Compositions for reducing methane emission, animal feed comprising their and their use for reducing methane emission

Abstract

Compositions, such as animal feed compositions, for reducing methane emissions and / or for inhibiting one or more methanogens (i.e. Methanogens) are described. The use of the compositions for reducing methane emissions from an animal, such as a ruminant, for inhibiting methane production by one or more methanogens in the animal, and / or for increasing milk yield and / or weight gain in the animal, and related methods are described.

Description

Field of the Invention

[0001] The present invention relates to compositions for reducing methane emissions and / or for inhibiting one or more methanogenic organisms (i.e., methanogens), such as animal feed compositions. The present invention also relates to the use of said compositions for reducing methane emissions from animals such as ruminants, for inhibiting methane production by one or more methanogens in said animals, and / or for increasing milk production and / or weight gain of said animals, and related methods. Background of the Invention

[0003] Ruminants consume feed containing carbon and nitrogen and convert it into a variety of substances including carbon-rich lipids, fats, fatty acids and carbohydrates and nitrogen-rich proteins, nucleic acids, amino acids and nucleotides.

[0004] However, ruminants, such as cattle, sheep and goats, are inefficient users of carbon. The production of methane from carbon by ruminants represents an energy loss, accounting for 2% to 12% of the total energy intake from feed. Specific types of microorganisms in the ruminant gut use the end products of fermentation of nutrients eaten by the animal to produce methane as a byproduct. In addition to the negative impact of methane on climate change, this also reduces the nutrients available to the animal, for example, nutrients available to the animal for increased milk production and weight gain.

[0005] There is a need to develop alternative and / or improved compositions for reducing methane emissions from animals, such as ruminants, for example by increasing the availability of nutrients to the animals, and / or for increasing milk production and / or weight gain in such animals.

[0006] The present invention addresses these needs by providing compositions, such as compositions in the form of animal feeds (e.g., animal feed supplements, animal feed additives and complementary feeds), which reduce methane emissions into the environment and / or which increase milk production and / or weight gain in animals and more generally improve animal growth.

[0007] The present invention satisfies this need, at least in part, by providing compositions that inhibit methanogenesis by methanogens. SUMMARY OF THE INVENTION

[0009] According to the present invention, a first aspect provides a composition for reducing methane emissions, which comprises trihalomethane other than dibromochloromethane and at least one other compound selected from the group consisting of dibromochloromethane, dibromomethane (DBM), diallyl disulfide (DADS), diallyl trisulfide (DATS), dimethyl disulfide, dipropyl disulfide (DPDS), isoprenylnaringenin (e.g., 6-isoprenylnaringenin or 8-isoprenylnaringenin), bavachin, bavachinin, corylifol (e.g., corylifol A or corylifol C), sangsin , isobavachalcone, naringenin, quercetin, xanthohumol, tannic acid, cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil, lemon verbena oil, carvacrol, citral, cinnamaldehyde, hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), sclareol, saikosaponins (e.g., saikosaponin A, saikosaponin b2, saikosaponin C or saikosaponin D), ursolic acid, ginsenosides (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1 or ginsenoside Rf), dibromoacetic acid, 3-nitrooxypropanol, pyromellitic acid diimide, and any combination thereof. In certain embodiments, the composition may consist essentially of trihalomethanes other than dibromochloromethane and the compound(s), or consist of trihalomethanes other than dibromochloromethane and the compound(s). The composition is suitable for reducing methane emissions from animals, and / or inhibiting methane production by one or more methanogens in animals, and / or increasing milk production and / or weight gain in animals, e.g., ruminants such as cattle, goats, or sheep.

[0010] Furthermore, according to the present invention, a second aspect provides an animal feed, such as an animal feed supplement, an animal feed additive or a supplementary feed, comprising the composition of the first aspect or any embodiment thereof.

[0011] In addition, according to the present invention, the third aspect provides the use of the composition of the first aspect or the animal feed of the second aspect for reducing methane emission or production in animals, and / or inhibiting methane production by one or more methanogens in animals, and / or increasing milk production and / or weight gain in animals, for example ruminants such as cattle, goats or sheep.

[0012] In addition, according to the present invention, a fourth aspect provides a method for reducing methane emissions from animals, and / or inhibiting methane production by one or more methanogens in animals, and / or increasing milk production and / or weight gain in animals, the method comprising administering the composition to the animal, more specifically to a ruminant such as a cattle, goat or sheep (for example, by the animal consuming the composition or animal feed).

[0013] Embodiments of the invention are further described in the detailed description.Any embodiment described herein or any combination of embodiments described herein is applicable to any one or more aspects of the invention unless clearly contradicted by the context. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 : Methane inhibition by halomethane analogs as single compounds.

[0015] Figure 2 : Methane inhibition by iodoform and dibromomethane (DBM) as single compounds and in combination.

[0016] Figure 3 : Methane inhibition by iodoform and pyromellitic diimide (PD) as single compounds and in combination.

[0017] Figure 4 : Methane inhibition by iodoform and naringenin as single compounds and in combination.

[0018] Figure 5 : Methane inhibition by iodoform and dipropyl disulfide (DPDS) as single compounds and in combination.

[0019] Figure 6 : Methane inhibition by iodoform and garlic oil as single compounds and in combination.

[0020] Figure 7 : Methane inhibition by iodoform and diallyl trisulfide (DATS) as single compounds and in combination.

[0021] Figure 8 : Methane inhibition by iodoform and diallyl disulfide (DADS) as single compounds and in combination.

[0022] Fig. 9 : Methane inhibition by iodoform and α-hexylcinnamaldehyde as single compounds and in combination.

[0023] Fig.10 : Methane inhibition by iodoform and thyme red oil (TRO) as single compounds and in combination.

[0024] Fig.11: Methane inhibition by iodoform and lemongrass oil (LGO) as single compounds and in combination.

[0025] Fig.12 : Methane inhibition by iodoform, dibromomethane (DBM), and naringenin as single compounds and in combination.

[0026] Fig.13 : Methane inhibition by iodoform, dibromomethane (DBM) and diallyl trisulfide (DATS) as single compounds and in combination.

[0027] Fig.14 : Methane inhibition by iodoform, dibromomethane, and garlic oil as single compounds and in combination.

[0028] Fig.15 : Methane inhibition by iodoform, dibromomethane and diallyl disulfide (DADS) as single compounds and in combination.

[0029] Fig.16 : Methane inhibition by iodoform, dibromomethane and dipropyl disulfide (DPDS) as single compounds and in combination. DETAILED DESCRIPTION OF THE INVENTION

[0031] The present invention is based, at least in part, on the surprising discovery that a combination of a trihalomethane other than dibromochloromethane, such as bromoform (CHBr3) or iodoform (CHI3), and at least one other compound described herein, can reduce methane production and / or emissions in animals, such as ruminants. Without intending to be bound by theory, it is believed that a trihalomethane other than dibromochloromethane, such as bromoform (CHBr3) or iodoform (CHI3), together with at least one other compound, acts synergistically to inhibit methane production by methanogens, such as methanogens in the rumen and hindgut of animals, such as ruminants, thereby inhibiting methane production. "Acting synergistically together" means that the methane production inhibition capacity of a composition containing a mixture of a trihalomethane other than dibromochloromethane, such as bromoform (CHBr3) or iodoform (CHI3), and at least one other compound described herein is greater than the sum of the methane production inhibition capacities of either component alone. Inhibition of methanogenesis is believed to occur through different modes of action including, for example: reducing the methanogenic process; reducing methanogens by limiting, inhibiting or preventing enzymes involved in methanogenesis from participating in the process; or reducing methanogens by limiting their growth or killing them.

[0032] The present invention will be described below according to the preferred embodiments of the present invention and with reference to the accompanying specification. However, it should be understood that the description is limited to the preferred embodiments of the present invention only for the convenience of discussion of the present invention, and it is envisioned that those skilled in the art can design various modifications without departing from the scope of the appended claims.

[0033] When the term "at least one" or "one or more" appears above and below, this particularly means one to twenty or one to ten, preferably one to five or one to three, and particularly means one or further two of the listed features such as components. When ranges are indicated, such as weight percentage ranges, these ranges include the indicated limit values; thus, for example, "X to Y" means "from X and including X up to Y and including Y".

[0034] The term "therapeutic treatment" or "method of treatment" also includes prevention and alleviation of symptoms of a disease and / or disorder in a subject, but does not include cosmetic treatments. As used herein, the expression "treat or prevent" and similar terms refer to all forms of medical care, both preventive and curative, intended to eliminate or avoid a disease and / or disorder or to alleviate the symptoms thereof, as judged by any test available in current medical practice. Interventions that are intended with a reasonable expectation to achieve a specific result, but not always so, are included in the expression "treat or prevent". Interventions that successfully slow or stop the progression of a disease and / or disorder are included in the expression "treat or prevent".

[0035] The term "organohalogen" refers to an organic compound containing at least one halogen.

[0036] The term "trihalomethane" refers to a CHX3 substance, wherein X is a halogen, such as Cl, Br, I or a mixture thereof. X is preferably Br or I. Trihalomethanes are different from dibromochloromethane. Advantageously, trihalomethanes are bromoform (CHBr3) and / or iodoform (CHI3).

[0037] The term "consisting of does not include any additional elements, steps or ingredients not expressly recited.

[0038] The term "consisting essentially of " does not include any other key element, step or composition that is not clearly recorded, unless the other key element, step or composition does not have a substantial impact on the basic and novel properties of the present invention. When one or more other (multiple) key elements, (multiple) steps or (multiple) ingredients are one or more other (multiple) components of a product or composition, the total amount of other (multiple) components in the composition can be, for example, limited to 20 weight %. For example, the total amount of other (multiple) components in the composition can be limited to 19 weight % or 18 weight % or 17 weight % or 16 weight % or 15 weight % or 14 weight % or 13 weight % or 12 weight % or 11 weight % or 10 weight % or 9 weight % or 8 weight % or 7 weight % or 6 weight % or 5 weight % or 4 weight % or 3 weight % or 2 weight % or 1 weight %.

[0039] According to the present invention, there is provided a composition for reducing methane emission / production in animals, for example, comprising a trihalomethane (e.g., bromoform or iodoform) other than dibromochloromethane and at least one other compound selected from the group consisting of dibromochloromethane, dibromomethane, diallyl disulfide, diallyl trisulfide, dimethyl disulfide, dipropyl disulfide, isoprenylnaringenin (e.g., 6-isoprenylnaringenin or 8-isoprenylnaringenin), psoralen dihydroflavone, psoralen dihydroflavone methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sansin, isopsoralen chalcone, Naringenin, quercetin, xanthohumol, tannic acid, cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil, lemon verbena oil, carvacrol, citral, cinnamaldehyde, hexylcinnamaldehyde (e.g., α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde), sclareol, saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C, or saikosaponin D), ursolic acid, ginsenoside (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, or ginsenoside Rf), dibromoacetic acid, 3-nitrooxypropanol, pyromellitic acid diimide, and any combination thereof. The composition can be used to inhibit the production of methane by one or more methanogens, or to increase milk production and / or weight gain in animals.

[0040] Alternatively, the at least one other compound can be selected from dibromochloromethane, dibromomethane, diallyl disulfide, diallyl trisulfide, dimethyl disulfide, dipropyl disulfide, 8-isoprenylnaringenin, psoralen dihydroflavone, psoralen dihydroflavone methyl ether, chalcone A, sansin, isopsoralen chalcone, naringenin, quercetin, silymarin, xanthohumol, tannic acid, cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil, lemon verbena oil, carvacrol, citral, cinnamaldehyde, α-hexylcinnamaldehyde, rosin acid, sclareol, tanshinone IIA, bupleurosaponin b2, ursolic acid, ginsenoside Rb2, dibromoacetic acid, 3-nitrooxypropanol, pyromellitic acid diimide and any combination thereof.

[0041] Trihalomethanes are different from dibromochloromethane. Trihalomethanes can be compounds in which the halogens are of the same type. Trihalomethanes can be bromoform (CHBr3) and / or iodoform (CHI3). Trihalomethanes can be bromoform (CHBr3). Trihalomethanes can be iodoform (CHI3).

[0042] For example, a composition for reducing methane emissions / production in animals can include bromoform and at least one other compound selected from the group consisting of dibromochloromethane, diallyl disulfide, diallyl trisulfide, dimethyl disulfide, dipropyl disulfide, isoprenylnaringenin (e.g., 6-isoprenylnaringenin or 8-isoprenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sangsin, isopsoralen chalcone, naringenin, quercetin, xanthohumol, tannic acid, cinnamic acid, chalcones ... Peel oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil, lemon verbena oil, carvacrol, citral, cinnamaldehyde, hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), sclareol, saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C or saikosaponin D), ursolic acid, ginsenosides (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1 or ginsenoside Rf), dibromoacetic acid, 3-nitrooxypropanol and any combination thereof.

[0043] For example, a composition for reducing methane emissions / production in animals may comprise iodoform and at least one other compound selected from the group consisting of dibromomethane, pyromellitic acid diimide, naringenin, dipropyl disulfide, garlic oil, diallyl disulfide, diallyl trisulfide, hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde), thyme red oil, lemongrass oil, and any combination thereof.

[0044] Trihalomethanes can be obtained by synthesis or from natural sources. For example, a biological source of an organohalogen compound such as bromoform or iodoform is a marine macroalgae rich in organohalogens. The marine macroalgae rich in organohalogens include at least one marine macroalgae selected from the group consisting of Asparagopsis armata; Asparagopsis taxiformis; Dictyota species; Oedogonium species; Ulva species; and Cladophora patentiramea. Thus, in some embodiments, the trihalomethane compounds are derived from marine microalgae rich in organohalogens, such as marine microalgae selected from the group consisting of Asparagopsis armata; Asparagopsis taxiformis; Dictyota species; Oedogonium species; Ulva species; and Cladophora patentiramea.

[0045] The trihalomethane compounds can be produced by bacteria, fungi and cyanobacteria. For example, the bacteria include one selected from the group consisting of Streptomyces sp. and Zobellia galactanivorans. For example, the fungus comprises a fungus selected from the group consisting of Pyricularia oryzae, Curvularia inaequalis, Pyrenophora tritici-repentis, Embellisia didymospora, Agaricus species, such as Agaricus angusticystidiatus, Agaricus iodoformicus Speg., Agaricus moelleri, Agaricus phaeolepidotus and Agaricus pseudochitonia, Mycena species, such as Mycena pipterygia (Scop.: Fr.) Gray and Mycena arcangeliana, Jahnoporus species, such as Jahnoporus pubescens. hirtus), Cortinarius species, such as Cortinarius obtusus (Fr.) and Cortinarius seidlii and Hemileccinum impolitum. For example, the cyanobacteria include a cyanobacteria selected from the group consisting of Trichodesmium erythraeum, Synechococcus sp., and Acaryochloris marina.

[0046] The trihalomethanes may be synthetic, ie, the trihalomethanes are chemically synthesized.

[0047] Trihalomethanes can be produced by recombinant yeast.

[0048] Bromoform can be obtained by synthesis or from natural sources. For example, the biological source of organohalogen compounds such as bromoform is marine macroalgae rich in organohalogens. Marine macroalgae species include Asparagopsis armata; Asparagopsis taxiformis; Dictyota species; Oedogonium species; Ulva species; and Cladophora patentiramea. Bromoform can be produced by bacteria, fungi and cyanobacteria. For example, the bacteria include Streptomyces spp. and Zobellia galactanivorans. For example, the fungi include Pyricularia oryzae, Curvularia inaequalis, Pyrenophoratritici-repentis and Embellisia didymospora. For example, the cyanobacteria include Trichodesmium erythraeum, Synechococcus sp., and Acaryochloris marina.

[0049] In other embodiments, the bromoform is synthetic, ie, chemically synthesized.

[0050] In other embodiments, bromoform can be produced by recombinant yeast.

[0051] Iodoform can be obtained by synthesis or from natural sources. For example, the biological source of organohalogen compounds such as bromoform is marine macroalgae rich in organohalogens. Marine macroalgae species include Asparagopsis armata; Asparagopsis taxiformis; Dictyota species; Oedogonium species; Ulva species; and Cladophora patentiramea. Iodoform can be produced by bacteria, fungi and cyanobacteria. For example, the bacteria include Streptomyces sp. and Zobellia galactanivorans. For example, the fungus comprises a fungus selected from the group consisting of Pyricularia oryzae, Curvularia inaequalis, Pyrenophora tritici-repentis, Embellisia didymospora, Agaricus species, such as Agaricus angusticystidiatus, Agaricusiodoformicus Speg., Agaricus moelleri, Agaricus phaeolepidotus and Agaricus pseudochitonia, Mycena species, such as Mycena pipterygia (Scop.: Fr.) Gray and Mycena arcangeliana, Jahnoporus species, such as Jahnoporus pubescens. hirtus, Cortinarius species, such as Cortinarius obtusus (Fr.) and Cortinarius seidlii and Hemileccinum impolitum. For example, the cyanobacteria include Trichodesmium erythraeum, Synechococcus sp. and Acaryochlorismarina.

[0052] In other embodiments, the iodoform is synthetic, ie, chemically synthesized.

[0053] In other embodiments, iodoform can be produced by recombinant yeast.

[0054] In some embodiments, the at least one other compound is dibromochloromethane. The compound can be characterized as an organohalogen. It can be derived from a natural product or synthesized.

[0055] In some embodiments, the at least one other compound is dibromomethane. The compound can be characterized as an organohalogen. It can be derived from a natural product or synthesized.

[0056] In some embodiments, the at least one other compound is selected from diallyl disulfide, diallyl trisulfide, dimethyl disulfide and dipropyl disulfide. These compounds may be referred to as organosulfur compounds. They may be derived from natural products or synthesized. For example, organosulfur compounds may be obtained from garlic or another Allium species. For example, organosulfur compounds may be obtained from extracts of Allium species such as garlic (Allium sativum).

[0057] In some embodiments, the at least one other compound is selected from prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sangsin, isopsoralen chalcone, naringenin, quercetin, and xanthohumol. These can be characterized as flavonoid / flavonoid compounds. They can be derived from natural products or synthesized. In one embodiment, the prenylnaringenin is 8-prenylnaringenin. In another embodiment, the chalcone glycosides is chalcone glycosides A.

[0058] In some embodiments, the at least one other compound is tannic acid. The compound can be characterized as a polyphenol. It can be derived from a natural product or synthesized.

[0059] In some embodiments, the at least one other compound is selected from cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil and lemon verbena oil. These can be characterized as essential oils. They can be derived from natural products or synthesized.

