solution was prepared by adding 48.6 μl of allicin to 951.4 μl of DMSO to prepare a 300mM stock solution, and adding 32.5 μl of allicin to 967.5 μl of DMSO to prepare a 200mM stock solution. Experiments were performed by adding 5ml of M141 medium (https:// www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium 141. Pdf) to a screw cap Hungate tube followed by 5. Mu.l allicin and/or 5. Mu.l bromoform or 10. Mu.l DMSO as follows:

tube 1: DMSO (10 μl)

Tube 2: bromoform (5. Mu.l 0.120 mM) (final 120 nM) +5. Mu.l DMSO

Tube 3: bromoform (5. Mu.l 0.156 mM) (final 156 nM) +5. Mu.l DMSO

Tube 4: allicin (5. Mu.l 200 mM) (final 200. Mu.M) +5. Mu.l DMSO

Tube 5: allicin (5. Mu.l 300 mM) (final 300. Mu.M) +5. Mu.l DMSO

Tube 6: bromoform (5. Mu.l 0.120 mM) (final 120 nM) +allicin (5. Mu.l 200 mM) (final 200. Mu.M)

Tube 7: bromoform (5. Mu.l 0.120 mM) (final 120 nM) +allicin (5. Mu.l 300 mM) (final 300. Mu.M)

Tube 8: bromoform (5. Mu.l 0.156 mM) (final 156 nM) +allicin (5. Mu.l 200 mM) (final 200. Mu.M)

Tube 9: bromoform (5. Mu.l 0.156 mM) (final 156 nM) +allicin (5. Mu.l 300 mM) (final 300. Mu.M)

After the addition of the test substances, 500. Mu.l of an overnight Methanococcus marinus (M.maripaludis) culture was added to each reaction tube. 80% H for each tube ₂ /20％ CO ₂ Aerated to 240kPa and incubated at 37 ℃ for 24 hours.

After 24 hours incubation, the pressure inside the tube was measured using a pressure gauge. Assuming 5 moles of H are consumed ₂ /CO ₂ Generating 1 mole of CH ₄ The observed pressure drop is used to calculate the amount of methane for the control and test reactions, and thus the percent inhibition.

ii) bromoform and powder comprising organosulfur and polyphenol (NXRH 214 powder)

The purpose of this experiment was to determine if bromoform and powders comprising organosulfur and polyphenol were synergistic in their ability to inhibit methane production by methanogenic archaebacteria methanococcus maritimus (Methanococcus maripaludis).

For this experiment, 100mM bromoform was prepared by adding 8.75. Mu.l bromoform to 991. Mu.l DMSO. It was diluted 10-fold to produce a 10mM solution and further diluted to produce 0.10mM and 0.156mM stock solutions. Samples were prepared by adding 245mg of NXRH 14 powder to 35ml of M141 medium (https:// www.dsmz.de/microorganisms/media/pdf/DSMZ_Medium 141. Pdf) to produce 7mg/ml stock solution. The N XRH214 powder is garlic powder (allicin) and citrus extract (polyphenol flavonoid mixture) in a ratio of 93:7, wherein the flavonoid mixture mainly comprises naringin and neohesperidin.

The reaction tube was set as follows:

tube 1:0ml of NXRH 14 (no NXRH 214) +5ml of M141+5. Mu.l of DMSO

Tube 2:1ml of NXRH 14 (1.4. Mu.g/ml NXRH 214) +5ml of M141+5. Mu.l DMSO tube 3:1.5ml NXRH 14 (2.8. Mu.g/ml NXRH 214) +3.5ml M141+5. Mu.l DMSO tube 4:0ml NXRH 14 (no NXRH 214) +5ml M141+5. Mu.l 0.1mM bromoform (100 nM bromoform)

Tube 5:1ml of NXRH 14 (1.4. Mu.g/ml NXRH 214) +5ml of M141+5. Mu.l of 0.1mM bromoform (100 nM bromoform)

Tube 6:1.5ml NXRH 14 (2.8. Mu.g/ml NXRH 214) +3.5ml M141+5. Mu.l 0.1mM bromoform (100 nM bromoform)

