Haloform reaction precursors as Anti-methanogenic formulations for administration to ruminants

Abstract

The present disclosure relates to compounds and compositions, for example, anti-methanogenic compositions, comprising a trihalomethyl ketone and methods of administering the anti-methanogenic composition to a ruminant to reduce methane emissions. Systems and associated methods of reducing methane are further provided.

Description

[0001] HALOFORM REACTION PRECURSORS AS ANTI-METHANOGENIC

[0002] FORMULATIONS FOR ADMINISTRATION TO RUMINANTS

[0003] CROSS REFERENCE TO RELATED APPLICATIONS

[0004] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 571,943, filed on March 29, 2024; and U.S. Provisional Patent Application No. 63 / 704,501, filed on October 7, 2024; the contents of each of which are incorporated herein by reference in their entireties.

[0005] TECHNICAL FIELD

[0006] The present disclosure relates to the use of stable intermediates of the haloform reaction as anti-methanogenic agents in ruminant livestock. Haloforms are very volatile, particularly in aqueous solution, and as such practical and accurate dosing is difficult. However, these precursors are stable in air with negligible volatility and will readily undergo hydrolysis (or amino lysis / alcoholysis) in the rumen to form the active haloform species in situ. This haloform produces an anti-methanogenic effect by inhibiting microbial production of methane. Furthermore, these precursor compounds being polyhalogenated compounds may also inhibit methane production.

[0007] BACKGROUND

[0008] In the realm of livestock management, one of the significant challenges is the control of methane emissions. Ruminants, such as cows, sheep, and goats, are known to produce methane as a byproduct of their digestive process. This methane is released into the atmosphere, contributing to greenhouse gas emissions and global warming. Furthermore, the production of methane also represents a loss of energy for the animal as the energy contained in the methane could have been used for growth or milk production. A reduction in methane emissions from ruminant livestock has been hypothesized to also yield an improvement in productivity through re-direction of energy that would be otherwise consumed in the production of methane (see Patra AK, Environ Monit Assess. 2012 Apr; 184(4): 1929-52, the contents of which is incorporated by reference in its entirety). This hypothesis is predicated on the hydrogen gas that is produced when methane production is inhibited being consumed as an energy source (see Morgavi DP et al., Animal. 2023 Jul;17 Suppl 3:100830, the contents of which is incorporated by reference in its entirety). Experimental data have also shown that energy gains are inconsistent; furthermore, theoretical calculations show that any energy gain would be modest. For this reason, phenolic compounds such as gallic acid or phloroglucinol as hydrogen acceptors can be used simultaneously to increase acetate and volatile fatty acid production (see Huang R. et al., Animal, 2023, 17(5), pp.100788, the contents of which is incorporated by reference in its entirety).

[0009] Bromoform is the most potent of these haloforms and is being commercialized as a chemical agent to reduce methane emissions arising from ruminant livestock. Unfortunately, aqueous solutions of bromoform are unstable due to its unfavorable air / water partition coefficient leading to significant volatility which makes its effective dosing in this form in a commercial livestock setting very challenging.

[0010] Various approaches are being explored for their potential to stabilize bromoform and other haloforms in a manner suitable for administration to ruminants. Ideally, the formulation would be stable during manufacture and storage and furthermore allow for its ready release in the rumen when dosed in the animal.

[0011] Asparagopsis seaweed is a natural source of bromoform, and various formulations have been developed by others where the bromoform is stabilized through extraction into food oils. In food oils, the intermolecular interaction between bromoform and the organic liquid is much stronger (i.e., relative to bromoform / water intermolecular interactions) resulting in a substantial reduction in volatility. Waxes (e.g., carnauba wax) and high melting point fats (e.g., hydrogenated coconut oil) can also be used to stabilize bromoform. However, fats can often bypass the rumen resulting in poor delivery efficiency to the rumen thus reducing the efficacy of these anti-methanogenic agents. When comparing a solid formulation to a liquid formulation, it might be expected that the volatility would be reduced due to greater viscosity which would restrict diffusion to the surface as predicted by the Einstein-Stokes relationship where it can evaporate into the air. However, in both cases the volatility will be significantly impacted by the specific surface area of either the liquid or solid. When the carrier is dispersed into a feed, the surface area increases dramatically leading to concomitant increase in the volatilization rate. Due to this volatility, this strategy is difficult to practically implement at scale over the long term. Similarly, the release rate in the rumen will be impacted by the surface area.