[0060] In some embodiments, the at least one other compound is selected from carvacrol, citral, cinnamaldehyde and hexylcinnamaldehyde (e.g., α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde). These can be characterized as components of essential oils. They can be derived from natural products, in particular from essential oils, or synthesized. In one embodiment, the hexylcinnamaldehyde is α-hexylcinnamaldehyde.

[0061] In some embodiments, the at least one other compound is selected from sclareol, saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C, or saikosaponin D), ursolic acid, and ginsenosides (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, or ginsenoside Rf). These can be characterized as terpenes. These can be derived from natural products or synthesized. In one embodiment, saikosaponin is saikosaponin b2. In another embodiment, ginsenoside is ginsenoside Rb2.

[0062] In some embodiments, the at least one other compound is dibromoacetic acid. This can be characterized as a carboxylic acid. It can be derived from a natural product or synthesized.

[0063] In some embodiments, the at least one other compound is 3-nitrooxypropanol. This can be characterized as a nitrooxy compound.

[0064] In some embodiments, the at least one other compound is a pyromellitic acid imide. This can be characterized as an arylene imide.

[0065] - Ratio of trihalomethanes to at least one other compound(s)

[0066] In some embodiments, the concentration ratio of trihalomethanes other than dibromochloromethane to at least one other compound is from about 1:0.05 to about 1:10000. For example, in some embodiments, the concentration ratio of trihalomethanes to at least one other compound is from about 1:0.05 to about 1:100, or from about 1:0.1 to about 1:75, or from about 1:0.1 to about 1:50. In some embodiments, the ratio of trihalomethane to at least one other compound is from about 1:50 to about 1:9000, or from about 1:100 to about 1:9000, or from about 1:500 to about 1:9000, or from about 1:50 to about 1:100, or from about 1:100 to about 1:1000, or from about 1:100 to about 1:500, or from about 1:500 to about 1:1500, or from about 1:1000 to about 1:3000, or from about 1:10 to about 1:100, or from about 1:1000 to about 1:3000, or from about 1:3000 to about 1:5000, or from about 1:7000 to about 1:9000, or from about 1:8000 to about 1:8500.

[0067] In some embodiments, the at least one other compound is dibromochloromethane, and the concentration ratio of trihalomethanes other than dibromochloromethane to dibromochloromethane is from about 1:1 to about 1:10, or from about 1:1 to about 1:5, or from about 1:1 to about 1:2, or about 1:1, or about 1:1.2, or about 1:1.3, or about 1:1.4, or about 1:1.5, or about 1:1.6, or about 1:1.7, or about 1:1.8, or about 1:1.9.

[0068] In some embodiments, the at least one other compound is dibromomethane, and the concentration ratio of trihalomethanes other than dibromochloromethane to dibromomethane is from about 1:0.05 to about 1:1, or from about 1:0.1 to about 1:0.75, or from about 1:0.15 to about 1:0.5, or from about 1:0.2 to about 1:0.3.

[0069] In some embodiments, the at least one other compound is selected from diallyl disulfide, diallyl trisulfide, dimethyl disulfide and dipropyl disulfide, and the concentration ratio of trihalomethane other than dibromochloromethane to the compound is about 1:100 to about 1:10000, such as about 1:500 to about 1:10000, or about 1:750 to about 1:1250, or about 1:7500 to about 1:9000. In some embodiments, the at least one other compound is selected from diallyl disulfide, diallyl trisulfide, dimethyl disulfide and dipropyl disulfide, and the concentration ratio of trihalomethane to the compound is about 1:1 to about 1:100, or about 1:5 to about 1:50, or about 1:10 to about 1:25, or about 1:10 to about 1:20.

[0070] In some embodiments, the at least one other compound is diallyl disulfide, and the ratio of trihalomethane other than dibromochloromethane to the diallyl disulfide is from about 1:100 to about 1:3000, for example, from about 1:100 to about 1:2500, or from about 1:100 to about 1:2100. In some embodiments, the at least one other compound is diallyl disulfide, and the ratio of trihalomethane to the diallyl disulfide is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0071] In some embodiments, the at least one other compound is diallyl trisulfide, and the concentration ratio of trihalomethane other than dibromochloromethane to the diallyl trisulfide is from about 1:500 to about 1:1500, such as from about 1:750 to about 1:1250, or from about 1:1000 to about 1:1100. In some embodiments, the at least one other compound is diallyl trisulfide, and the concentration ratio of trihalomethane to the diallyl trisulfide is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0072] In some embodiments, the at least one other compound is dimethyl disulfide, and the concentration ratio of trihalomethanes other than dibromochloromethane to dimethyl disulfide is about 1:7500 to 1:9000, such as about 1:8000 to 1:8500.

[0073] In some embodiments, the at least one other compound is dipropyl disulfide, and the concentration ratio of trihalomethanes other than dibromochloromethane to dipropyl disulfide is about 1:7500 to about 1:9000, for example, about 1:8000 to about 1:8500. In some embodiments, the at least one other compound is dipropyl disulfide, and the concentration ratio of trihalomethanes to dipropyl disulfide is about 1:1 to about 1:100, or about 1:5 to about 1:50, or about 1:10 to about 1:25, or about 1:12.5 to about 1:20, or about 1:12.5 to about 1:17.5.

[0074] In some embodiments, the at least one other compound is selected from prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sangsin, isopsoralen chalcone, naringenin, quercetin, and xanthohumol, and the concentration ratio of trihalomethane other than dibromochloromethane to the compound is about 1:10 to about 1:1000, such as about 1:25 to about 1:1000, or about 1:50 to about 1:100, or about 1:50 to about 1:150. In some embodiments, the at least one additional compound is selected from prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sangsin, isopsoralen chalcone, naringenin, quercetin, and xanthohumol, and the concentration ratio of trihalomethane to the compound is about 1:1 to about 1:500, or about 1:5 to about 1:250, or about 1:10 to about 1:100, or about 1:25 to about 1:75.

[0075] In some embodiments, the at least one other compound is prenylnaringenin, and the concentration ratio of trihalomethane other than dibromochloromethane to the compound is about 1:50 to about 1:150, such as about 1:75 to about 1:125, or about 1:100 to about 1:110. In one embodiment, the prenylnaringenin is 6-prenylnaringenin or 8-prenylnaringenin, preferably 8-prenylnaringenin.

[0076] In some embodiments, the at least one other compound is psoralen and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is about 1:100 to 1:300, such as about 1:150 to about 1:250, or 1:190 to 1:210.

[0077] In some embodiments, the at least one other compound is psoralen, and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is about 1:50 to 1:1000, such as about 1:100 to about 1:850, or 1:50 to 1:150, or about 1:75 to about 1:125, or about 1:100 to about 1:110.

[0078] In some embodiments, the at least one other compound is chalcone A or chalcone C, and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is about 1:10 to about 1:50, such as about 1:15 to about 1:35, or about 1:20 to about 1:30. In one embodiment, the at least one other compound is chalcone A.

[0079] In some embodiments, the at least one other compound is thiamethoxam, and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is from about 1:2 to about 1:100, for example, from about 1:5 to about 1:90, or from about 1:10 to about 1:90, or from about 1:10 to about 1:30, or from about 1:15 to about 1:25.

[0080] In some embodiments, the at least one other compound is isopsoralea chalcone, and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is from about 1:10 to about 1:100, such as from about 1:15 to about 1:75, or from about 1:20 to about 1:60, or from about 1:40 to about 1:60, or from about 1:45 to about 1:55.

[0081] In some embodiments, the at least one other compound is naringenin, and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is about 1:500 to about 1:1500, such as about 1:650 to about 1:1000, or about 1:750 to about 1:900, or about 1:800 to about 1:850. In some embodiments, the at least one other compound is naringenin, and the concentration ratio of trihalomethanes to the compound is about 1:1 to about 1:500, or about 1:5 to about 1:250, or about 1:10 to about 1:100, or about 1:25 to about 1:75, or about 1:40 to about 1:60.

[0082] In some embodiments, the at least one other compound is quercetin, and the concentration ratio of trihalomethane other than dibromochloromethane to the compound is from about 1:100 to about 1:1500, such as from about 1:250 to about 1:1000, or from about 1:350 to about 1:900, or from about 1:750 to about 1:1000, or from about 1:800 to about 1:850.

[0083] In some embodiments, the at least one other compound is xanthohumol, and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is about 1:25 to about 1:150, such as about 1:40 to about 1:120, or about 1:80 to about 1:120, or about 1:100 to about 1:110.

[0084] In some embodiments, the at least one other compound is tannic acid, and the concentration ratio of trihalomethane other than dibromochloromethane to the compound is about 1:500 to about 1:1500, such as about 1:750 to about 1:1000, or about 1:800 to about 1:900, or about 1:800 to about 1:850.

[0085] In some embodiments, the at least one other compound is selected from cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil and lemon verbena oil, and the concentration ratio of trihalomethanes other than dibromochloromethane to the amount of the compound is about 1:100 to about 1:10000, such as about 1:200 to about 1:9000, or about 1:500 to about 1:2500, or about 1:3000 to about 1:5000, or about 1:7500 to 1:10000, or about 1:200 to about 1:300. In some embodiments, the at least one other compound is selected from cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil and lemon verbena oil, and the concentration ratio of trihalomethanes to the amount of the compound is about 1:0.5 to about 1:100, or about 1:1 to about 1:75, or about 1:2.5 to about 1:50, or about 1:5 to about 1:30.

[0086] In some embodiments, the at least one other compound is cassia oil, and the concentration ratio of trihalomethanes other than dibromochloromethane to the amount of the compound is from about 1:7500 to about 1:10000, or from about 1:8000 to about 1:9000, or from about 1:8000 to about 1:8500.

[0087] In some embodiments, the at least one other compound is garlic oil, and the concentration ratio of trihalomethanes other than dibromochloromethane to the amount of the compound is about 1:500 to about 1:4000, or about 1:750 to about 1:3000, or about 1:1000 to about 1:2500, or about 1:1500 to about 1:2500, or about 1:1750 to about 1:2250, or about 1:2000 to about 1:2100. In some embodiments, the at least one other compound is garlic oil, and the concentration ratio of trihalomethanes to the amount of the compound is about 1:0.5 to about 1:100, about 1:1 to about 1:50, or about 1:2.5 to about 1:25, or about 1:5 to about 1:10.

[0088] In some embodiments, the at least one other compound is sandalwood oil, and the concentration ratio of the trihalomethane other than dibromochloromethane to the amount of the compound is from about 1:100 to about 1:500, or from about 1:200 to about 1:400, or from about 1:200 to about 1:300, or from about 1:250 to about 1:270.

[0089] In some embodiments, the at least one other compound is thyme red oil, and the concentration ratio of trihalomethanes other than dibromochloromethane to the amount of the compound is about 1:2500 to about 1:7500, or about 1:3000 to about 1:5000, or about 1:3750 to about 1:4500, or about 1:4000 to about 1:4500, or about 1:4100 to about 1:4250, or about 1:4100 to about 1:4200. In some embodiments, the at least one other compound is thyme red oil, and the concentration ratio of trihalomethanes to the amount of the compound is about 1:1 to about 1:100, about 1:2.5 to about 1:75, or about 1:5 to about 1:50, or about 1:10 to about 1:40, about 1:20 to about 1:30.

[0090] In some embodiments, the at least one other compound is lemongrass oil, and the concentration ratio of trihalomethanes other than dibromochloromethane to the amount of the compound is about 1:2500 to about 1:7500, or about 1:3000 to about 1:5000, or about 1:3750 to about 1:4500, or about 1:4000 to about 1:4500, or about 1:4100 to about 1:4250, or about 1:4100 to about 1:4200. In some embodiments, the at least one other compound is lemongrass oil, and the concentration ratio of trihalomethanes to the amount of the compound is about 1:1 to about 1:100, about 1:2.5 to about 1:75, or about 1:5 to about 1:50, or about 1:10 to about 1:40, about 1:20 to about 1:30.

[0091] In some embodiments, the at least one other compound is lemon verbena oil, and the concentration ratio of the trihalomethane other than dibromochloromethane to the amount of the compound is from about 1:2500 to about 1:7500, or from about 1:3000 to about 1:5000, or from about 1:3750 to about 1:4500, or from about 1:4000 to about 1:4500, or from about 1:4100 to about 1:4250, or from about 1:4100 to about 1:4200.

[0092] In some embodiments, the at least one other compound is selected from carvacrol, citral, cinnamaldehyde and hexylcinnamaldehyde (e.g., α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde), and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is from about 1:200 to about 1:10000, for example, from about 1:200 to about 1:300, or from about 1:3000 to about 1:9000, or from about 1:3000 to about 1:5000, or from about 1:7500 to about 1:10000. In some embodiments, the at least one other compound is selected from carvacrol, citral, cinnamaldehyde, and hexylcinnamaldehyde (e.g., α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde), and the concentration ratio of trihalomethane to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0093] In some embodiments, the at least one other compound is carvacrol, and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is from about 1:2500 to about 1:7500, or from about 1:3000 to about 1:5000, or from about 1:3750 to about 1:4500, or from about 1:4000 to about 1:4500, or from about 1:4100 to about 1:4250, or from about 1:4100 to about 1:4200.

[0094] In some embodiments, the at least one other compound is citral, and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is from about 1:2500 to about 1:7500, or from about 1:3000 to about 1:5000, or from about 1:3750 to about 1:4500, or from about 1:4000 to about 1:4500, or from about 1:4100 to about 1:4250, or from about 1:4100 to about 1:4200.

[0095] In some embodiments, the at least one other compound is cinnamaldehyde, and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is from about 1:7500 to about 1:10000, or from about 1:8000 to about 1:9000, or from about 1:8000 to about 1:8500.

[0096] In some embodiments, the at least one other compound is hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is about 1:100 to about 1:500, or about 1:200 to about 1:400, or about 1:200 to about 1:300, or about 1:250 to about 1:270. In some embodiments, the at least one other compound is hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), and the concentration ratio of trihalomethanes to the compound is about 1:1 to about 1:100, or about 1:5 to about 1:50, or about 1:10 to about 1:25, or about 1:10 to about 1:20, or about 1:10 to about 1:15. In one embodiment, the at least one other compound is α-hexyl cinnamaldehyde.

[0097] In some embodiments, the at least one other compound is selected from sclareol, saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C, or saikosaponin D), ursolic acid, and ginsenoside (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, or ginsenoside Rf), and the concentration ratio of trihalomethanes other than dibromochloromethane to the compounds is about 1:50 to about 1:1000, or about 1:50 to about 1:250, or about 1:750 to about 1:900.

[0098] In some embodiments, the at least one other compound is sclareol, and the concentration ratio of trihalomethane other than dibromochloromethane to the compound is from about 1:50 to about 1:500, or from about 1:100 to about 1:350, or from about 1:150 to about 1:250, or from about 1:175 to about 1:225.

[0099] In some embodiments, the at least one other compound is a saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C, or saikosaponin D), and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is from about 1:25 to about 1:150, or from about 1:50 to about 1:100, or from about 1:70 to about 1:90, or from about 1:80 to about 1:85. In one embodiment, the at least one other compound is saikosaponin b2.

[0100] In some embodiments, the at least one other compound is ursolic acid, and the concentration ratio of the trihalomethane other than dibromochloromethane to the compound is about 1:500 to about 1:1500, such as about 1:750 to about 1:1000, or about 1:800 to about 1:900, or about 1:800 to about 1:850.

[0101] In some embodiments, the at least one other compound is a ginsenoside (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, or ginsenoside Rf), and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is from about 1:50 to about 1:350, or from about 1:50 to about 1:250, or from about 1:100 to about 1:200, or from about 1:150 to about 1:175. In one embodiment, the at least one other compound is ginsenoside Rb2.

[0102] In some embodiments, the at least one other compound is dibromoacetic acid, and the concentration ratio of trihalomethanes other than dibromochloromethane to dibromoacetic acid is about 1:1000 to about 1:2000, such as about 1:1500 to about 1:1750, or about 1:1600 to about 1:1700.

[0103] In some embodiments, the at least one other compound is 3-nitrooxypropanol, and the concentration ratio of trihalomethane other than dibromochloromethane to 3-nitrooxypropanol is from about 1:2 to about 1:20, e.g., from about 1.5 to about 1:15, or from about 1:5 to about 1:10, or from about 1:6 to about 1:8.

[0104] In some embodiments, the at least one other compound is pyromellitic acid imide, and the concentration ratio of trihalomethane other than dibromochloromethane to pyromellitic acid imide is from about 1:0.05 to about 1:25, or from about 1:0.1 to about 1:10, or from about 1:0.5 to about 1:5, or from about 1:1 to about 1:2.5.

[0105] In some embodiments, the composition comprises trihalomethanes other than dibromochloromethane and at least two other compounds, such as trihalomethanes and two other compounds. In some embodiments, the composition comprises trihalomethanes other than dibromochloromethane and at least two other compounds, and the at least two other compounds are selected from dibromomethane, pyromellitic acid diimide, naringenin, dipropyl disulfide, garlic oil, diallyl disulfide, diallyl trisulfide, hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde), thyme red oil, lemongrass oil, and any combination thereof. In some embodiments, the composition comprises trihalomethanes other than dibromochloromethane, dibromomethane, and one other compound, and the one other compound is selected from naringenin, dipropyl disulfide, garlic oil, diallyl disulfide, or diallyl trisulfide.

[0106] In some embodiments, the concentration ratio of trihalomethanes other than dibromochloromethane to the at least two other compounds is from about 1:0.05:1 to about 1:1:500. For example, in some embodiments, the concentration ratio of trihalomethanes to the at least two other compounds is from about 1:0.05:1 to about 1:0.75:250, or from about 1:0.15:2 to about 1:0.5:100, or from about 1:0.2:5 to about 1:0.3:50.

[0107] In some embodiments, one of the at least two other compounds is dibromomethane, and the concentration ratio of trihalomethanes other than dibromochloromethane to dibromomethane is from about 1:0.05 to about 1:1, or from about 1:0.1 to about 1:0.75, or from about 1:0.15 to about 1:0.5, or from about 1:0.2 to about 1:0.3.

[0108] In some embodiments, one of the at least two other compounds is selected from diallyl disulfide, diallyl trisulfide, dimethyl disulfide, and dipropyl disulfide, and the concentration ratio of trihalomethane other than dibromochloromethane to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20.