Tube 7:0ml NXRH 14 (no NXRH 214) +5ml M141+5. Mu.l 0.156mM bromoform (156 nM bromoform)

Tube 8:1ml of NXRH 14 (1.4. Mu.g/ml NXRH 214) +5ml of M141+5. Mu.l of 0.156mM bromoform (156 nM bromoform)

Tube 9:1.5ml NXRH 14 (2.8. Mu.g/ml NXRH 214) +3.5ml M141+5. Mu.l 0.156mM bromoform (156 nM bromoform)

After 24 hours incubation, the pressure inside the tube was measured using a pressure gauge. Assuming 5 moles of H are consumed ₂ /CO ₂ Generating 1 mole of CH ₄ The observed pressure drop is used to calculate the amount of control and test reaction methane, and thus the percent inhibition.

Experimental results

i) The experimental results of bromoform and allicin and ii) bromoform and powders comprising organosulfur and polyphenol (i.e. NXRH214 powder) are shown in fig. 1 and 2, respectively, and in tables 1 and 2 below.

Table 1: methane inhibition was achieved with various compositions, including compositions comprising bromoform and allicin.

	Inhibiting gas%
		DMSO	0
DMSO/bromoform 120nM	0
		DMSO/bromoform 156nM	0
DMSO/allicin 200. Mu.M	0
		DMSO/allicin 300. Mu.M	19
Bromoform 120 nM/allicin 300. Mu.M	75
		Bromoform 156 nM/allicin 200. Mu.M	39
Bromoform 156 nM/allicin 300. Mu.M	99

The results show a significant synergy of the compositions comprising bromoform and allicin, which significantly exceeded the% gas inhibition exhibited by either component alone.

Table 2: methane inhibition was achieved with various compositions, including compositions comprising bromoform and powders comprising organosulfur and polyphenol (i.e., NXRH214 powder), which is a mixture of allicin and bioflavonoids.

Sample of	Inhibiting gas%
		DMSO	0
Bromoform 100nM	0
		Bromoform 156nM	0.5
NXRH214 powder 1.4mg/ml	2.2
		Bromoform 100nM+NXRH214 powder 1.4mg/ml	44.8
Bromine simulation 156nM+NXRH214 powder 1.4mg/ml	100
		NXRH214 powder 2.1mg/ml	88.5
Bromoform 100nM+NXRH214 powder 2.1mg/ml	96.2
		Bromine simulation 156nM+NXRH214 powder 2.1mg/ml	100

The results also show a significant synergy of the composition comprising bromoform and the powder comprising organosulfur and polyphenol (i.e., the NXRH214 powder), which significantly exceeded the% gas inhibition exhibited by either component in the presence of DMSO. The NXRH214 powder comprises allicin and polyphenolic compounds, more specifically the bioflavonoids naringin and neohesperidin. Thus, compositions comprising bromoform, organosulfur compounds, and polyphenols may also be used to effectively inhibit methane production.

Conclusion(s)

The above data clearly show a high% inhibition of methanogens when organohalogen (i.e. bromoform) is combined with organosulfur (i.e. allicin) alone or with polyphenol (i.e. bioflavonoids from citrus extracts).

Example 2

From Mootral, switzerland ^TM SA development and sales feed supplement Mootral ^TM And marine macroalgae, asparagopsis (Asparagopsis armata) are combined in various proportions and given stepwise in various dosage amounts and at various time intervals to a fully grass fed sheep. The methane, blood, body and stool output were measured and compared to control animals that did not receive the supplement.

The disclosure may also be described by the following paragraphs

A. A method of reducing nitrogen excretion by a ruminant and/or reducing methane emissions by a ruminant and/or increasing nitrogen-rich and carbon-rich materials in a ruminant, the method comprising the step of administering to the ruminant an effective amount of at least one type of methanogenesis inhibitor.

B. A method according to paragraph a, wherein the methanogenesis inhibitor is selected from the group consisting of: an organohalogen compound; marine macroalgae enriched in organic halogen; an organic sulfur compound; plants rich in organic sulfur; a polyphenol compound; and polyphenol-enriched plants.