[0012] As a result, it is desirable to develop a precursor compound (also known as a prodrug) that has negligible volatility, is chemically stable, and can readily convert under in situ conditions to form the desired haloform upon hydrolysis (or alcoholysis / amino lysis) of the precursor. The present disclosure addresses a need for stabilized forms of bromoform for use in reducing methane production in ruminants without negatively affecting their health or productivity through accurate dosing.

[0013] SUMMARY

[0014] In some aspects, the present disclosure relates to the synthesis of trihalomethyl ketones (which mimic an intermediate in the haloform reaction) as precursors which upon hydrolysis in the rumen under relevant pH conditions will form the respective trihalomethane active ingredient and a carboxylic acid.

[0015] In some aspects, the present disclosure relates to the reaction of trihalomethyl ketones with amines in situ to form their respective trihalomethane and an amide.

[0016] In some aspects, the present disclosure relates to the reaction of trihalomethyl ketones with alcohols in situ to form their respective trihalomethane and an ester.

[0017] In other aspects, the present disclosure relates to the immobilization of these trihalomethyl ketone compounds onto cellulose from plant and grass material as a delivery mechanism.

[0018] In other aspects, the present disclosure provides animal feed additives and nutritional preparations for an animal comprising the anti-methanogenic compositions described herein. Preferably the anti-methanogenic agent is a haloform, more preferably bromoform.

[0019] In other aspects, the present disclosure provides trihalomethyl ketones as potential anti- methanogenic agents.

[0020] In still other aspects, the disclosure provides methods of reducing methane emission by administering a feed additive or nutritional preparation described herein to ruminants, such as cows, sheep, and goats.

[0021] In some embodiments, a compound or composition is provided, for example, an anti- methanogenic compound or composition, comprising, consisting essentially of, or consisting of: a trihalomethyl ketone having a structure of Formula (I): wherein R1represents hydrogen, substituted or unsubstituted Ci-Ce alkyl such methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl, neopentyl, isopentyl, pentan-2-yl, 3-pentyl, sec-isopentyl, 2-methylbutyl, hexyl, isohexyl, neohexyl, terthexyl, hexan-2-yl, hexan-3-yl, 2-methylpentyl, 3 -methylpentyl, 2,3-dimethylbutyl, substituted or unsubstituted Cs-C'x cycloalkyl or Ci-Cs-alkyl-Cs-Cs-cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopropyl-methyl, cyclobutylmethyl, cyclopentyl-methyl, cyclohexyl-methyl, cycloheptyl-methyl, cyclooctyl-methyl, cyclopropyl-ethyl, cyclobutyl-ethyl, cyclopentyl-ethyl, cyclohexyl-ethyl, cycloheptyl-ethyl, cyclooctyl-ethyl, cyclopropyl-propyl, cyclobutyl-propyl, cyclopentyl-propyl, cyclohexylpropyl, cycloheptyl-propyl, cyclooctyl-propyl, substituted or unsubstituted C5-C14 aryl or Ci- C3-alkyl-C5-Ci4-aryl, or -(CO)NR3(CO)NR4R5;

[0022] R2represents halogen; and

[0023] R3, R4, and R5each independently represents hydrogen, halogen, substituted or substituted C1-C4 alkyl such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tertbutyl; and optionally an agriculturally acceptable carrier.

[0024] In some embodiments, wherein R2represents Cl or Br.

[0025] In some embodiments, wherein R2represents Br.

[0026] In some embodiments, wherein R1is substituted by one or more halogens, -OH, - COOH, -NO2, -OCH3, -CH3, -CN, or -NH2.

[0027] In some embodiments, wherein R1represents -(CO)NR3(CO)NR4R5.