[0109] In some embodiments, one of the at least two other compounds is diallyl disulfide, and the ratio of trihalomethane other than dibromochloromethane to the diallyl disulfide is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0110] In some embodiments, one of the at least two other compounds is diallyl trisulfide, and the concentration ratio of the trihalomethane other than dibromochloromethane to the diallyl trisulfide is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0111] In some embodiments, one of the at least two other compounds is dipropyl disulfide, and the concentration ratio of trihalomethanes other than dibromochloromethane to dipropyl disulfide is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:12.5 to about 1:20, or from about 1:12.5 to about 1:17.5.

[0112] In some embodiments, one of the at least two other compounds is selected from prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sangsin, isopsoralen chalcone, naringenin, quercetin, and xanthohumol, and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is about 1:1 to about 1:500, or about 1:5 to about 1:250, or about 1:10 to about 1:100, or about 1:25 to about 1:75.

[0113] In some embodiments, one of the at least two other compounds is naringenin, and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is from about 1:1 to about 1:500, or from about 1:5 to about 1:250, or from about 1:10 to about 1:100, or from about 1:25 to about 1:75, or from about 1:40 to about 1:60.

[0114] In some embodiments, one of the at least two other compounds is selected from cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil and lemon verbena oil, and the concentration ratio of trihalomethane other than dibromochloromethane to the amount of the compound is from about 1:0.5 to about 1:100, or from about 1:1 to about 1:75, or from about 1:2.5 to about 1:50, or from about 1:5 to about 1:30.

[0115] In some embodiments, one of the at least two other compounds is garlic oil, and the concentration ratio of trihalomethanes other than dibromochloromethane to the amount of the compound is from about 1:0.5 to about 1:100, from about 1:1 to about 1:50, or from about 1:2.5 to about 1:25 or from about 1:5 to about 1:10.

[0116] In some embodiments, one of the at least two other compounds is thyme red oil, and the concentration ratio of trihalomethanes other than dibromochloromethane to the amount of the compound is from about 1:1 to about 1:100, from about 1:2.5 to about 1:75, or from about 1:5 to about 1:50, or from about 1:10 to about 1:40, from about 1:20 to about 1:30.

[0117] In some embodiments, one of the at least two other compounds is lemongrass oil, and the concentration ratio of the trihalomethane other than dibromochloromethane to the amount of the compound is from about 1:1 to about 1:100, from about 1:2.5 to about 1:75, or from about 1:5 to about 1:50, or from about 1:10 to about 1:40, or from about 1:20 to about 1:30.

[0118] In some embodiments, one of the at least two other compounds is selected from carvacrol, citral, cinnamaldehyde and hexylcinnamaldehyde (e.g., α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde), and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0119] In some embodiments, one of the at least two other compounds is hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), and the concentration ratio of trihalomethanes other than dibromochloromethane to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15. In one embodiment, the at least one other compound is α-hexyl cinnamaldehyde.

[0120] In some embodiments, one of the at least two other compounds is pyromellitic acid imide, and the concentration ratio of trihalomethane other than dibromochloromethane to pyromellitic acid imide is from about 1:0.05 to about 1:25, or from about 1:0.1 to about 1:10, or from about 1:0.5 to about 1:5, or from about 1:1 to about 1:2.5.

[0121] In some embodiments, the at least two other compounds are dibromomethane and diallyl disulfide, and the concentration of trihalomethanes other than dibromochloromethane to dibromomethane to diallyl disulfide is from about 1:0.05:1 to about 1:1:50, or from about 1:0.1:5 to about 1:0.75:25, or from about 1:0.15:10 to about 1:0.5:20, or from about 1:0.2:10 to about 1:0.3:15.

[0122] In some embodiments, the at least two other compounds are dibromomethane and diallyl trisulfide, and the concentration of trihalomethanes other than dibromochloromethane to dibromomethane to diallyl trisulfide is from about 1:0.05:1 to about 1:1:50, or from about 1:0.1:5 to about 1:0.75:25, or from about 1:0.15:10 to about 1:0.5:20, or from about 1:0.2:10 to about 1:0.3:15.

[0123] In some embodiments, the at least two other compounds are dibromomethane and dipropyl disulfide, and the concentration of trihalomethanes other than dibromochloromethane to dibromomethane and dipropyl disulfide is from about 1:0.05:1 to about 1:1:50, or from about 1:0.1:5 to about 1:0.75:25, or from about 1:0.15:10 to about 1:0.5:20, or from about 1:0.2:12.5 to about 1:0.3:17.5.

[0124] In some embodiments, the at least two other compounds are dibromomethane and naringenin, and the concentration of trihalomethanes other than dibromochloromethane to dibromomethane to naringenin is from about 1:0.05:5 to about 1:1:250, or from about 1:0.1:10 to about 1:0.75:100, or from about 1:0.15:25 to about 1:0.5:75, or from about 1:0.2:40 to about 1:0.3:60.

[0125] In some embodiments, the at least two other compounds are dibromomethane and garlic oil, and the concentration of iodoform to dibromomethane to garlic oil is about 1:0.05:0.5 to about 1:1:100, or about 1:0.1:1 to about 1:0.75:50, or about 1:0.15:2.5 to about 1:0.5:25, or about 1:0.2:5 to about 1:0.3:10.

[0126] In some embodiments, the concentration of trihalomethanes other than dibromochloromethane in the composition is less than 0.30 μM, such as less than 0.25 μM, or less than 0.20 μM, or less than 0.15 μM. In some embodiments, the concentration of trihalomethanes in the composition is at least about 0.05 μM, such as at least about 0.075 μM, or at least about 0.10 μM. In some embodiments, the concentration is from about 0.05 μM to about 0.20 μM, such as from about 0.075 μM to about 0.175 μM, or from about 0.10 μM to about 0.15 μM, or from about 0.11 μM to about 0.13 μM. In some embodiments, the concentration of trihalomethanes in the composition is less than 25 μM, or less than 20 μM, or less than 15 μM, or less than 10 μM, or less than 5 μM, or less than 2.5 μM. In some embodiments, the concentration of trihalomethanes other than dibromochloromethane in the composition is at least about 0.5 μM, such as at least about 1 μM, or at least about 2 μM, or at least about 5 μM, or at least about 10 μM. In some embodiments, the concentration is from about 0.5 μM to about 50 μM, or from about 1 μM to about 25 μM, or from about 1.5 μM to about 12.5 μM.

[0127] In some embodiments, the concentration of the at least one other compound is at least about 0.5 μM, at least about 2.0 μM, such as at least about 10.0 μM, or at least about 50.0 μM, or at least about 100.0 μM, or at least about 250.0 μM, or at least about 500.0 μM, or at least about 750.0 μM. In some embodiments, the concentration of the at least one other compound is less than about 5.0 mM, such as less than about 2.5 mM, or less than about 1.0 mM, or less than about 750.0 μM, or less than about 500.0 μM, or less than about 250.0 μM, or less than about 100.0 μM, or less than about 50 μM, or less than about 10 μM, or less than about 5 μM.

[0128] In some embodiments, the concentration of at least one other compound in the composition is about 0.15 μM to about 1.0 mM, for example, about 0.5 μM to about 1.0 mM, or about 2.0 μM to about 750.0 μM, or about 2.0 μM to about 500.0 μM, or about 2.0 μM to about 250.0 μM, or about 2.0 μM to about 150.0 μM, or about 2.0 μM to about 100.0 μM.

[0129] In some embodiments, the at least one other compound is dibromochloromethane, and the concentration of dibromochloromethane in the composition is about 0.15 μM to about 1.0 μM, such as about 0.15 μM to about 0.5 μM, or about 0.15 μM to about 0.25 μM.

[0130] In some embodiments, the at least one other compound is dibromomethane, and the concentration of dibromomethane in the composition is about 0.05 μM to about 50 μM, or about 0.1 μM to about 25 μM, or about 0.25 μM to about 12.5 μM.

[0131] In some embodiments, the at least one other compound is diallyl disulfide, and the concentration of diallyl disulfide in the composition is about 5.0 μM to about 500.0 μM, for example, about 10.0 μM to about 350.0 μM, or about 15.0 μM to about 250.0 μM. In some embodiments, the at least one other compound is diallyl disulfide, and the concentration of diallyl disulfide in the composition is about 0.1 μM to about 100.0 μM, or about 0.5 μM to about 75.0 μM, or about 1.0 μM to about 50.0 μM, or about 5.0 μM to about 40.0 μM, or about 15.0 μM to about 30.0 μM.

[0132] In some embodiments, the at least one other compound is diallyl trisulfide, and the concentration of diallyl trisulfide in the composition is about 50.0 μM to about 250.0 μM, such as about 75.0 μM to about 200.0 μM, or about 100.0 μM to about 150.0 μM. In some embodiments, the at least one other compound is diallyl trisulfide, and the concentration of diallyl trisulfide in the composition is about 0.1 μM to about 100.0 μM, or about 0.5 μM to about 75.0 μM, or about 1.0 μM to about 50.0 μM, or about 5.0 μM to about 40.0 μM, or about 15.0 μM to about 30.0 μM.

[0133] In some embodiments, the at least one other compound is dimethyl disulfide, and the concentration of dimethyl disulfide in the composition is about 500.0 μM to about 1.5 mM, such as about 750 μM to about 1.25 mM, or about 900.0 μM to about 1.1 mM.

[0134] In some embodiments, the at least one other compound is dipropyl disulfide, and the concentration of dipropyl disulfide in the composition is about 500.0 μM to about 1.5 mM, for example, about 750.0 μM to about 1.25 mM, or about 900.0 μM to about 1.1 mM. In some embodiments, the at least one other compound is dipropyl disulfide, and the concentration of dipropyl disulfide in the composition is about 0.5 μM to about 100.0 μM, or about 1.0 μM to about 80.0 μM, or about 5.0 μM to about 60.0 μM, or about 10.0 μM to about 50.0 μM, or about 15.0 μM to about 40.0 μM, or about 20.0 μM to about 40.0 μM.

[0135] In some embodiments, the at least one other compound is prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), and the concentration of prenylnaringenin in the composition is about 2.0 μM to about 50.0 μM, e.g., about 5.0 μM to about 25.0 μM, or about 10.0 μM to about 15.0 μM. In one embodiment, the prenylnaringenin is 8-prenylnaringenin.

[0136] In some embodiments, the at least one other compound is psoralen, and the concentration of psoralen in the composition is about 5.0 μM to about 75.0 μM, such as about 10.0 μM to about 50.0 μM, or about 15.0 μM to about 35.0 μM, or about 20.0 μM to about 30.0 μM.

[0137] In some embodiments, the at least one other compound is psoralen, and the concentration of psoralen in the composition is about 2.0 μM to about 200.0 μM, such as about 5.0 μM to about 150.0 μM, or about 10.0 μM to about 125.0 μM.

[0138] In some embodiments, the at least one other compound is chalcone glycoside A or chalcone glycoside C, and the concentration of chalcone glycoside A or chalcone glycoside C in the composition is about 0.5 μM to about 10.0 μM, such as about 1.0 μM to about 7.5 μM, or about 2.0 μM to about 5.0 μM. In one embodiment, the at least one other compound is chalcone glycoside A.

[0139] In some embodiments, the at least one other compound is sangsin, and the concentration of sangsin in the composition is about 0.25 μM to about 50.0 μM, such as about 0.50 μM to about 25.0 μM, or about 1.0 μM to about 15.0 μM.

[0140] In some embodiments, the at least one other compound is psoralea chalcone, and the concentration of psoralea chalcone in the composition is about 1.0 μM to about 15.0 μM, such as about 2.5 μM to about 12.5 μM, or about 4.0 μM to about 10.0 μM, or about 5.0 μM to about 7.5 μM.

[0141] In some embodiments, the at least one other compound is naringenin, and the concentration of naringenin in the composition is about 25.0 μM to about 250.0 μM, such as about 50.0 μM to about 150.0 μM, or about 75.0 μM to about 125.0 μM, or about 90.0 μM to about 110.0 μM.

[0142] In some embodiments, the at least one other compound is quercetin, and the concentration of quercetin in the composition is about 25.0 μM to about 250.0 μM, such as about 50.0 μM to about 150.0 μM, or about 75.0 μM to about 125.0 μM, or about 90.0 μM to about 110.0 μM.

[0143] In some embodiments, the at least one other compound is xanthohumol, and the concentration of xanthohumol in the composition is about 2.0 μM to about 50.0 μM, such as about 5.0 μM to about 25.0 μM, or about 10.0 μM to about 15.0 μM.

[0144] In some embodiments, the at least one other compound is tannic acid, and the concentration of tannic acid in the composition is about 25.0 μM to about 250.0 μM, such as about 50.0 μM to about 150.0 μM, or about 75.0 μM to about 125.0 μM, or about 90.0 μM to about 110.0 μM.

[0145] In some embodiments, the at least one other compound is carvacrol, and the concentration of carvacrol in the composition is about 250.0 μM to about 1.0 mM, such as about 250 μM to about 750.0 μM, or about 400.0 μM to about 600.0 μM, or about 450.0 μM to about 550.0 μM.

[0146] In some embodiments, the at least one other compound is citral, and the concentration of citral in the composition is about 250.0 μM to about 1.0 mM, such as about 250 μM to about 750.0 μM, or about 400.0 μM to about 600.0 μM, or about 450.0 μM to about 550.0 μM.

[0147] In some embodiments, the at least one other compound is cinnamaldehyde, and the concentration of cinnamaldehyde in the composition is about 500.0 μM to about 1.5 mM, such as about 750 μM to about 1.25 mM, or about 900.0 μM to about 1.1 mM.

[0148] In some embodiments, the at least one other compound is hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), and the concentration of hexyl cinnamaldehyde in the composition is about 10.0 μM to about 75.0 μM, such as about 15.0 μM to about 50.0 μM, or about 20.0 μM to about 40.0 μM, or about 25.0 μM to about 35.0 μM. In one embodiment, the at least one other compound is α-hexyl cinnamaldehyde.

[0149] In some embodiments, the at least one other compound is sclareol, and the concentration of sclareol in the composition is about 5.0 μM to about 75.0 μM, such as about 10.0 μM to about 50.0 μM, or about 15.0 μM to about 35.0 μM, or about 20.0 μM to about 30.0 μM.

[0150] In some embodiments, the at least one other compound is a saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C, or saikosaponin D), and the concentration of saikosaponin in the composition is about 2.0 μM to about 25.0 μM, such as about 5.0 μM to about 15.0 μM, or about 8.0 μM to about 12.0 μM. In one embodiment, the at least one other compound is saikosaponin b2.

[0151] In some embodiments, the at least one other compound is ursolic acid, and the concentration of ursolic acid in the composition is about 25.0 μM to about 250.0 μM, such as about 50.0 μM to about 150.0 μM, or about 75.0 μM to about 125.0 μM, or about 90.0 μM to about 110.0 μM.

[0152] In some embodiments, the at least one other compound is a ginsenoside (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, or ginsenoside Rf), and the concentration of ginsenosides in the composition is about 5.0 μM to about 50.0 μM, for example, about 10.0 μM to about 40.0 μM, or about 15.0 μM to about 30.0 μM, or about 15.0 μM to about 25.0 μM. In one embodiment, the at least one other compound is ginsenoside Rb2.

[0153] In some embodiments, the at least one other compound is dibromoacetic acid, and the concentration of dibromoacetic acid in the composition is about 50.0 μM to about 500.0 μM, such as about 100.0 μM to about 350.0 μM, or about 150.0 μM to about 250.0 μM, or about 175.0 μM to about 225.0 μM.

[0154] In some embodiments, the at least one other compound is 3-nitrooxypropanol, and the concentration of 3-nitrooxypropanol in the composition is about 0.25 μM to about 4.0 μM, for example, about 0.50 μM to about 2.5 μM, or about 0.50 μM to about 1.75 μM, or about 0.50 μM to about 1.5 μM, or about 0.50 μM to about 1.0 μM, or about 0.70 μM to about 0.85 μM.

[0155] In some embodiments, the at least one other compound is cassia oil, and the amount of cassia oil in the composition is from about 100 ppm to about 500 ppm, for example from about 150 ppm to about 350 ppm, or from about 200 ppm to about 300 ppm, or from about 225 ppm to about 275 ppm.

[0156] In some embodiments, the at least one other compound is garlic oil, and the amount of garlic oil in the composition is about 10 ppm to about 250 ppm, such as about 25 ppm to about 150 ppm, or about 40 ppm to about 100 ppm, or about 50 ppm to about 75 ppm. In some embodiments, the at least one other compound is garlic oil, and the amount of garlic oil in the composition is about 0.5 ppm, or about 1 ppm, or about 2 ppm, or about 3 ppm, or about 4 ppm, or about 5 ppm.

[0157] In some embodiments, the at least one other compound is sandalwood oil, and the amount of sandalwood oil in the composition is from about 1 ppm to about 20 ppm, for example from about 3 ppm to about 15 ppm, or from about 4 ppm to about 10 ppm, or from about 6 ppm to about 8 ppm, or from about 7 ppm to about 8 ppm.

[0158] In some embodiments, the at least one other compound is thyme red oil, and the amount of thyme red oil in the composition is from about 10ppm to about 250ppm, such as from about 50ppm to about 200ppm, or from about 75ppm to about 150ppm, or from about 100ppm to about 150ppm. In some embodiments, the at least one other compound is thyme red oil, and the amount of thyme red oil in the composition is from about 0.5ppm to about 50ppm, or from about 1ppm to about 40ppm, or from about 5ppm to about 35ppm, or from about 10ppm to about 30ppm, or from about 15ppm to about 25ppm.

[0159] In some embodiments, the at least one other compound is lemongrass oil, and the amount of lemongrass oil in the composition is from about 10 ppm to about 250 ppm, such as from about 50 ppm to about 200 ppm, or from about 75 ppm to about 150 ppm, or from about 100 ppm to about 150 ppm. In some embodiments, the at least one other compound is lemongrass oil, and the amount of lemongrass oil in the composition is from about 0.5 ppm to about 50 ppm, or from about 1 ppm to about 40 ppm, or from about 5 ppm to about 35 ppm, or from about 10 ppm to about 30 ppm, or from about 15 ppm to about 25 ppm.

[0160] In some embodiments, the at least one other compound is lemon verbena oil, and the amount of lemon verbena oil in the composition is from about 10 ppm to about 250 ppm, such as from about 50 ppm to about 200 ppm, or from about 75 ppm to about 150 ppm, or from about 100 ppm to about 150 ppm.