C. A method according to paragraph B, wherein the organohalogen compound is selected from: CH (CH) ₃ Cl；CH ₃ Br；CH ₃ I；CH ₂ Cl ₂ ；CH ₂ Br ₂ ；CH ₂ I ₂ ；CHCl ₃ ；CHBr ₃ ；CHI ₃ ；CCl ₄ ；CBr ₄ ；CH ₂ ClBr；CH ₂ ClI；CH ₂ BrI；CHBr ₂ Cl；CHBrI ₂ ；CHBrClI；CHBr ₂ I；CHBrCl ₂ ；CH ₃ CH ₂ Br；CH ₃ CH ₂ I；CH ₃ CH ₂ CH ₂ I；CH ₃ (CH ₂ ) ₃ I；CH ₃ (CH ₂ ) ₄ Br；CH ₃ (CH ₂ ) ₄ I；(CH3) ₂ CHI；CH ₃ CH ₂ CH(CH ₃ )I；(CH ₃ ) ₂ CHCH ₂ I；BrCH ₂ CH ₂ Br；ClCH＝CCl ₂ The method comprises the steps of carrying out a first treatment on the surface of the And CH (CH) ₃ CH ₂ CH ₂ CH ₂ I。

D. A method according to paragraph B, wherein the organohalogen rich marine macroalgae is selected from the group consisting of: radix seu radix Ophiopogonii (Asparagopsis armata); taxus-like sea-door winter (Asparagopsis taxiformis); a Dictyota (Dictyota) species; a sphingosine (oendonium) species; ulva (Ulva) species; and cladophora (Cladophora patentiramea).

E. A method according to paragraph B, wherein the organosulfur compound is selected from the group consisting of: organic sulfur secondary metabolites; allicin (C) ₆ H ₁₀ S ₂ O); diallyl sulfide (C) ₆ H ₁₀ S) S; diallyl disulfide (C) ₆ H ₁₀ S ₂ ) The method comprises the steps of carrying out a first treatment on the surface of the And allyl mercaptan (C) ₃ H ₆ S)。

F. A method according to paragraph B, wherein the organosulfur-rich plant is selected from the Allium species: garlic (Allium sativum); green Chinese onion (Allium ampeloprasum); and onion (Allium cepa).

G. A method according to paragraph B, wherein the polyphenolic compound is selected from the group consisting of: flavonoids; bioflavonoids; non-bioflavonoids. The at least one polyphenolic compound may, for example, comprise at least one bioflavonoid; -a flower flavin; flavone; flavonols; flavanones; a flavanonol; a flavan; anthocyanin; isoflavane; new flavanthamine; isoflavone; procyanidins; phenolic acid; hydroxycinnamic acid; coumarin; stilbenes; anthraquinone; lignans; lignin; tannin; a polyphenol protein; catechin; rutin; robinia pseudoacacia; genistein; kaempferol; gallocatechin; catechin gallate; epicatechin; epigallocatechin; epicatechin gallate; quercetin; catechin; gallocatechin gallate; epicatechin; epigallocatechin; epicatechin gallate; epigallocatechin gallate; kaempferol; quercetin; naringin; neohesperidin; eriocitrin; naringin; naringenin; hesperidin; rhus verniciflua Stokes glycoside; myrosin; melissa glucoside; hesperetin; poncirin; epicatechin; gallocatechin; epigallocatechin; coumaric acid; cinnamic acid; gallic acid; ellagic acid; protocatechuic acid; chlorogenic acid; caffeic acid; ferulic acid; punicalagin; pomegranate rind tannagin.

H. A method according to paragraph B, wherein the polyphenol-enriched plant is selected from the group consisting of: allium species; a Brassica (Brassica) species; a camellia (Camelia) species; a Capsicum (Capsicum) species; citrus (Citrus) species; citrus aurantium (Citrus aurantium); cucumber (Cucumis) species; malus (Malus) species; a Musa (Musa) species; a Phaseolus (Phaseolus) species; a prune (prune) species; a Punica (Punica) species; pear (Pyrus) species; solanum (Solanum) species and bilberry (vaccinum) species.

I. A method of reducing nitrogen excretion by a ruminant and/or reducing methane emissions by a ruminant and/or increasing nitrogen-rich and carbon-rich material in a ruminant, the method comprising progressively administering to the ruminant an effective amount of at least one type of methanogenesis inhibitor.

J. A method according to paragraph I, wherein the step-wise administration of the methanogenesis inhibitor is performed with at least one dose, the percentage of which based on the weight of the ruminant being selected from the group consisting of: 0.1%;0.2%;0.3%;0.4%;0.5%;0.6%;0.7%;0.8%;0.9%;1.0%;1.1%;1.2%;1.3%;1.4%;1.5%;2.0%;2.5%;3.0%;3.5%;4.0%;4.5%;5.0%; and 10%.

K. A method according to paragraph I, wherein the step-wise administration of the methanogenesis inhibitor is performed with at least one dose, the percentage of which in the feed weight of the ruminant is selected from the group consisting of: 0.01%,0.03%,0.05%, 0.075%, 0.1%;0.2%;0.3%;0.4%;0.5%;0.6%;0.7%;0.8%;0.9%;1.0%;1.1%;1.2%;1.3%;1.4%;1.5%;2.0%;2.5%;3.0%;3.5%;4.0%;4.5%;5.0%; and 10%.

L. the method according to any one of paragraphs I to K, wherein stepwise administration has at least one interval between successive doses, said interval being selected from the group consisting of: 1 minute; 1 hour; 1 day; for 2 days; 3 days; 4 days; for 5 days; for 6 days; 7 days; for 10 days; 2 weeks; 3 weeks; 4 weeks; for 6 weeks; 2 months; 3 months; 4 months; 6 months; 9 months; and 12 months.

Claims

1. A composition for reducing methane emissions comprising an organohalogen compound and an organosulfur compound.

2. The composition of any preceding claim, wherein the organohalogen compound is an organobromine compound.

3. The composition of any of the preceding claims, wherein the organohalogen is bromoform.

4. The composition of any one of the preceding claims, wherein the organosulfur compound is from a plant of the Allium (Allium) species.

5. The composition of any of the preceding claims, wherein the organosulfur compound is allicin (C ₆ H ₁₀ S ₂ O); diallyl sulfide (C) ₆ H ₁₀ S) S; diallyl disulfide (C) ₆ H ₁₀ S ₂ ) The method comprises the steps of carrying out a first treatment on the surface of the And allyl mercaptan (C) ₃ H ₆ S)。

6. The composition of any of the preceding claims, wherein the organosulfur compound is allicin (C ₆ H ₁₀ S ₂ O)。

7. The composition of any of the preceding claims, wherein the ratio of organohalogen to organosulfur is from 1:10 to 1:3500.

8. The composition of any of the preceding claims, wherein the ratio of organohalogen to organosulfur is from 1:1000 to 1:2500.

9. The composition of any preceding claim, further comprising at least one polyphenol compound.

10. The composition of claim 9, wherein the at least one polyphenolic compound comprises at least one bioflavonoid.

11. The composition of claim 9, wherein the at least one polyphenolic compound comprises naringin, neohesperidin, or a combination thereof.

12. The composition of any preceding claim for use in inhibiting one or more methanogens.

13. The composition of any preceding claim for use in improving metabolic efficiency in an animal.

14. An animal feed comprising the composition of any one of claims 1-13.

15. Use of the composition according to claims 1-13 or the animal feed according to claim 14 for reducing methane emission and/or inhibiting one or more methanogens and/or improving the metabolic efficiency of an animal.

16. A method of reducing methane emissions, the method comprising administering to an animal the composition of any one of claims 1-13 or the animal feed of claim 14.

17. A method of inhibiting one or more methanogens, the method comprising administering to an animal the composition of any one of claims 1-13 or the animal feed of claim 14.

18. A method of improving the metabolic efficiency of an animal, the method comprising administering to the animal the composition of any one of claims 1-13 or the animal feed of claim 14.

19. The method of claims 16-18, wherein the animal is a ruminant, preferably wherein the ruminant is a cow, a goat or a sheep.