[0161] In some embodiments, the at least one other compound is pyromellitic acid imide, and the amount of pyromellitic acid imide in the composition is from about 0.1 ppm to about 10 ppm, or from about 0.25 ppm to about 7.5 ppm, or from about 0.5 to about 5 ppm, or from about 0.75 ppm to about 2.5 ppm.

[0162] In some embodiments, one of the at least two other compounds is dibromomethane, and the concentration of dibromomethane in the composition is about 0.05 μM to about 50 μM, or about 0.1 μM to about 25 μM, or about 0.25 μM to about 12.5 μM. In some embodiments, the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM.

[0163] In some embodiments, one of the at least two other compounds is diallyl disulfide, and the concentration of diallyl disulfide in the composition is from about 0.1 μM to about 100 μM, or from about 0.5 μM to about 75 μM, or from about 1.0 μM to about 50 μM, or from about 5.0 μM to about 40 μM, or from about 15 μM to about 30 μM.

[0164] In some embodiments, one of the at least two other compounds is diallyl trisulfide, and the concentration of diallyl trisulfide in the composition is from about 0.1 μM to about 100 μM, or from about 0.5 μM to about 75 μM, or from about 1.0 μM to about 50 μM, or from about 5.0 μM to about 40 μM, or from about 15 μM to about 30 μM.

[0165] In some embodiments, one of the at least two other compounds is dipropyl disulfide, and the concentration of dipropyl disulfide in the composition is about 0.5 μM to about 100 μM, or about 1.0 μM to about 80 μM, or about 5.0 μM to about 60 μM, or about 10 μM to about 50 μM, or about 15 μM to about 40 μM, or about 20 μM to about 40 μM.

[0166] In some embodiments, one of the at least two other compounds is naringenin, and the concentration of naringenin in the composition is about 25 μM to about 250 μM, such as about 50 μM to about 150 μM, or about 75 μM to about 125 μM, or about 90 μM to about 110 μM.

[0167] In some embodiments, one of the at least two other compounds is hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), and the concentration of hexyl cinnamaldehyde in the composition is about 10 μM to about 75 μM, such as about 15 μM to about 50 μM, or about 20 μM to about 40 μM, or about 25 μM to about 35 μM. In one embodiment, the at least one other compound is α-hexyl cinnamaldehyde.

[0168] In some embodiments, one of the at least two other compounds is garlic oil, and the amount of garlic oil in the composition is about 0.5 ppm, or about 1 ppm, or about 2 ppm, or about 3 ppm, or about 4 ppm, or about 5 ppm.

[0169] In some embodiments, one of the at least two other compounds is thyme red oil, and the amount of thyme red oil in the composition is from about 0.5 ppm to about 50 ppm, or from about 1 ppm to about 40 ppm, or from about 5 ppm to about 35 ppm, or from about 10 ppm to about 30 ppm, or from about 15 ppm to about 25 ppm.

[0170] In some embodiments, one of the at least two other compounds is lemongrass oil, and the amount of lemongrass oil in the composition is from about 0.5 ppm to about 50 ppm, or from about 1 ppm to about 40 ppm, or from about 5 ppm to about 35 ppm, or from about 10 ppm to about 30 ppm, or from about 15 ppm to about 25 ppm.

[0171] In some embodiments, one of the at least two other compounds is pyromellitic acid imide, and the amount of pyromellitic acid imide in the composition is from about 0.1 ppm to about 10 ppm, or from about 0.25 ppm to about 7.5 ppm, or from about 0.5 to about 5 ppm, or from about 0.75 ppm to about 2.5 ppm.

[0172] In some embodiments, the at least two other compounds are dibromomethane and diallyl disulfide, and the concentration of trihalomethanes other than dibromochloromethane in the composition is about 0.5 μM to about 10 μM, or about 1 μM to about 5 μM, or about 1.5 μM to about 2.5 μM; the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM; and the concentration of diallyl disulfide is about 1.0 μM to about 50 μM, or about 5.0 μM to about 40 μM, or about 15 μM to about 30 μM, or about 20 μM to about 30 μM.

[0173] In some embodiments, the at least two other compounds are dibromomethane and diallyl trisulfide, and the concentration of trihalomethanes other than dibromochloromethane in the composition is about 0.5 μM to about 10 μM, or about 1 μM to about 5 μM, or about 1.5 μM to about 2.5 μM; the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM; the concentration of diallyl trisulfide is about 1.0 μM to about 50 μM, or about 5.0 μM to about 40 μM, or about 15 μM to about 30 μM, or about 20 μM to about 30 μM.

[0174] In some embodiments, the at least two other compounds are dibromomethane and dipropyl disulfide, and the concentration of trihalomethanes other than dibromochloromethane in the composition is about 0.5 μM to about 10 μM, or about 1 μM to about 5 μM, or about 1.5 μM to about 2.5 μM; the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM; the concentration of dipropyl disulfide is about 10 μM to about 50 μM, or about 15 μM to about 40 μM, or about 20 μM to about 40 μM.

[0175] In some embodiments, the at least two other compounds are dibromomethane and naringenin, and the concentration of trihalomethanes other than dibromochloromethane in the composition is about 0.5 μM to about 10 μM, or about 1 μM to about 5 μM, or about 1.5 μM to about 2.5 μM; the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM; the concentration of naringenin is about 50 μM to about 150 μM, or about 75 μM to about 125 μM, or about 90 μM to about 110 μM.

[0176] In some embodiments, the at least two other compounds are dibromomethane and garlic oil, and the concentration of trihalomethanes other than dibromochloromethane in the composition is about 0.5 μM to about 10 μM, or about 1 μM to about 5 μM, or about 1.5 μM to about 2.5 μM; the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM; and the amount of garlic oil is about 1 ppm, or about 2 ppm, or about 3 ppm, or about 4 ppm, or about 5 ppm.

[0177] -Ratio of bromoform to at least one other compound(s)

[0178] In some embodiments, the concentration ratio of bromoform to at least one other compound is from about 1:1 to about 1:10000. For example, in some embodiments, the ratio of bromoform to at least one other compound is from about 1:50 to about 1:9000, or from about 1:100 to about 1:9000, or from about 1:500 to about 1:9000, or from about 1:50 to about 1:100, or from about 1:100 to about 1:1000, or from about 1:100 to about 1:500, or from about 1:500 to about 1:1500, or from about 1:1000 to about 1:3000, or from about 1:10 to about 1:100, or from about 1:1000 to about 1:3000, or from about 1:3000 to about 1:5000, or from about 1:7000 to about 1:9000. Or from about 1:8000 to about 1:8500.

[0179] In some embodiments, the at least one other compound is dibromochloromethane, and the concentration ratio of bromoform to dibromochloromethane is from about 1:1 to about 1:10, or from about 1:1 to about 1:5, or from about 1:1 to about 1:2, or about 1:1, or about 1:1.2, or about 1:1.3, or about 1:1.4, or about 1:1.5, or about 1:1.6, or about 1:1.7, or about 1:1.8, or about 1:1.9.

[0180] In some embodiments, the at least one other compound is selected from diallyl disulfide, diallyl trisulfide, dimethyl disulfide, and dipropyl disulfide, and the concentration ratio of bromoform to the compound is from about 1:100 to about 1:10000, e.g., from about 1:500 to about 1:10000, or from about 1:750 to about 1:1250, or from about 1:7500 to about 1:9000.

[0181] In some embodiments, the at least one other compound is diallyl disulfide, and the ratio of bromoform to the diallyl disulfide is about 1:100 to about 1:3000, such as about 1:100 to about 1:2500, or about 1:100 to about 1:2100.

[0182] In some embodiments, the at least one other compound is diallyl trisulfide, and the concentration ratio of bromoform to the diallyl trisulfide is about 1:500 to about 1:1500, such as about 1:750 to about 1:1250, or about 1:1000 to about 1:1100.

[0183] In some embodiments, the at least one other compound is dimethyl disulfide, and the concentration ratio of bromoform to dimethyl disulfide is about 1:7500 to 1:9000, such as about 1:8000 to 1:8500.

[0184] In some embodiments, the at least one other compound is dipropyl disulfide, and the concentration ratio of bromoform to dipropyl disulfide is about 1:7500 to about 1:9000, such as about 1:8000 to about 1:8500.

[0185] In some embodiments, the at least one additional compound is selected from prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sangsin, isopsoralen chalcone, naringenin, quercetin, and xanthohumol, and the concentration ratio of bromoform to the compound is about 1:10 to about 1:1000, e.g., about 1:25 to about 1:1000, or about 1:50 to about 1:100, about 1:50 to about 1:150.

[0186] In some embodiments, the at least one other compound is prenylnaringenin, and the concentration ratio of bromoform to the compound is about 1:50 to about 1:150, such as about 1:75 to about 1:125, or about 1:100 to about 1:110. In one embodiment, the prenylnaringenin is 6-prenylnaringenin or 8-prenylnaringenin, preferably 8-prenylnaringenin.

[0187] In some embodiments, the at least one other compound is psoralen, and the concentration ratio of bromoform to the compound is about 1:100 to 1:300, such as about 1:150 to about 1:250, or 1:190 to 1:210.

[0188] In some embodiments, the at least one other compound is psoralen, and the concentration ratio of bromoform to the compound is about 1:50 to 1:1000, such as about 1:100 to about 1:850, or 1:50 to 1:150, or about 1:75 to about 1:125, or about 1:100 to about 1:110.

[0189] In some embodiments, the at least one other compound is chalcone glycoside A or chalcone glycoside C, and the concentration ratio of bromoform to the compound is about 1:10 to about 1:50, such as about 1:15 to about 1:35, or about 1:20 to about 1:30. In one embodiment, the at least one other compound is chalcone glycoside A.

[0190] In some embodiments, the at least one other compound is sangrin, and the concentration ratio of bromoform to the compound is about 1:2 to about 1:100, such as about 1:5 to about 1:90, or about 1:10 to about 1:90, or about 1:10 to about 1:30, or about 1:15 to about 1:25.

[0191] In some embodiments, the at least one other compound is isopsoralea corylifolia chalcone, and the concentration ratio of bromoform to the compound is about 1:10 to about 1:100, e.g., about 1:15 to about 1:75, or about 1:20 to about 1:60, or about 1:40 to about 1:60, or about 1:45 to about 1:55.

[0192] In some embodiments, the at least one other compound is naringenin, and the concentration ratio of bromoform to the compound is about 1:500 to about 1:1500, such as about 1:650 to about 1:1000, or about 1:750 to about 1:900, or about 1:800 to about 1:850.

[0193] In some embodiments, the at least one other compound is quercetin, and the concentration ratio of bromoform to the compound is about 1:100 to about 1:1500, such as about 1:250 to about 1:1000, or about 1:350 to about 1:900, or about 1:750 to about 1:1000, or about 1:800 to about 1:850.

[0194] In some embodiments, the at least one other compound is xanthohumol, and the concentration ratio of bromoform to the compound is about 1:25 to about 1:150, such as about 1:40 to about 1:120, or about 1:80 to about 1:120, or about 1:100 to about 1:110.

[0195] In some embodiments, the at least one other compound is tannic acid, and the concentration ratio of bromoform to the compound is about 1:500 to about 1:1500, such as about 1:750 to about 1:1000, or about 1:800 to about 1:900, or about 1:800 to about 1:850.

[0196] In some embodiments, the at least one other compound is selected from cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil and lemon verbena oil, and the concentration ratio of bromoform to the amount of the compound is from about 1:100 to about 1:10000, for example, from about 1:200 to about 1:9000, or from about 1:500 to about 1:2500, or from about 1:3000 to about 1:5000, or from about 1:7500 to 1:10000, or from about 1:200 to about 1:300.

[0197] In some embodiments, the at least one other compound is cassia oil, and the concentration ratio of bromoform to the amount of the compound is from about 1:7500 to about 1:10000, or from about 1:8000 to about 1:9000, or from about 1:8000 to about 1:8500.

[0198] In some embodiments, the at least one other compound is garlic oil, and the concentration ratio of bromoform to the amount of the compound is from about 1:500 to about 1:4000, or from about 1:750 to about 1:3000, or from about 1:1000 to about 1:2500, or from about 1:1500 to about 1:2500, or from about 1:1750 to about 1:2250, or from about 1:2000 to about 1:2100.

[0199] In some embodiments, the at least one other compound is sandalwood oil, and the concentration ratio of bromoform to the amount of the compound is from about 1:100 to about 1:500, or from about 1:200 to about 1:400, or from about 1:200 to about 1:300, or from about 1:250 to about 1:270.

[0200] In some embodiments, the at least one other compound is thyme red oil, and the concentration ratio of bromoform to the amount of the compound is from about 1:2500 to about 1:7500, or from about 1:3000 to about 1:5000, or from about 1:3750 to about 1:4500, or from about 1:4000 to about 1:4500, or from about 1:4100 to about 1:4250, or from about 1:4100 to about 1:4200.

[0201] In some embodiments, the at least one other compound is lemongrass oil, and the concentration ratio of bromoform to the amount of the compound is from about 1:2500 to about 1:7500, or from about 1:3000 to about 1:5000, or from about 1:3750 to about 1:4500, or from about 1:4000 to about 1:4500, or from about 1:4100 to about 1:4250, or from about 1:4100 to about 1:4200.

[0202] In some embodiments, the at least one other compound is lemon verbena oil, and the concentration ratio of bromoform to the amount of the compound is from about 1:2500 to about 1:7500, or from about 1:3000 to about 1:5000, or from about 1:3750 to about 1:4500, or from about 1:4000 to about 1:4500, or from about 1:4100 to about 1:4250, or from about 1:4100 to about 1:4200.

[0203] In some embodiments, the at least one other compound is selected from carvacrol, citral, cinnamaldehyde, and hexylcinnamaldehyde (e.g., α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde), and the concentration ratio of bromoform to the compound is from about 1:200 to about 1:10000, for example, from about 1:200 to about 1:300, or from about 1:3000 to about 1:9000, or from about 1:3000 to about 1:5000, or from about 1:7500 to about 1:10000.

[0204] In some embodiments, the at least one other compound is carvacrol, and the concentration ratio of bromoform to the compound is about 1:2500 to about 1:7500, or about 1:3000 to about 1:5000, or about 1:3750 to about 1:4500, or about 1:4000 to about 1:4500, or about 1:4100 to about 1:4250, or about 1:4100 to about 1:4200.

[0205] In some embodiments, the at least one other compound is citral, and the concentration ratio of bromoform to the compound is about 1:2500 to about 1:7500, or about 1:3000 to about 1:5000, or about 1:3750 to about 1:4500, or about 1:4000 to about 1:4500, or about 1:4100 to about 1:4250, or about 1:4100 to about 1:4200.

[0206] In some embodiments, the at least one other compound is cinnamaldehyde, and the concentration ratio of bromoform to the compound is from about 1:7500 to about 1:10000, or from about 1:8000 to about 1:9000, or from about 1:8000 to about 1:8500.

[0207] In some embodiments, the at least one other compound is hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), and the concentration ratio of bromoform to the compound is from about 1:100 to about 1:500, or from about 1:200 to about 1:400, or from about 1:200 to about 1:300, or from about 1:250 to about 1:270. In one embodiment, the at least one other compound is α-hexyl cinnamaldehyde.

[0208] In some embodiments, the at least one other compound is selected from sclareol, saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C, or saikosaponin D), ursolic acid, and ginsenoside (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, or ginsenoside Rf), and the concentration ratio of bromoform to the compound is about 1:50 to about 1:1000, or about 1:50 to about 1:250, or about 1:750 to about 1:900.

[0209] In some embodiments, the at least one other compound is sclareol, and the concentration ratio of bromoform to the compound is from about 1:50 to about 1:500, or from about 1:100 to about 1:350, or from about 1:150 to about 1:250, or from about 1:175 to about 1:225.

[0210] In some embodiments, the at least one other compound is a saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C, or saikosaponin D), and the concentration ratio of bromoform to the compound is from about 1:25 to about 1:150, or from about 1:50 to about 1:100, or from about 1:70 to about 1:90, or from about 1:80 to about 1:85. In one embodiment, the at least one other compound is saikosaponin b2.

[0211] In some embodiments, the at least one other compound is ursolic acid, and the concentration ratio of bromoform to the compound is about 1:500 to about 1:1500, such as about 1:750 to about 1:1000, or about 1:800 to about 1:900, or about 1:800 to about 1:850.

[0212] In some embodiments, the at least one other compound is a ginsenoside (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, or ginsenoside Rf), and the concentration ratio of bromoform to the compound is from about 1:50 to about 1:350, or from about 1:50 to about 1:250, or from about 1:100 to about 1:200, or from about 1:150 to about 1:175. In one embodiment, the at least one other compound is ginsenoside Rb2.

[0213] In some embodiments, the at least one other compound is dibromoacetic acid, and the concentration ratio of bromoform to dibromoacetic acid is about 1:1000 to about 1:2000, such as about 1:1500 to about 1:1750, or about 1:1600 to about 1:1700.

[0214] In some embodiments, the at least one other compound is 3-nitrooxypropanol, and the concentration ratio of bromoform to 3-nitrooxypropanol is about 1:2 to about 1:20, e.g., about 1.5 to about 1:15, or about 1:5 to about 1:10, or about 1:6 to about 1:8.

[0215] Bromoform is generally unstable at relatively high concentrations and potentially toxic at such concentrations, so it is advantageous to obtain the effect of reducing methane production at the lowest possible concentration. According to the present invention, this can be achieved for bromoform as long as bromoform is used in combination with at least one other compound described herein. In fact, as demonstrated in the examples at very low concentrations, bromoform itself does not reduce or inhibit methane production. However, when combined with at least one other compound described herein at such low concentrations, a reduction or inhibition of methane production can be achieved.

[0216] In some embodiments, the concentration of bromoform in the composition is less than 0.30 μM, such as less than 0.25 μM, or less than 0.20 μM, or less than 0.15 μM. In some embodiments, the concentration of bromoform in the composition is at least about 0.05 μM, such as at least about 0.075 μM, or at least about 0.10 μM. In some embodiments, the concentration is about 0.05 μM to about 0.20 μM, such as about 0.075 μM to about 0.175 μM, or about 0.10 μM to about 0.15 μM, or about 0.11 μM to about 0.13 μM.

[0217] In some embodiments, the concentration of at least one other compound in the composition is higher than the concentration of bromoform. For example, in some embodiments, the concentration of at least one other compound is at least about 2.0 μM, such as at least about 10.0 μM, or at least about 50.0 μM, or at least about 100.0 μM, or at least about 250.0 μM, or at least about 500.0 μM, or at least about 750.0 μM. In some embodiments, the concentration of at least one other compound is less than about 5.0 mM, such as less than about 2.5 mM, or less than about 1.0 mM, or less than about 750.0 μM, or less than about 500.0 μM, or less than about 250.0 μM.

[0218] In some embodiments, the concentration of at least one other compound in the composition is about 0.15 μM to about 1.0 mM, for example, about 2.0 μM to about 1.0 mM, or about 2.0 μM to about 750.0 μM, or about 2.0 μM to about 500.0 μM, or about 2.0 μM to about 250.0 μM, or about 2.0 μM to about 150.0 μM, or about 2.0 μM to about 100.0 μM.

[0219] In some embodiments, the at least one other compound is dibromochloromethane, and the concentration of dibromochloromethane in the composition is about 0.15 μM to about 1.0 μM, such as about 0.15 μM to about 0.5 μM, or about 0.15 μM to about 0.25 μM.

[0220] In some embodiments, the at least one other compound is diallyl disulfide, and the concentration of diallyl disulfide in the composition is about 5.0 μM to about 500.0 μM, such as about 10.0 μM to about 350.0 μM, or about 15.0 μM to about 250.0 μM.

[0221] In some embodiments, the at least one other compound is diallyl trisulfide, and the concentration of diallyl trisulfide in the composition is about 50.0 μM to about 250.0 μM, such as about 75.0 μM to about 200.0 μM, or about 100.0 μM to about 150.0 μM.

[0222] In some embodiments, the at least one other compound is dimethyl disulfide, and the concentration of dimethyl disulfide in the composition is about 500.0 μM to about 1.5 mM, such as about 750 μM to about 1.25 mM, or about 900.0 μM to about 1.1 mM.

[0223] In some embodiments, the at least one other compound is dipropyl disulfide, and the concentration of dipropyl disulfide in the composition is about 500.0 μM to about 1.5 mM, such as about 750 μM to about 1.25 mM, or about 900.0 μM to about 1.1 mM.

[0224] In some embodiments, the at least one other compound is prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), and the concentration of prenylnaringenin in the composition is about 2.0 μM to about 50.0 μM, e.g., about 5.0 μM to about 25.0 μM, or about 10.0 μM to about 15.0 μM. In one embodiment, the prenylnaringenin is 8-prenylnaringenin.

[0225] In some embodiments, the at least one other compound is psoralen, and the concentration of psoralen in the composition is about 5.0 μM to about 75.0 μM, such as about 10.0 μM to about 50.0 μM, or about 15.0 μM to about 35.0 μM, or about 20.0 μM to about 30.0 μM.

[0226] In some embodiments, the at least one other compound is psoralen, and the concentration of psoralen in the composition is about 2.0 μM to about 200.0 μM, such as about 5.0 μM to about 150.0 μM, or about 10.0 μM to about 125.0 μM.

[0227] In some embodiments, the at least one other compound is chalcone glycoside A or chalcone glycoside C, and the concentration of chalcone glycoside A or chalcone glycoside C in the composition is about 0.5 μM to about 10.0 μM, such as about 1.0 μM to about 7.5 μM, or about 2.0 μM to about 5.0 μM. In one embodiment, the at least one other compound is chalcone glycoside A.

[0228] In some embodiments, the at least one other compound is sangsin, and the concentration of sangsin in the composition is about 0.25 μM to about 50.0 μM, such as about 0.50 μM to about 25.0 μM, or about 1.0 μM to about 15.0 μM.

[0229] In some embodiments, the at least one other compound is psoralea chalcone, and the concentration of psoralea chalcone in the composition is about 1.0 μM to about 15.0 μM, such as about 2.5 μM to about 12.5 μM, or about 4.0 μM to about 10.0 μM, or about 5.0 μM to about 7.5 μM.

[0230] In some embodiments, the at least one other compound is naringenin, and the concentration of naringenin in the composition is about 25.0 μM to about 250.0 μM, such as about 50.0 μM to about 150.0 μM, or about 75.0 μM to about 125.0 μM, or about 90.0 μM to about 110.0 μM.

[0231] In some embodiments, the at least one other compound is quercetin, and the concentration of quercetin in the composition is about 25.0 μM to about 250.0 μM, such as about 50.0 μM to about 150.0 μM, or about 75.0 μM to about 125.0 μM, or about 90.0 μM to about 110.0 μM.

[0232] In some embodiments, the at least one other compound is xanthohumol, and the concentration of xanthohumol in the composition is about 2.0 μM to about 50.0 μM, such as about 5.0 μM to about 25.0 μM, or about 10.0 μM to about 15.0 μM.

[0233] In some embodiments, the at least one other compound is tannic acid, and the concentration of tannic acid in the composition is about 25.0 μM to about 250.0 μM, such as about 50.0 μM to about 150.0 μM, or about 75.0 μM to about 125.0 μM, or about 90.0 μM to about 110.0 μM.

[0234] In some embodiments, the at least one other compound is carvacrol, and the concentration of carvacrol in the composition is about 250.0 μM to about 1.0 mM, such as about 250 μM to about 750.0 μM, or about 400.0 μM to about 600.0 μM, or about 450.0 μM to about 550.0 μM.

[0235] In some embodiments, the at least one other compound is citral, and the concentration of citral in the composition is about 250.0 μM to about 1.0 mM, such as about 250 μM to about 750.0 μM, or about 400.0 μM to about 600.0 μM, or about 450.0 μM to about 550.0 μM.

[0236] In some embodiments, the at least one other compound is cinnamaldehyde, and the concentration of cinnamaldehyde in the composition is about 500.0 μM to about 1.5 mM, such as about 750 μM to about 1.25 mM, or about 900.0 μM to about 1.1 mM.

[0237] In some embodiments, the at least one other compound is hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), and the concentration of hexyl cinnamaldehyde in the composition is about 10.0 μM to about 75.0 μM, such as about 15.0 μM to about 50.0 μM, or about 20.0 μM to about 40.0 μM, or about 25.0 μM to about 35.0 μM. In one embodiment, the at least one other compound is α-hexyl cinnamaldehyde.

[0238] In some embodiments, the at least one other compound is sclareol, and the concentration of sclareol in the composition is about 5.0 μM to about 75.0 μM, such as about 10.0 μM to about 50.0 μM, or about 15.0 μM to about 35.0 μM, or about 20.0 μM to about 30.0 μM.

[0239] In some embodiments, the at least one other compound is a saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C, or saikosaponin D), and the concentration of saikosaponin in the composition is about 2.0 μM to about 25.0 μM, such as about 5.0 μM to about 15.0 μM, or about 8.0 μM to about 12.0 μM. In one embodiment, the at least one other compound is saikosaponin b2.

[0240] In some embodiments, the at least one other compound is ursolic acid, and the concentration of ursolic acid in the composition is about 25.0 μM to about 250.0 μM, such as about 50.0 μM to about 150.0 μM, or about 75.0 μM to about 125.0 μM, or about 90.0 μM to about 110.0 μM.

[0241] In some embodiments, the at least one other compound is a ginsenoside (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, or ginsenoside Rf), and the concentration of ginsenosides in the composition is about 5.0 μM to about 50.0 μM, for example, about 10.0 μM to about 40.0 μM, or about 15.0 μM to about 30.0 μM, or about 15.0 μM to about 25.0 μM. In one embodiment, the at least one other compound is ginsenoside Rb2.

[0242] In some embodiments, the at least one other compound is dibromoacetic acid, and the concentration of dibromoacetic acid in the composition is about 50.0 μM to about 500.0 μM, such as about 100.0 μM to about 350.0 μM, or about 150.0 μM to about 250.0 μM, or about 175.0 μM to about 225.0 μM.

[0243] In some embodiments, the at least one other compound is 3-nitrooxypropanol, and the concentration of 3-nitrooxypropanol in the composition is about 0.25 μM to about 4.0 μM, for example, about 0.50 μM to about 2.5 μM, or about 0.50 μM to about 1.75 μM, or about 0.50 μM to about 1.5 μM, or about 0.50 μM to about 1.0 μM, or about 0.70 μM to about 0.85 μM.

[0244] In some embodiments, the at least one other compound is cassia oil, and the amount of cassia oil in the composition is from about 100 ppm to about 500 ppm, for example from about 150 ppm to about 350 ppm, or from about 200 ppm to about 300 ppm, or from about 225 ppm to about 275 ppm.

[0245] In some embodiments, the at least one other compound is garlic oil, and the amount of garlic oil in the composition is from about 10 ppm to about 250 ppm, such as from about 25 ppm to about 150 ppm, or from about 40 ppm to about 100 ppm, or from about 50 ppm to about 75 ppm.

[0246] In some embodiments, the at least one other compound is sandalwood oil, and the amount of sandalwood oil in the composition is from about 1 ppm to about 20 ppm, for example from about 3 ppm to about 15 ppm, or from about 4 ppm to about 10 ppm, or from about 6 ppm to about 8 ppm, or from about 7 ppm to about 8 ppm.

[0247] In some embodiments, the at least one other compound is thyme red oil, and the amount of thyme red oil in the composition is from about 10 ppm to about 250 ppm, such as from about 50 ppm to about 200 ppm, or from about 75 ppm to about 150 ppm, or from about 100 ppm to about 150 ppm.

[0248] In some embodiments, the at least one other compound is lemongrass oil, and the amount of lemongrass oil in the composition is from about 10 ppm to about 250 ppm, for example from about 50 ppm to about 200 ppm, or from about 75 ppm to about 150 ppm, or from about 100 ppm to about 150 ppm.

[0249] In some embodiments, the at least one other compound is lemon verbena oil, and the amount of lemon verbena oil in the composition is from about 10 ppm to about 250 ppm, such as from about 50 ppm to about 200 ppm, or from about 75 ppm to about 150 ppm, or from about 100 ppm to about 150 ppm.

[0250] -Ratio of iodoform to at least one other compound(s)

[0251] In some embodiments, the composition comprises iodoform and at least one other compound, for example, iodoform and one other compound.

[0252] In some embodiments, the concentration ratio of iodoform to at least one other compound is about 1:0.05 to about 1:10000. For example, in some embodiments, the concentration ratio of iodoform to at least one other compound is about 1:0.05 to about 1:100, or about 1:0.1 to about 1:75, or about 1:0.1 to about 1:50.

[0253] In some embodiments, the at least one other compound is dibromomethane, and the concentration ratio of iodoform to dibromomethane is from about 1:0.05 to about 1:1, or from about 1:0.1 to about 1:0.75, or from about 1:0.15 to about 1:0.5, or from about 1:0.2 to about 1:0.3.

[0254] In some embodiments, the at least one other compound is selected from diallyl disulfide, diallyl trisulfide, dimethyl disulfide, and dipropyl disulfide, and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20.

[0255] In some embodiments, the at least one other compound is diallyl disulfide, and the ratio of iodoform to the diallyl disulfide is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0256] In some embodiments, the at least one other compound is diallyl trisulfide, and the concentration ratio of iodoform to the diallyl trisulfide is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0257] In some embodiments, the at least one other compound is dipropyl disulfide, and the concentration ratio of iodoform to dipropyl disulfide is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:12.5 to about 1:20, or from about 1:12.5 to about 1:17.5.

[0258] In some embodiments, the at least one additional compound is selected from prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sangsin, isopsoralen chalcone, naringenin, quercetin, and xanthohumol, and the concentration ratio of iodoform to the compound is about 1:1 to about 1:500, or about 1:5 to about 1:250, or about 1:10 to about 1:100, or about 1:25 to about 1:75.

[0259] In some embodiments, the at least one other compound is naringenin, and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:500, or from about 1:5 to about 1:250, or from about 1:10 to about 1:100, or from about 1:25 to about 1:75, or from about 1:40 to about 1:60.

[0260] In some embodiments, the at least one other compound is selected from cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil, and lemon verbena oil, and the concentration ratio of iodoform to the amount of the compound is from about 1:0.5 to about 1:100, or from about 1:1 to about 1:75, or from about 1:2.5 to about 1:50, or from about 1:5 to about 1:30.

[0261] In some embodiments, the at least one other compound is garlic oil, and the concentration ratio of iodoform to the amount of the compound is from about 1:0.5 to about 1:100, from about 1:1 to about 1:50, or from about 1:2.5 to about 1:25 or from about 1:5 to about 1:10.

[0262] In some embodiments, the at least one other compound is thyme red oil, and the concentration ratio of iodoform to the amount of the compound is from about 1:1 to about 1:100, from about 1:2.5 to about 1:75, or from about 1:5 to about 1:50, or from about 1:10 to about 1:40, or from about 1:20 to about 1:30.

[0263] In some embodiments, the at least one other compound is lemongrass oil, and the concentration ratio of iodoform to the amount of the compound is from about 1:1 to about 1:100, from about 1:2.5 to about 1:75, or from about 1:5 to about 1:50, or from about 1:10 to about 1:40, or from about 1:20 to about 1:30.

[0264] In some embodiments, the at least one other compound is selected from carvacrol, citral, cinnamaldehyde, and hexylcinnamaldehyde (e.g., α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde), and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0265] In some embodiments, the at least one other compound is hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15. In one embodiment, the at least one other compound is α-hexyl cinnamaldehyde.

[0266] In some embodiments, the at least one other compound is pyromellitic acid imide, and the concentration ratio of iodoform to pyromellitic acid imide is from about 1:0.05 to about 1:25, or from about 1:0.1 to about 1:10, or from about 1:0.5 to about 1:5, or from about 1:1 to about 1:2.5.

[0267] In some embodiments, the composition comprises iodoform and at least two other compounds, such as iodoform and two other compounds. In some embodiments, the composition comprises iodoform and at least two other compounds, and the at least two other compounds are selected from dibromomethane, pyromellitic acid diimide, naringenin, dipropyl disulfide, garlic oil, diallyl disulfide, diallyl trisulfide, hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde), thyme red oil, lemongrass oil and any combination thereof. In some embodiments, the composition comprises iodoform, dibromomethane and one other compound, and the one other compound is selected from naringenin, dipropyl disulfide, garlic oil, diallyl disulfide or diallyl trisulfide.

[0268] In some embodiments, the concentration ratio of iodoform to the at least two other compounds is about 1:0.05:1 to about 1:1:500. For example, in some embodiments, the ratio of iodoform to the at least two other compounds is about 1:0.05:1 to about 1:0.75:250, or about 1:0.15:2 to about 1:0.5:100, or about 1:0.2:5 to about 1:0.3:50.

[0269] In some embodiments, one of the at least two other compounds is dibromomethane, and the concentration ratio of iodoform to dibromomethane is from about 1:0.05 to about 1:1, or from about 1:0.1 to about 1:0.75, or from about 1:0.15 to about 1:0.5, or from about 1:0.2 to about 1:0.3.

[0270] In some embodiments, one of the at least two other compounds is selected from diallyl disulfide, diallyl trisulfide, dimethyl disulfide, and dipropyl disulfide, and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20.

[0271] In some embodiments, one of the at least two other compounds is diallyl disulfide, and the ratio of iodoform to the diallyl disulfide is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0272] In some embodiments, one of the at least two other compounds is diallyl trisulfide, and the concentration ratio of iodoform to the diallyl trisulfide is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0273] In some embodiments, one of the at least two other compounds is dipropyl disulfide, and the concentration ratio of iodoform to dipropyl disulfide is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:12.5 to about 1:20, or from about 1:12.5 to about 1:17.5.

[0274] In some embodiments, one of the at least two additional compounds is selected from prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sangsin, isopsoralen chalcone, naringenin, quercetin, and xanthohumol, and the concentration ratio of iodoform to the compound is about 1:1 to about 1:500, or about 1:5 to about 1:250, or about 1:10 to about 1:100, or about 1:25 to about 1:75.

[0275] In some embodiments, one of the at least two other compounds is naringenin, and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:500, or from about 1:5 to about 1:250, or from about 1:10 to about 1:100, or from about 1:25 to about 1:75, or from about 1:40 to about 1:60.

[0276] In some embodiments, one of the at least two other compounds is selected from cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil and lemon verbena oil, and the concentration ratio of iodoform to the amount of the compound is from about 1:0.5 to about 1:100, or from about 1:1 to about 1:75, or from about 1:2.5 to about 1:50, or from about 1:5 to about 1:30.

[0277] In some embodiments, one of the at least two other compounds is garlic oil, and the concentration ratio of iodoform to the amount of the compound is from about 1:0.5 to about 1:100, from about 1:1 to about 1:50, or from about 1:2.5 to about 1:25 or from about 1:5 to about 1:10.

[0278] In some embodiments, one of the at least two other compounds is thyme red oil, and the concentration ratio of iodoform to the amount of the compound is from about 1:1 to about 1:100, from about 1:2.5 to about 1:75, or from about 1:5 to about 1:50, or from about 1:10 to about 1:40, from about 1:20 to about 1:30.

[0279] In some embodiments, one of the at least two other compounds is lemongrass oil, and the concentration ratio of iodoform to the amount of the compound is from about 1:1 to about 1:100, from about 1:2.5 to about 1:75, or from about 1:5 to about 1:50, or from about 1:10 to about 1:40, from about 1:20 to about 1:30.

[0280] In some embodiments, one of the at least two other compounds is selected from carvacrol, citral, cinnamaldehyde, and hexylcinnamaldehyde (e.g., α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde), and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15.

[0281] In some embodiments, one of the at least two other compounds is hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15. In one embodiment, the at least one other compound is α-hexyl cinnamaldehyde.

[0282] In some embodiments, one of the at least two other compounds is pyromellitic acid imide, and the concentration ratio of iodoform to pyromellitic acid imide is about 1:0.05 to about 1:25, or about 1:0.1 to about 1:10, or about 1:0.5 to about 1:5, or about 1:1 to about 1:2.5.

[0283] In some embodiments, the at least two other compounds are dibromomethane and diallyl disulfide, and the concentration of iodoform to dibromomethane to diallyl disulfide is from about 1:0.05:1 to about 1:1:50, or from about 1:0.1:5 to about 1:0.75:25, or from about 1:0.15:10 to about 1:0.5:20, or from about 1:0.2:10 to about 1:0.3:15.

[0284] In some embodiments, the at least two other compounds are dibromomethane and diallyl trisulfide, and the concentration of iodoform to dibromomethane to diallyl trisulfide is from about 1:0.05:1 to about 1:1:50, or from about 1:0.1:5 to about 1:0.75:25, or from about 1:0.15:10 to about 1:0.5:20, or from about 1:0.2:10 to about 1:0.3:15.

[0285] In some embodiments, the at least two other compounds are dibromomethane and dipropyl disulfide, and the concentration of iodoform to dibromomethane to dipropyl disulfide is about 1:0.05:1 to about 1:1:50, or about 1:0.1:5 to about 1:0.75:25, or about 1:0.15:10 to about 1:0.5:20, or about 1:0.2:12.5 to about 1:0.3:17.5.

[0286] In some embodiments, the at least two other compounds are dibromomethane and naringenin, and the concentration of iodoform to dibromomethane to naringenin is about 1:0.05:5 to about 1:1:250, or about 1:0.1:10 to about 1:0.75:100, or about 1:0.15:25 to about 1:0.5:75, or about 1:0.2:40 to about 1:0.3:60.

[0287] In some embodiments, the at least two other compounds are dibromomethane and garlic oil, and the concentration of iodoform to dibromomethane to garlic oil is about 1:0.05:0.5 to about 1:1:100, or about 1:0.1:1 to about 1:0.75:50, or about 1:0.15:2.5 to about 1:0.5:25, or about 1:0.2:5 to about 1:0.3:10.

[0288] Iodoform is generally more stable at relatively high concentrations and is less toxic than bromoform at these concentrations.

[0289] In some embodiments, the concentration of iodoform in the composition is at least about 0.5 μM, such as at least about 1 μM, or at least about 2 μM, or at least about 5 μM, or at least about 10 μM. In some embodiments, the concentration is about 0.5 μM to about 50 μM, or about 1 μM to about 25 μM, or about 1.5 μM to about 12.5 μM. In some embodiments, iodoform is about 1 μM to about 5 μM, such as 1.5 μM to about 2.5 μM.

[0290] In some embodiments, the concentration of the at least one other compound is at least about 0.5 μM, at least about 2.0 μM, such as at least about 10 μM, or at least about 50 μM, or at least about 100.0 μM, or at least about 250 μM, or at least about 500 μM, or at least about 750 μM. In some embodiments, the concentration of the at least one other compound is less than about 5.0 mM, such as less than about 2.5 mM, or less than about 1.0 mM, or less than about 750 μM, or less than about 500 μM, or less than about 250 μM, or less than about 100 μM, or less than about 50 μM, or less than about 10 μM, or less than about 5 μM.

[0291] In some embodiments, the concentration of at least one other compound in the composition is about 0.15 μM to about 1.0 mM, for example, about 0.5 μM to about 1.0 mM, or about 2.0 μM to about 750 μM, or about 2.0 μM to about 500 μM, or about 2.0 μM to about 250 μM, or about 2.0 μM to about 150 μM, or about 2.0 μM to about 100 μM.

[0292] In some embodiments, the at least one other compound is dibromomethane, and the concentration of dibromomethane in the composition is about 0.05 μM to about 50 μM, or about 0.1 μM to about 25 μM, or about 0.25 μM to about 12.5 μM.

[0293] In some embodiments, the at least one other compound is diallyl disulfide, and the concentration of diallyl disulfide in the composition is about 0.1 μM to about 100 μM, or about 0.5 μM to about 75 μM, or about 1.0 μM to about 50 μM, or about 5.0 μM to about 40 μM, or about 15 μM to about 30 μM.

[0294] In some embodiments, the at least one other compound is diallyl trisulfide, and the concentration of diallyl trisulfide in the composition is about 0.1 μM to about 100 μM, or about 0.5 μM to about 75 μM, or about 1.0 μM to about 50 μM, or about 5.0 μM to about 40 μM, or about 15 μM to about 30 μM.

[0295] In some embodiments, the at least one other compound is dipropyl disulfide, and the concentration of dipropyl disulfide in the composition is about 0.5 μM to about 100 μM, or about 1.0 μM to about 80 μM, or about 5.0 μM to about 60 μM, or about 10 μM to about 50 μM, or about 15 μM to about 40 μM, or about 20 μM to about 40 μM.

[0296] In some embodiments, the at least one other compound is naringenin, and the concentration of naringenin in the composition is about 25 μM to about 250 μM, such as about 50 μM to about 150 μM, or about 75 μM to about 125 μM, or about 90 μM to about 110 μM.

[0297] In some embodiments, the at least one other compound is hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), and the concentration of hexyl cinnamaldehyde in the composition is about 10 μM to about 75 μM, such as about 15 μM to about 50 μM, or about 20 μM to about 40 μM, or about 25 μM to about 35 μM. In one embodiment, the at least one other compound is α-hexyl cinnamaldehyde.

[0298] In some embodiments, the at least one other compound is garlic oil, and the amount of garlic oil in the composition is about 0.5 ppm, or about 1 ppm, or about 2 ppm, or about 3 ppm, or about 4 ppm, or about 5 ppm.

[0299] In some embodiments, the at least one other compound is thyme red oil, and the amount of thyme red oil in the composition is from about 0.5 ppm to about 50 ppm, or from about 1 ppm to about 40 ppm, or from about 5 ppm to about 35 ppm, or from about 10 ppm to about 30 ppm, or from about 15 ppm to about 25 ppm.

[0300] In some embodiments, the at least one other compound is lemongrass oil, and the amount of lemongrass oil in the composition is from about 0.5 ppm to about 50 ppm, or from about 1 ppm to about 40 ppm, or from about 5 ppm to about 35 ppm, or from about 10 ppm to about 30 ppm, or from about 15 ppm to about 25 ppm.

[0301] In some embodiments, the at least one other compound is pyromellitic acid imide, and the amount of pyromellitic acid imide in the composition is from about 0.1 ppm to about 10 ppm, or from about 0.25 ppm to about 7.5 ppm, or from about 0.5 to about 5 ppm, or from about 0.75 ppm to about 2.5 ppm.

[0302] In some embodiments, one of the at least two other compounds is dibromomethane, and the concentration of dibromomethane in the composition is about 0.05 μM to about 50 μM, or about 0.1 μM to about 25 μM, or about 0.25 μM to about 12.5 μM. In some embodiments, the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM.

[0303] In some embodiments, one of the at least two other compounds is diallyl disulfide, and the concentration of diallyl disulfide in the composition is from about 0.1 μM to about 100 μM, or from about 0.5 μM to about 75 μM, or from about 1.0 μM to about 50 μM, or from about 5.0 μM to about 40 μM, or from about 15 μM to about 30 μM.

[0304] In some embodiments, one of the at least two other compounds is diallyl trisulfide, and the concentration of diallyl trisulfide in the composition is from about 0.1 μM to about 100 μM, or from about 0.5 μM to about 75 μM, or from about 1.0 μM to about 50 μM, or from about 5.0 μM to about 40 μM, or from about 15 μM to about 30 μM.

[0305] In some embodiments, one of the at least two other compounds is dipropyl disulfide, and the concentration of dipropyl disulfide in the composition is about 0.5 μM to about 100 μM, or about 1.0 μM to about 80 μM, or about 5.0 μM to about 60 μM, or about 10 μM to about 50 μM, or about 15 μM to about 40 μM, or about 20 μM to about 40 μM.

[0306] In some embodiments, one of the at least two other compounds is naringenin, and the concentration of naringenin in the composition is about 25 μM to about 250 μM, such as about 50 μM to about 150 μM, or about 75 μM to about 125 μM, or about 90 μM to about 110 μM.

[0307] In some embodiments, one of the at least two other compounds is hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), and the concentration of hexyl cinnamaldehyde in the composition is about 10 μM to about 75 μM, such as about 15 μM to about 50 μM, or about 20 μM to about 40 μM, or about 25 μM to about 35 μM. In one embodiment, at least one other compound is α-hexyl cinnamaldehyde.

[0308] In some embodiments, one of the at least two other compounds is garlic oil, and the amount of garlic oil in the composition is about 0.5 ppm, or about 1 ppm, or about 2 ppm, or about 3 ppm, or about 4 ppm, or about 5 ppm.

[0309] In some embodiments, one of the at least two other compounds is thyme red oil, and the amount of thyme red oil in the composition is from about 0.5 ppm to about 50 ppm, or from about 1 ppm to about 40 ppm, or from about 5 ppm to about 35 ppm, or from about 10 ppm to about 30 ppm, or from about 15 ppm to about 25 ppm.

[0310] In some embodiments, one of the at least two other compounds is lemongrass oil, and the amount of lemongrass oil in the composition is from about 0.5 ppm to about 50 ppm, or from about 1 ppm to about 40 ppm, or from about 5 ppm to about 35 ppm, or from about 10 ppm to about 30 ppm, or from about 15 ppm to about 25 ppm.

[0311] In some embodiments, one of the at least two other compounds is pyromellitic acid imide, and the amount of pyromellitic acid imide in the composition is from about 0.1 ppm to about 10 ppm, or from about 0.25 ppm to about 7.5 ppm, or from about 0.5 to about 5 ppm, or from about 0.75 ppm to about 2.5 ppm.

[0312] In some embodiments, the at least two other compounds are dibromomethane and diallyl disulfide, and the concentration of iodoform in the composition is about 0.5 μM to about 10 μM, or about 1 μM to about 5 μM, or about 1.5 μM to about 2.5 μM; the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM; and the concentration of diallyl disulfide is about 1.0 μM to about 50 μM, or about 5.0 μM to about 40 μM, or about 15 μM to about 30 μM, or about 20 μM to about 30 μM.

[0313] In some embodiments, the at least two other compounds are dibromomethane and diallyl trisulfide, and the concentration of iodoform in the composition is about 0.5 μM to about 10 μM, or about 1 μM to about 5 μM, or about 1.5 μM to about 2.5 μM; the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM; and the concentration of diallyl trisulfide is about 1.0 μM to about 50 μM, or about 5.0 μM to about 40 μM, or about 15 μM to about 30 μM, or about 20 μM to about 30 μM.

[0314] In some embodiments, the at least two other compounds are dibromomethane and dipropyl disulfide, and the concentration of iodoform in the composition is about 0.5 μM to about 10 μM, or about 1 μM to about 5 μM, or about 1.5 μM to about 2.5 μM; the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM; and the concentration of dipropyl disulfide is about 10 μM to about 50 μM, or about 15 μM to about 40 μM, or about 20 μM to about 40 μM.

[0315] In some embodiments, the at least two other compounds are dibromomethane and naringenin, and the concentration of iodoform in the composition is about 0.5 μM to about 10 μM, or about 1 μM to about 5 μM, or about 1.5 μM to about 2.5 μM; the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM; and the concentration of naringenin is about 50 μM to about 150 μM, or about 75 μM to about 125 μM, or about 90 μM to about 110 μM.

[0316] In some embodiments, the at least two other compounds are dibromomethane and garlic oil, and the concentration of iodoform in the composition is about 0.5 μM to about 10 μM, or about 1 μM to about 5 μM, or about 1.5 μM to about 2.5 μM; the concentration of dibromomethane is about 0.1 μM to about 1 μM, for example, about 0.25 μM to about 0.75 μM; and the amount of garlic oil is about 1 ppm, or about 2 ppm, or about 3 ppm, or about 4 ppm, or about 5 ppm.

[0317] - Animal feed composition

[0318] The compositions disclosed herein, including all embodiments and any combination of embodiments, can be provided in the form of an animal feed composition or combined with an animal feed to form an animal feed composition.

[0319] For example, animal feed can be any composition suitable for providing nutrition to animals, particularly ruminants. For example, animal feed can be suitable for any ruminant, such as cattle, goats, sheep, yaks, deer or antelopes. Animal feed can be suitable for any pseudo-ruminant, such as camels and hippos. Animal feed can be suitable for any monogastric animal, such as kangaroos, rats, dogs, pigs, cats, horses, birds (such as pigeons, penguins) and rabbits. Animal feed can be suitable for monogastric herbivores, such as kangaroos, horses and rabbits.

[0320] Animal feed can be solid (e.g., powder, granules, pellets), semi-solid (e.g., gel, cream, paste) or liquid (e.g., solution, suspension, emulsion). The composition of the invention can be used, for example, for "top-dressing" (adding on top) animal feedyard rations or can be used for blending into a fully mixed ration. Animal feed can be added, for example, to the drinking water of the animal. In certain embodiments, the composition of the invention can be added to the drinking water of the animal immediately before ingestion, for example, up to 1 hour before ingestion, or up to 30 minutes before ingestion, or up to 15 minutes before ingestion, or up to 5 minutes before ingestion.

[0321] Three main types of animal feed include roughage, concentrate and mixed feed. Generally, roughage contains a high percentage of crude fiber and a low percentage of digestible nutrients. For example, roughage can be defined as containing equal to or greater than 20% by weight of crude fiber and equal to or less than 80% by weight of total digestible nutrients, such as equal to or greater than 20% by weight of crude fiber and equal to or less than 70% by weight of total digestible nutrients, equal to or greater than 20% by weight of crude fiber and equal to or less than 60% by weight of total digestible nutrients. Roughage can include, for example, dry roughage (e.g., hay, straw, artificial dehydrated feed, which contains 90% by weight of dry matter), silage (formed by green feed such as grass, alfalfa, sorghum and corn, and stored in silos with a dry matter content of 20 to 50%) and forage (e.g., green growing forage, which provides a feed with a high water content and only 20 to 30% of dry matter). Two basic types of roughage include grass and legumes. The fiber and dry matter of grass are generally higher than those of legumes. Legumes are generally higher in protein, energy, vitamins and minerals.

[0322] Concentrated feeds, in contrast to roughage, contain a low percentage of crude fiber and a high percentage of digestible nutrients. For example, concentrated feeds can be defined as containing less than 20% by weight of crude fiber and greater than 60% by weight of total digestible nutrients. Concentrated feeds can include, for example, energy-rich grains and molasses. Corn, oats, barley, and milo (sorghum grain) are energy-rich grains, containing about 70 to 80% by weight of total digestible nutrients. Concentrated feeds also include, for example, wheat bran, rice bran, wheat meal, rye meal, and rice flour.

[0323] Mixed feeds are generally a mixture of roughage and concentrate to provide a "complete" balanced ration, and may be high or low in energy, protein or fiber.

[0324] The composition of the present invention may be provided in the form of a concentrate feed for addition to an animal feed composition.

[0325] For example, a trihalomethane other than dibromochloromethane, such as bromoform or iodoform, and at least one other compound described herein can be combined with an animal feed in various amounts, depending on the total amount of the trihalomethane other than dibromochloromethane, such as bromoform or iodoform, and at least one other compound that is intended to be administered to or consumed by the animal.

[0326] The animal feed composition, for example, may comprise from about 0.0001% to about 10% by weight of the composition of the present invention, such as from about 0.01% to about 10% by weight, based on the total dry weight of the animal feed composition. For example, the animal feed composition may comprise from about 0.1% to about 9.5% by weight, or from about 0.4% to about 9.5% by weight, or from about 0.5% to about 9% by weight, or from about 0.6% to about 8.5% by weight, or from about 0.7% to about 8% by weight, or from about 0.8% to about 7.5% by weight, or from about 0.9% to about 7% by weight, or from about 1% to about 6% by weight, or from about 1.5% to about 5.5% by weight, or from about 2% to about 5% by weight, or from about 2.5% to about 4.5% by weight, or from about 3% to about 4% by weight of the composition of the present invention, based on the total dry weight of the animal feed composition. For example, the animal feed composition may comprise from about 0.0001 wt % to about 1.0 wt %, or from about 0.001 wt % to about 0.8 wt %, or from about 0.01 wt % to about 0.5 wt %, or from about 0.1 wt % to about 0.4 wt % of the composition of the present invention, based on the total dry weight of the animal feed composition.

[0327] For example, the animal feed composition can also include other animal feed supplements, including, for example, vitamins, minerals, antibiotics, growth stimulants and combinations thereof. For example, the animal feed composition can include other biologically active animal feed supplements, such as animal feed supplements suitable for reducing methane production / emission and / or increasing the availability of animal nutrients. Vitamins can be any one or more of vitamin A, vitamin D, vitamin E, vitamin K, thiamine, riboflavin, pyridoxine, cyanocobalamin, carotenoids (including beta-carotene, zeaxanthin, lutein and lycopene), niacin, folic acid, pantothenic acid, biotin, vitamin C, choline, inositol and salts and derivatives thereof. Minerals can be any one or more of calcium, phosphorus, magnesium, iron, zinc, manganese, copper, cobalt, boron, iodine, sodium, potassium, molybdenum, selenium, chromium, fluorine and chloride. 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.

[0328] Animal feed compositions may contain, in addition to active materials, other components such as flavoring agents, colorants, stabilizers (such as rosemary), enhancers (such as digestion enhancers), buffers, emulsifiers, dispersants, thickeners, solubilizers, trace nutrients (such as selenium), antioxidants (such as vitamins such as vitamin C, vitamin E), feed materials (such as carbohydrates, amino acids and urea), etc.

[0329] The animal feed composition may be in the form of, or provided as, an animal feed supplement, an animal feed additive or a complementary feed.

[0330] Animal feed supplements generally refer to any substance, preparation or product suitable for supplementing animal feed.

[0331] An animal feed additive is a substance, preparation or product that is intentionally added to feed (or water) to perform, in particular, one or more specific functions. For example, to meet the nutritional needs of the animal, or to improve the quality of the feed or to improve the performance and health of the animal. Animal feed additives are usually regulated products, i.e. they may only be placed on the market if they are authorised for use and only for the purposes specified in the authorisation.

[0332] A supplemental feed is a substance, preparation or product that is generally designed to be only part of an animal's diet and may not meet known nutritional requirements when fed alone. They are generally used in combination with additional food types described herein to meet daily requirements.

[0333] - Methods and uses

[0334] The compositions and animal feed compositions described herein (including all embodiments and combinations of embodiments) can be used to reduce methane production and / or emissions, increase the availability of nutrients for animals, and / or increase milk production and / or weight gain for animals and / or increase the efficiency of milk production and / or weight gain for animals. Without intending to be bound by theory, it is believed that these compositions can reduce methane production by microorganisms in the intestines of animals, thereby reducing the nutrients used by those microorganisms and increasing the availability of nutrients for animals. These compositions can improve feed digestion to increase the availability of nutrients for animals.

[0335] In certain embodiments, the animal is a ruminant. Ruminants include, for example, cattle, goats, sheep, yaks, deer or antelopes. In certain embodiments, the animal is a pseudo-ruminant. Pseudo-ruminants include, for example, camels and hippos. In certain embodiments, the animal is a monogastric animal. Monogastric animals include, for example, kangaroos, rats, dogs, pigs, cats, horses, birds (such as pigeons and penguins) and rabbits. In certain embodiments, the animal is a monogastric herbivore. Monogastric herbivores include, for example, kangaroos, horses and rabbits.

[0336] The compositions and animal feed compositions described herein can be administered orally to animals. For example, the compositions and animal feed compositions described herein can be administered daily to animals.

[0337] The amount of the animal feed composition given to an animal or eaten by an animal may be different according to the needs of the animal and the severity of animal methane production. The daily total amount of the animal feed composition given to an animal or eaten by an animal may be, for example, about 1g to about 1000g per day, or about 10g to 100g per day, or about 10g to about 1000g per day, or about 100g to about 1000g per day, or about 200g to about 1000g per day, or about 300g to about 700g per day, or about 400 to about 500g per day. The amount may be different according to the needs of the animal and the severity of animal methane production. For example, the amount of the animal feed composition given to an animal or eaten by an animal may be different according to the type of animal, the size of the animal, the age of the animal and / or the intestinal microbial group of the subject. Determining the appropriate amount of a specific subject belongs to the skill of this area. If desired, the daily total amount may be divided into several parts and given in one day (e.g., two or three parts per day). Generally, the suitable daily dose of an active agent will be the minimum effective dose to produce the desired effect. Increasing the amount of active agent (bromoform and at least one other compound described herein) administered to / consumed by an animal can, for example, reduce the amount of methane produced.

[0338] The compositions or animal feed compositions described herein can, for example, reduce methane production and / or emissions by at least about 1% (compared to methane production and / or emissions when the composition or animal feed composition is not consumed), for example, by at least 5% or at least 10%. For example, the compositions or animal feed compositions can reduce methane production and / or emissions by at least about 15%, or at least about 20%, 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%. The compositions or animal feed compositions described herein can, for example, reduce methane production and / or emissions by up to 100%. For example, the compositions or animal feed compositions can 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 evaluated, for example, according to the method described in the following examples.

[0339] The animal feed compositions described herein may, for example, increase milk production and / or weight gain by at least about 1% (compared to milk production and / or weight gain when the composition or animal feed composition is not consumed). For example, the composition or animal feed composition may increase milk production and / or weight gain by at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, 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%. The composition or animal feed composition described herein may, for example, increase milk production and / or weight gain by up to 100%. For example, the composition or animal feed composition may increase milk production and / or weight gain 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 the weight of the animal. Other measurement techniques include computed tomography (CT) scanning to measure empty body mass (total mass minus intestinal contents), carcass mass and composition analysis (lean muscle, fat and primary fat distribution; and changes in organs including the liver).

[0340] The compositions or animal feed compositions described herein can, for example, increase the efficiency of milk production and / or weight gain of an animal by at least about 1% (compared to the efficiency of milk production and / or weight gain of the animal when the composition or animal feed composition is not consumed). For example, the composition or animal feed composition can increase the efficiency of milk production and / or weight gain of an animal by at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, 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%. The compositions or animal feed compositions described herein can, for example, increase the efficiency of milk production and / or weight gain of an animal by up to 100%. For example, the composition or animal feed composition can increase the efficiency of milk production and / or weight gain of an animal 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, weight gain) occurs per unit of nutrition consumed. This can be measured, for example, by the change in the volume of milk produced per day or the animal's body weight divided by the total nutrients consumed by the animal.

[0341] As described herein, the components of the composition or animal feed composition can act synergistically to reduce methane production, inhibit methanogenesis and / or increase milk production and / or weight gain in animals. The synergistic combination of a trihalomethane other than dibromochloromethane, such as bromoform or iodoform, and at least one other compound described herein produces an effect greater than the sum of the effects of the individual components alone, thereby providing improved performance.

[0342] In some embodiments, the one or more methanogens are of the genus Methanobacterium, Methanosarcina, Methanobrevibacter, Methanosarcina, Methanoculleus, Methanosphaera, Methanocorpusculum, Methanofollis, Methanogenium, Methanomicrobium, Methanopyrus, Methanoregula, Methanasaeta, Methanthermobacter, or Methanococcus.In some embodiments, the one or more methanogens are selected from Methanobacterium formicicum, Methanobacterium bryantii, Methanobrevibacter ruminantium, Methanobrevibacter millerae, Methanobrevibacter olleyae, Methanomicrobium mobile, Methanoculleus olentangyi, Methanosarcina barkeri, Methanobrevibacter boviskoreani, Methanobacterium beijingense, Methanoculleus marisnigri, Methanoculleus bourgensis, Methanosarcina In some examples, the one or more methanogens are Methanococcus maripaludis.

[0343] - Manufacturing method

[0344] The composition described herein can be made by combining trihalomethanes other than dibromochloromethane, such as bromoform or iodoform, with at least one other compound described herein. The animal feed composition described herein can be made by combining animal feed with the composition described herein, i.e., trihalomethanes other than dibromochloromethane, such as bromoform or iodoform, and at least one other compound described herein. These components are combined in suitable amounts to obtain a composition of each component with a desired amount and / or concentration. Each component can be combined with one or more other components in any order and combination suitable for obtaining the desired product. For example, each component can be combined by mixing or blending, such as by surface addition. Such methods are well known in the art. The composition can be prepared into a dry solid form, such as a powder form, and is subjected to further processing steps according to the type of formulation of the expected final product. The method may further include a forming step, wherein the mixture is molded, pressed, spray-dried or otherwise formed into a shape (e.g., strips, balls, pellets, clusters, sheets), preferably with a size and / or texture suitable for animals of the type described herein to eat. The method may include containing the composition or animal feed composition in a specific delivery device such as a syringe. The method may include formulating the composition or animal feed composition into a bolus tablet that is intended to reside in the stomach of an animal (eg, the rumen of a ruminant).

[0345] The compositions and animal feed compositions described herein can be packaged in protective packaging materials, such as flexible intermediate bulk containers (FIBC), woven bags, paper bags, aluminum bags or polyethylene bags. The compositions and animal feed compositions can be packaged in boxes with waterproof and / or airtight materials or vacuum packed. The animal feed compositions can be packaged under nitrogen to prevent oxygen degradation.

[0346] The present disclosure may be described by one or more of the following paragraphs:

[0347] 1. A composition comprising bromoform and at least one other compound selected from the group consisting of dibromochloromethane, diallyl disulfide, diallyl trisulfide, dimethyl disulfide, dipropyl disulfide, isoprenylnaringenin (e.g., 6-isoprenylnaringenin or 8-isoprenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone A or chalcone C), sangsin, isopsoralen chalcone, naringenin, quercetin, xanthohumol, tannic acid, cinnamon oil, garlic oil , sandalwood oil, thyme red oil, lemongrass oil, lemon verbena oil, carvacrol, citral, cinnamaldehyde, hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), sclareol, saikosaponins (e.g., saikosaponin A, saikosaponin b2, saikosaponin C or saikosaponin D), ursolic acid, ginsenosides (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1 or ginsenoside Rf), dibromoacetic acid and 3-nitrooxypropanol and any combination thereof.

[0348] 2. The composition of paragraph 1, wherein the at least one other compound is dibromochloromethane.

[0349] 3. The composition according to paragraph 1, wherein the at least one other compound is selected from diallyl disulfide, diallyl trisulfide, dimethyl disulfide and dipropyl disulfide.

[0350] 4. The composition of paragraph 1, wherein the at least one additional compound is selected from prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone A or chalcone C), sangsin, isopsoralen chalcone, naringenin, quercetin, and xanthohumol.

[0351] 5. The composition according to paragraph 1, wherein the at least one other compound is tannic acid.

[0352] 6. The composition according to paragraph 1, wherein the at least one other compound is selected from cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil and lemon verbena oil.

[0353] 7. The composition according to paragraph 1, wherein the at least one other compound is selected from carvacrol, citral, cinnamaldehyde and hexylcinnamaldehyde (eg, α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde).

[0354] 8. The composition of paragraph 1, wherein the at least one other compound is selected from sclareol, saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C, or saikosaponin D), ursolic acid, and ginsenosides (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, or ginsenoside Rf).

[0355] 9. The composition of paragraph 1, wherein the at least one other compound is dibromoacetic acid.

[0356] 10. The composition of paragraph 1, wherein the at least one other compound is 3-nitrooxypropanol.

[0357] 11. The composition according to any of the preceding paragraphs, wherein the concentration ratio of bromoform to the at least one other compound is from about 1:1 to about 1:10000.

[0358] 12. A composition according to paragraph 1, wherein the compound:

[0359] (i) dibromochloromethane, and the concentration ratio of bromoform to dibromochloromethane is from 1:1 to 1:2; or

[0360] (ii) is selected from diallyl disulfide, diallyl trisulfide, dimethyl disulfide and dipropyl disulfide, and the concentration ratio of bromoform to the compound is from about 1:100 to about 1:10000, such as from about 1:500 to about 1:10000, or from about 1:750 to about 1:1250, or from about 1:7500 to about 1:9000; or

[0361] (iii) selected from 8-isoprenylnaringenin, psoralen, psoralen methyl ether, chalcone A, sangrin, isopsoralen chalcone, naringenin, quercetin and xanthohumol, and the concentration ratio of bromoform to the compound is from about 1:10 to about 1:1000, such as from about 1:25 to about 1:1000, or from about 1:50 to about 1:100, or from about 1:50 to about 1:150; or

[0362] (iv) tannic acid, and the concentration ratio of bromoform to tannic acid is from about 1:500 to about 1:1500, such as from about 1:750 to about 1:1000, or from about 1:800 to about 1:900; or

[0363] (v) selected from cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil and lemon verbena oil, and the concentration ratio of bromoform to the amount of the compound is from about 1:100 to about 1:10000, such as from about 1:200 to about 1:9000, or from about 1:500 to about 1:2500, or from about 1:3000 to about 1:5000, or from about 1:7500 to 1:10000, or from about 1:200 to about 1:300; or

[0364] (vi) is selected from carvacrol, citral, cinnamaldehyde and α-hexylcinnamaldehyde, and the concentration ratio of bromoform to the compound is from about 1:200 to about 1:10000, such as from about 1:200 to about 1:300, or from about 1:3000 to about 1:9000, or from about 1:3000 to about 1:5000, or from about 1:7500 to about 1:10000; or

[0365] (vii) selected from sclareol, saikosaponin b2, ursolic acid and ginsenoside Rb2, and the concentration ratio of bromoform to the compound is from about 1:50 to about 1:1000, or from about 1:50 to about 1:250, or from about 1:750 to about 1:900; or

[0366] (viii) is dibromoacetic acid, and the concentration ratio of bromoform to dibromoacetic acid is about 1:1000 to about 1:2000, such as about 1:1500 to about 1:1750; or

[0367] (ix) is 3-nitrooxypropanol, and the concentration ratio of bromoform to 3-nitrooxypropanol is about 1:2 to about 1:20, such as about 1:5 to about 1:15.

[0368] 13. A composition according to any preceding paragraph, wherein the concentration of bromoform in the composition is less than about 0.20 μM, such as less than about 0.15 μM, optionally wherein the concentration of bromoform in the composition is at least about 0.05 μM, such as at least about 0.10 μM.

[0369] 14. The composition of any preceding paragraph, wherein the at least one other compound is present at a concentration of about 1 μM to about 1.5 mM, for example, dibromochloromethane, diallyl disulfide, diallyl trisulfide, dimethyl disulfide and dipropyl disulfide, prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sansin, isopsoralen chalcone, naringenin, succinate ... For quercetin, quercetin, xanthohumol, tannic acid, carvacrol, citral, cinnamaldehyde, hexylcinnamaldehyde (e.g., α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde), sclareol, saikosaponins (e.g., saikosaponin A, saikosaponin b2, saikosaponin C, or saikosaponin D), ursolic acid, ginsenosides (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, or ginsenoside Rf), dibromoacetic acid, and 3-nitrooxypropanol, the range is about 10 μM to about 1 mM.

[0370] 15. The composition of any one of claims 1-13, wherein the amount of at least one other compound in the composition is from about 10 ppm to about 500 ppm, for example, from about 10 ppm to about 250 ppm for cinnamon oil, garlic oil, sandalwood oil, red thyme oil, lemongrass oil, and lemon verbena oil.

[0371] 16. An animal feed comprising the composition of any one of paragraphs 1-15, for example, wherein the animal feed is an animal feed supplement, or an animal feed additive, or a supplemental feed.

[0372] 17. Use of the composition according to paragraphs 1 to 15 or the animal feed according to paragraph 16 for:

[0373] a. Reducing methane emissions from animals; and / or

[0374] b. inhibiting methane production by one or more methanogens in an animal; and / or

[0375] c. Increase milk production and / or weight gain in animals.

[0376] 18. The use according to paragraph 17, wherein the composition is consumed by an animal.

[0377] 19. The use according to paragraph 17 or 18, wherein the animal is a ruminant, such as a cow, goat or sheep.

[0378] 20. A method comprising:

[0379] a. Reduce methane emissions from animals, and / or

[0380] b. inhibiting the production of methane by one or more methanogens in an animal, and / or

[0381] c. Increase milk production and / or weight gain in animals,

[0382] The method comprises administering to an animal a composition according to any one of paragraphs 1-15 or an animal feed according to paragraph 16.

[0383] 21. The method of paragraph 20, wherein the animal is a ruminant, such as a cow, goat or sheep.

[0384] 22. A composition comprising iodoform and at least one other compound selected from the group consisting of dibromochloromethane, diallyl disulfide, diallyl trisulfide, dimethyl disulfide, dipropyl disulfide, isoprenylnaringenin (e.g., 6-isoprenylnaringenin or 8-isoprenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone A or chalcone C), sangsin, isopsoralen chalcone, naringenin, quercetin, xanthohumol, tannic acid, cinnamon oil, garlic Oil, sandalwood oil, thyme red oil, lemongrass oil, lemon verbena oil, carvacrol, citral, cinnamaldehyde, hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), sclareol, saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C or saikosaponin D), ursolic acid, ginsenosides (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1 or ginsenoside Rf), dibromoacetic acid and 3-nitrooxypropanol, and any combination thereof.

[0385] 23. The composition of paragraph 22, wherein the at least one other compound is dibromomethane.

[0386] 24. The composition of paragraph 22, wherein the at least one other compound is selected from diallyl disulfide, diallyl trisulfide and dipropyl disulfide.

[0387] 25. The composition of paragraph 22, wherein the at least one other compound is naringenin.

[0388] 26. The composition of paragraph 22, wherein the at least one other compound is selected from garlic oil, thyme red oil, and lemongrass oil.

[0389] 27. The composition of paragraph 22, wherein the at least one other compound is hexylcinnamaldehyde, such as α-hexylcinnamaldehyde.

[0390] 28. The composition of paragraph 22, wherein the at least one other compound is pyromellitic acid diimide.

[0391] 29. The composition of any of paragraphs 22-28, wherein the concentration ratio of iodoform to the at least one other compound is from about 1:0.05 to about 1:10000.

[0392] 30. The composition of paragraph 22, wherein the composition comprises iodoform and at least two other compounds selected from the group consisting of dibromomethane, pyromellitic acid diimide, naringenin, dipropyl disulfide, garlic oil, diallyl disulfide, diallyl trisulfide, hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde), thyme red oil, lemongrass oil, and any combination thereof.

[0393] 31. A composition according to paragraph 30, wherein the at least two other compounds are:

[0394] (i) dibromomethane, and

[0395] (ii) naringenin, dipropyl disulfide, garlic oil, diallyl disulfide or diallyl trisulfide.

[0396] 32. The composition according to paragraphs 22-31, wherein the compound:

[0397] (i) is dibromomethane, and the concentration ratio of iodoform to dibromomethane is from about 1:0.05 to about 1:1, or from about 1:0.1 to about 1:0.75, or from about 1:0.15 to about 1:0.5, or from about 1:0.2 to about 1:0.3; and / or

[0398] (ii) is selected from diallyl disulfide, diallyl trisulfide and dipropyl disulfide, and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20; or

[0399] (iii) is naringenin, and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:500, or from about 1:5 to about 1:250, or from about 1:10 to about 1:100, or from about 1:25 to about 1:75, or from about 1:40 to about 1:60; or

[0400] (iv) selected from garlic oil, red thyme oil and lemongrass oil, and the concentration ratio of iodoform to the amount of the compound is from about 1:0.5 to about 1:100, or from about 1:1 to about 1:75, or from about 1:2.5 to about 1:50, or from about 1:5 to about 1:30; or

[0401] (v) is α-hexylcinnamaldehyde, and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15; or

[0402] (vi) is pyromellitic acid imide, and the concentration ratio of iodoform to pyromellitic acid imide is from about 1:0.05 to about 1:25, or from about 1:0.1 to about 1:10, or from about 1:0.5 to about 1:5, or from about 1:1 to about 1:2.5.

[0403] 33. The composition according to paragraphs 22-31, wherein the concentration of iodoform in the composition is about 1 μM to about 25 μM, such as about 1.5 μM to about 12.5 μM.

[0404] 34. The composition of paragraphs 22-31, wherein the concentration of the at least one other compound is about 0.05 μM to about 50 μM, such as about 0.25 μM to about 12.5 μM, for dibromomethane; about 0.1 μM to about 100.0 μM, such as about 15.0 μM to about 30.0 μM, for diallyl disulfide and diallyl trisulfide; about 0.5 μM to about 100.0 μM, such as about 20.0 μM to about 40.0 μM, for dipropyl disulfide; about 25.0 μM to about 250.0 μM, such as about 90.0 μM to about 110.0 μM, for naringenin; and about 10.0 μM to about 75.0 μM, such as about 25.0 μM to about 35.0 μM, for α-hexylcinnamaldehyde.

[0405] 35. The composition of paragraphs 22-31, wherein the amount of at least one other compound in the composition is about 0.5 ppm, such as about 5 ppm, for garlic oil; about 0.5 ppm to about 50 ppm, such as about 15 ppm to about 25 ppm, for thyme red oil and lemongrass oil; and about 0.1 ppm to about 10 ppm, such as about 0.75 ppm to about 2.5 ppm, for pyromellitic acid diimide.

[0406] 36. An animal feed comprising the composition of any one of paragraphs 22-35, for example, wherein the animal feed is an animal feed supplement, or an animal feed additive, or a supplemental feed.

[0407] 37. Use of the composition according to paragraphs 22-35 or the animal feed according to paragraph 36 for:

[0408] a. Reducing methane emissions from animals; and / or

[0409] b. inhibiting methane production by one or more methanogens in an animal; and / or

[0410] c. Increase milk production and / or weight gain in animals.

[0411] 38. The use according to paragraph 37, wherein the composition is consumed by an animal.

[0412] 39. The use according to paragraph 37 or 38, wherein the animal is a ruminant, such as a cattle, goat or sheep.

[0413] 40. A method comprising:

[0414] a. Reduce methane emissions from animals, and / or

[0415] b. inhibiting the production of methane by one or more methanogens in an animal, and / or

[0416] c. Increase milk production and / or weight gain in animals,

[0417] The method comprises administering to an animal a composition according to any one of paragraphs 1-35 or an animal feed according to paragraph 36.

[0418] 41. The method of paragraph 40, wherein the animal is a ruminant, such as a cow, goat or sheep.

[0419] The invention will now be described by reference only to the following non-limiting examples. Example

[0420] Example 1 - Inhibition of methane production by the methanogenic archaeon Methanococcus maripaludis using combinations of bromoform and various combinations of at least one other compound (each referred to as Compound X).

[0421] For this experiment, 100 mM bromoform was prepared by adding 8.75 μl bromoform to 991 μl DMSO. This was diluted 10-fold to give a 10 mM solution and further diluted to give a 0.12 mM stock solution. Compound X stock solutions were prepared in DMSO (except for tannic acid, which was diluted in deionized water) to obtain the desired stock solutions required for the test. The experiment was set up as follows by adding 5 ml of M141 medium (https: / / www.dsmz.de / microorganisms / medium / pdf / DSMZ_Medium141.pdf) to a screw cap Hungate tube and then adding 5 μl Compound X and / or 5 μl bromoform or 5 / 10 μl DMSO:

[0422] Tube 1: DMSO (10 μl)

[0423] Tube 2: Bromoform (5 μl, 0.120 mM) (final 120 nM) + 5 μl DMSO

[0424] Tube 3: Compound X (5 μl stock solution) + 5 μl DMSO

[0425] Tube 4: Bromoform (5 μl 0.120 mM) (final 120 nM) + Compound X (5 μl stock solution)

[0426] The concentration of bromoform was fixed at 0.12 μM. This concentration was too low to inhibit methanogens alone.

[0427] After adding the test substances, 500 μl of an overnight M. maritimus culture was added to each reaction tube. Each tube was aerated with 80% H2 / 20% CO2 to 240 kPa and incubated at 37°C for 24 hours.

[0428] After 24 hours of incubation, the pressure inside the tube was measured using a manometer. Assuming that 5 moles of H2 / CO2 are consumed to produce 1 mole of CH4, the observed pressure drop was used to calculate the amount of methane for the control and test reactions and, from this, the percent inhibition was calculated.

[0429] Table 1 below summarizes the concentration range of Compound X when tested with bromoform and the fold increase in inhibition of Compound X when tested with bromoform relative to the level of inhibition of Compound X when tested alone.

[0430]

[0431]

[0432] Example 2 - Inhibition of methane production by combination of iodoform and at least one other compound

[0433] The potential for methane production was evaluated for various samples of single species, extracts, two-species combinations and three-species combinations. The evaluation was carried out using an in vitro fermentation assay called a gas test. In each gas test, a 250mg total dietary ration (TMR) containing 30% grass, 30% corn and 40% complete feed was introduced into a 100ml serum bottle as a substrate for the fermentation process. The rumen fluid obtained from the fistulated cow of the University of Reading was mixed with a degassed reaction buffer (1.6L) and supplemented with Na2S (82ml). Subsequently, 30ml rumen fluid / buffer mixture was added to each serum bottle.

[0434] The test substances were dissolved in 100% ethanol and 100 μl of each test substance was added to the serum vials containing the rumen fluid / buffer mixture. For each experiment, all test substances and controls were checked in triplicate. Once the test substances and controls were added, the vials were sealed with butyl rubber stoppers and aluminum glands and then incubated at 39°C for 24 hours. After incubation, gas samples were collected from both the control and test vials and methane levels were determined using gas chromatography.

[0435] For each test compound, the “percent inhibition of methane produced” was calculated using the following formula:

[0436] Inhibition % = 100-(methane 测试 / Methane 对照 ×100)

[0437] The test substances were evaluated in three separate gas tests and the average inhibition percentage was calculated for each substance.

[0438] Two species combination

[0439] The synergistic effects between substances were determined using the Bliss independence model using the following formula:

[0440] I AB =I A +I B -I A I B

[0441] Among them I AB is the predicted methane inhibition percentage for the two substances together, and I A and I B is the observed methane inhibition percentage for a single substance. The predicted methane inhibition percentage for both substances (I AB ) and the observed methane inhibition percentages for both substances (I ob ) Calculate the combination index (Ci):

[0442] Ci=I AB / I ob

[0443] If Ci<1, the two substances have a synergistic effect.

[0444] Three species combination

[0445] Use the following calculation to determine the synergy between substances:

[0446] I ABC =I A +I B +I C-I A I B --I A I C -I B I C -I A I B I C

[0447] Among them I ABC is the predicted methane suppression percentage of the three substances together, and I A ,I B and I C is the observed percentage of methane suppression for the individual substances.

[0448] The predicted methane inhibition percentages (I ABC ) and the observed methane inhibition percentage for the combination of the three substances (I ob ) Calculate the combination index (Ci):

[0449] Ci=I ABC / I ob

[0450] If Ci<1, the two substances have a synergistic effect.

[0451] Table 1 shows the observed methane inhibition for individual species.

[0452] Table 2 shows the methane inhibition by the two-species combinations and the Ci value for each combination.

[0453] Table 3 shows the methane suppression of the three-species combinations and the Ci value for each combination.

[0454] Corresponding graphs illustrating methane suppression by single species, two-species combinations, and three-species combinations can be found in the Figures section.

[0455] Table 2: Methane inhibition by single compounds

[0456] Compound Mean observed methane suppression (%) Iodoform (2 μM) 40.8 Iodoform (10 μM) 98.3 Dibromomethane (0.5 μM) 14.5 Dibromomethane (10 μM) 99 Pyromellitic acid diimide (1μg / ml) 20 Naringenin (100μM) 10 Dipropyl disulfide (30 μM) 3 Garlic oil (5μg / ml) 1 Diallyl trisulfide (25 μM) 11 Diallyl disulfide (25 μM) 5 α-Hexylcinnamaldehyde (25μM) 8 Thyme red oil (20μg / ml) 5 Lemongrass oil (20 μg / ml) 5 Bromoform 0.1 μM 1.3 Bromoform 1 μM 92.4

[0457] Table 3: Methane suppression by two-species combination

[0458]

[0459] Table 4: Methane suppression by three-species combination

[0460]

[0461] The results obtained from the combination of iodoform with both pure compounds and plant extracts / oils indicated a significant synergistic effect in terms of methane inhibition, providing valuable insights for the development of effective strategies to mitigate methane emissions.

[0462] Synergistic effects were observed when iodoform was combined with specific pure compounds, such as dibromomethane, pyromellitic acid diimide, DPDS, DATS, α-hexylcinnamaldehyde, and naringenin, as well as plant extracts / oils, such as garlic oil, thyme oil, and lemongrass oil. This suggests that utilizing these combinations could result in stronger methane inhibition than using these species alone. Furthermore, the synergistic interactions suggest that it may be possible to reduce the amount of each compound required, thereby improving the cost-effectiveness of methane mitigation strategies.

[0463] The identification of synergistic interactions provides opportunities for the development of sustainable approaches for reducing livestock methane emissions. These combinations may help achieve stronger methane inhibition while potentially minimizing the amount of each compound required. This has important implications for optimizing the effectiveness and resource utilization of methane reduction strategies.

[0464] The implementation of these synergistic combinations is expected to promote environmentally friendly practices in the agricultural sector, promote sustainable livestock production, and mitigate the environmental impacts of methane emissions.

Claims

1. A composition comprising a trihalomethane other than dibromochloromethane and at least one other compound selected from the group consisting of dibromochloromethane, dibromomethane, diallyl disulfide, diallyl trisulfide, dimethyl disulfide, dipropyl disulfide, isoprenylnaringenin (e.g., 6-isoprenylnaringenin or 8-isoprenylnaringenin), bavachin, bavachinin, corylifol (e.g., corylifol A or corylifol C), sangsin, isobavachinol chalcone (e.g., isobavachinin), one), naringenin, quercetin, xanthohumol, tannic acid, cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil, lemon verbena oil, carvacrol, citral, cinnamaldehyde, hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde or 2-hexyl cinnamaldehyde), sclareol, saikosaponin (e.g., saikosaponin A, saikosaponin b2, saikosaponin C or saikosaponin D), ursolic acid, ginsenosides (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1 or ginsenoside Rf), dibromoacetic acid and 3-nitrooxypropanol, pyromellitic acid diimide and any combination thereof.

2. The composition of claim 1, wherein the at least one other compound is dibromochloromethane.

3. The composition of claim 1, wherein the at least one other compound is dibromomethane.

4. The composition of claim 1, wherein the at least one other compound is selected from diallyl disulfide, diallyl trisulfide, dimethyl disulfide and dipropyl disulfide.

5. The composition of claim 1, wherein the at least one additional compound is selected from prenylnaringenin (e.g., 6-prenylnaringenin or 8-prenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sangsin, isopsoralen chalcone, naringenin, quercetin, and xanthohumol.

6. The composition of claim 1, wherein the at least one other compound is tannic acid.

7. The composition according to claim 1, wherein the at least one other compound is selected from cinnamon oil, garlic oil, sandalwood oil, red thyme oil, lemongrass oil and lemon verbena oil.

8. The composition of claim 1, wherein the at least one additional compound is selected from carvacrol, citral, cinnamaldehyde, and hexylcinnamaldehyde (eg, α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde).

9. The composition of claim 1, wherein the at least one other compound is selected from sclareol, saikosaponins (e.g., saikosaponin A, saikosaponin b2, saikosaponin C, or saikosaponin D), ursolic acid, and ginsenosides (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, or ginsenoside Rf).

10. The composition of claim 1, wherein the at least one other compound is dibromoacetic acid.

11. The composition of claim 1, wherein the at least one other compound is 3-nitrooxypropanol.

12. The composition of claim 1, wherein the at least one other compound is pyromellitic acid diimide.

13. The composition of any one of the preceding claims, wherein the concentration ratio of trihalomethane to the at least one other compound is from about 1:0.05 to about 1:10000.

14. A composition according to any preceding claim, wherein the trihalomethane is bromoform.

15. The composition of any one of claims 1-13, wherein the trihalomethane is iodoform.

16. The composition of claim 15, wherein the composition comprises iodoform and at least two other compounds selected from the group consisting of dibromomethane, pyromellitic acid diimide, naringenin, dipropyl disulfide, garlic oil, diallyl disulfide, diallyl trisulfide, hexyl cinnamaldehyde (e.g., α-hexyl cinnamaldehyde), thyme red oil, lemongrass oil, and any combination thereof.

17. The composition of claim 16, wherein the at least two other compounds are: (i) dibromomethane and (ii) naringenin, dipropyl disulfide, garlic oil, diallyl disulfide or diallyl trisulfide.

18. The composition of claim 14, wherein the compound: (i) dibromochloromethane, and the concentration ratio of bromoform to dibromochloromethane is from 1:1 to 1:2; or (ii) is selected from diallyl disulfide, diallyl trisulfide, dimethyl disulfide and dipropyl disulfide, and the concentration ratio of bromoform to the compound is from about 1:100 to about 1:10000, such as from about 1:500 to about 1:10000, or from about 1:750 to about 1:1250, or from about 1:7500 to about 1:9000; or (iii) selected from 8-isoprenylnaringenin, psoralen, psoralen methyl ether, chalcone A, sangrin, isopsoralen chalcone, naringenin, quercetin and xanthohumol, and the concentration ratio of bromoform to the compound is from about 1:10 to about 1:1000, such as from about 1:25 to about 1:1000, or from about 1:50 to about 1:100, or from about 1:50 to about 1:150; or (iv) is tannic acid, and the concentration ratio of bromoform to tannic acid is about 1:500 to about 1:1500, such as about 1:750 to about 1:1000, or about 1:800 to about 1:900; or (v) selected from cinnamon oil, garlic oil, sandalwood oil, thyme red oil, lemongrass oil and lemon verbena oil, and the concentration ratio of bromoform to the amount of the compound is from about 1:100 to about 1:10000, such as from about 1:200 to about 1:9000, or from about 1:500 to about 1:2500, or from about 1:3000 to about 1:5000, or from about 1:7500 to 1:10000, or from about 1:200 to about 1:300; or (vi) is selected from carvacrol, citral, cinnamaldehyde and α-hexylcinnamaldehyde, and the concentration ratio of bromoform to the compound is from about 1:200 to about 1:10000, such as from about 1:200 to about 1:300, or from about 1:3000 to about 1:9000, or from about 1:3000 to about 1:5000, or from about 1:7500 to about 1:10000; or (vii) selected from sclareol, saikosaponin b2, ursolic acid and ginsenoside Rb2, and the concentration ratio of bromoform to the compound is from about 1:50 to about 1:1000, or from about 1:50 to about 1:250, or from about 1:750 to about 1:900; or (viii) is dibromoacetic acid, and the concentration ratio of bromoform to dibromoacetic acid is about 1:1000 to about 1:2000, such as about 1:1500 to about 1:1750; or (ix) is 3-nitrooxypropanol, and the concentration ratio of bromoform to 3-nitrooxypropanol is about 1:2 to about 1:20, such as about 1:5 to about 1:

15.

19. A composition according to claims 15 to 17, wherein the compound: (i) is dibromomethane, and the concentration ratio of iodoform to dibromomethane is from about 1:0.05 to about 1:1, or from about 1:0.1 to about 1:0.75, or from about 1:0.15 to about 1:0.5, or from about 1:0.2 to about 1:0.3; and / or (ii) is selected from diallyl disulfide, diallyl trisulfide and dipropyl disulfide, and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20; or (iii) is naringenin, and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:500, or from about 1:5 to about 1:250, or from about 1:10 to about 1:100, or from about 1:25 to about 1:75, or from about 1:40 to about 1:60; or (iv) selected from garlic oil, red thyme oil and lemongrass oil, and the concentration ratio of iodoform to the amount of the compound is from about 1:0.5 to about 1:100, or from about 1:1 to about 1:75, or from about 1:2.5 to about 1:50, or from about 1:5 to about 1:30; or (v) is α-hexylcinnamaldehyde, and the concentration ratio of iodoform to the compound is from about 1:1 to about 1:100, or from about 1:5 to about 1:50, or from about 1:10 to about 1:25, or from about 1:10 to about 1:20, or from about 1:10 to about 1:15; or (vi) is pyromellitic acid imide, and the concentration ratio of iodoform to pyromellitic acid imide is from about 1:0.05 to about 1:25, or from about 1:0.1 to about 1:10, or from about 1:0.5 to about 1:5, or from about 1:1 to about 1:2.

5.

20. The composition of claim 14, wherein the concentration of bromoform in the composition is less than about 0.20 μM, such as less than about 0.15 μM, optionally wherein the concentration of bromoform in the composition is at least about 0.05 μM, such as at least about 0.10 μM.

21. The composition according to claim 14, wherein The concentration of the at least one other compound is about 1 μM to about 1.5 mM, for example, dibromochloromethane, diallyl disulfide, diallyl trisulfide, dimethyl disulfide and dipropyl disulfide, isoprenylnaringenin (e.g., 6-isoprenylnaringenin or 8-isoprenylnaringenin), psoralen, psoralen methyl ether, chalcone glycosides (e.g., chalcone glycosides A or chalcone glycosides C), sangsin, isopsoralen chalcone, naringenin, quercetin, xanthohumol For , tannic acid, carvacrol, citral, cinnamaldehyde, hexylcinnamaldehyde (e.g., α-hexylcinnamaldehyde or 2-hexylcinnamaldehyde), sclareol, saikosaponins (e.g., saikosaponin A, saikosaponin b2, saikosaponin C or saikosaponin D), ursolic acid, ginsenosides (e.g., ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1 or ginsenoside Rf), dibromoacetic acid, and 3-nitrooxypropanol, it is about 10 μM to about 1 mM.

22. The composition of claim 14, wherein the amount of at least one other compound in the composition is from about 10 ppm to about 500 ppm, for example, from about 10 ppm to about 250 ppm for cinnamon oil, garlic oil, sandalwood oil, red thyme oil, lemongrass oil, and lemon verbena oil.

23. The composition of claims 15-17, wherein the concentration of iodoform in the composition is about 1 μM to about 25 μM, such as about 1.5 μM to about 12.5 μM.

24. The composition of claims 15-17, wherein the concentration of the at least one other compound is from about 0.05 μM to about 50 μM, such as from about 0.25 μM to about 12.5 μM, for dibromomethane; from about 0.1 μM to about 100.0 μM, such as from about 15.0 μM to about 30.0 μM, for diallyl disulfide and diallyl trisulfide; from about 0.5 μM to about 100.0 μM, such as from about 20.0 μM to about 40.0 μM, for dipropyl disulfide; from about 25.0 μM to about 250.0 μM, such as from about 90.0 μM to about 110.0 μM, for naringenin; and from about 10.0 μM to about 75.0 μM, such as from about 25.0 μM to about 35.0 μM, for α-hexylcinnamaldehyde.

25. The composition of claims 15-17, wherein the amount of at least one other compound in the composition is about 0.5 ppm, such as about 5 ppm, for garlic oil; about 0.5 ppm to about 50 ppm, such as about 15 ppm to about 25 ppm, for thyme red oil and lemongrass oil; about 0.1 ppm to about 10 ppm, such as about 0.75 ppm to about 2.5 ppm, for pyromellitic acid diimide.

26. An animal feed comprising the composition of any one of claims 1-25, for example, wherein the animal feed is an animal feed supplement, or an animal feed additive, or a supplementary feed.

27. Use of the composition according to claims 1 to 25 or the animal feed according to claim 16 for: a. Reducing methane emissions from animals; and / or b. inhibiting the production of methane by one or more methanogens in an animal; and / or c. Increase milk production and / or weight gain in animals.

28. The use according to claim 27, wherein the composition is consumed by an animal.

29. Use according to claim 27 or 28, wherein the animal is a ruminant, such as cattle, goats or sheep.

30. A method comprising: a. Reduce methane emissions from animals, and / or b. inhibiting the production of methane by one or more methanogens in an animal, and / or c. Increase milk production and / or weight gain in animals: The method comprises administering to the animal a composition according to any one of claims 1-25 or an animal feed according to claim 26.

31. The method of claim 30, wherein the animal is a ruminant such as a cow, goat or sheep.

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