[0028] In some embodiments, wherein the trihalomethyl ketone is Formula (1-1): (1-1).

[0029] In some embodiments, wherein the agriculturally acceptable carrier is a hydrophobic carrier.

[0030] In some embodiments, wherein the agriculturally acceptable carrier comprises one or more of a vegetable oil, an animal fat, and / or a microbial oil, and / or a mixture thereof.

[0031] In some embodiments, wherein the vegetable oil comprises one or more of coconut oil, corn oil, cottonseed oil, olive oil, palm oil, peanut oil, canola oil, safflower oil, sesame oil, soybean oil, sunflower oil, almond oil, beech nut oil, Brazil nut oil, cashew oil, Jamaican cobnut oil, hazelnut oil, macadamia oil, mongongo nut oil, pecan oil, pine nut oil, pistachio oil, walnut oil, pumpkin seed oil, grapefruit seed oil, lemon oil, orange oil, avocado oil, poppyseed oil, tea seed oil, and / or apricot oil, and / or a mixture thereof.

[0032] In some embodiments, wherein the animal fat comprises one or more of fish oil, pig fat, chicken fat, cow fat, and / or butter, and / or a mixture thereof.

[0033] In some embodiments, wherein the microbial oil comprises oils produced by one or more of Yarrowia lipoytica, Schizochytrium sp., Crypthecodinium cohnii, Mortierella alpina, Mortierella elongate, Mortierella isabelline, Porphyridium cruentum, Mucor circinelloides , Aspergillus niger, Borago officinalis, Nitzschi laevis, Navicula pelliculosa, Cylindrotheca fusiformis, Scendesmus obtusiusculus, and / or Haema-tococcus pluvialis, and / or a mixture thereof.

[0034] In some embodiments, wherein the trihalomethyl ketone is amphiphilic.

[0035] In some embodiments, wherein the trihalomethyl ketone has a solubility to water (mwater / mFomuia (i)) of from about 0.1 to about 100, from about 0.5 to about 100, from about 1 to about 100, from about 1.25 to about 50, from about 1.5 to about 40, from about 1.75 to about 30, from about 2 to about 20, from about 2 to about 10, or from about 5 to about 10.

[0036] In some embodiments, wherein the trihalomethyl ketone has a solubility to food oil (rnWater / moii) of from about 0.1 to about 100, from about 0.5 to about 100, from about 1 to about 100, from about 1.25 to about 50, from about 1.5 to about 40, from about 1.75 to about 30, from about 2 to about 20, from about 2 to about 10, or from about 5 to about 10.

[0037] In some embodiments, wherein the trihalomethyl ketone has a viscosity under standard atmospheric conditions (25°C and pressure of 1 bar) of about 0.250 mPa-s to about 1000 mPa-s, about 0.500 mPa-s to about 1000 mPa-s, about 0.750 mPa-s to about 1000 mPa-s, about 1 mPa-s to about 1000 mPa-s, about 2.5 mPa-s to about 1000 mPa-s, about 5 mPa-s to about 1000 mPa-s, about 7.5 mPa-s to about 1000 mPa-s, about 10 mPa-s to about 1000 mPa-s, about 15 mPa-s to about 1000 mPa-s, about 20 mPa-s to about 1000 mPa-s, about 25 mPa-s to about 1000 mPa-s, about 25 mPa-s to about 750 mPa-s, about 25 mPa-s to about 500 mPa-s, about 25 mPa-s to about 250 mPa-s, about 25 mPa-s to about 100 mPa-s, about 25 mPa-s to about 75 mPa-s, or about 25 mPa-s to about 50 mPa-s.

[0038] In some embodiments, the anti-methanogenic composition further comprises a bulking agent, wherein the bulking agent may comprise polysaccharides in aspects. In some embodiments, the polysaccharides comprise cellulose, hemicellulose, starch, inulin, chitin, and / or pectin, and / or a mixture thereof.

[0039] In some embodiments, wherein: cellulose is derived from one or more of grass (such as Bermuda grass, brome, oatgrass, fescue, false oat grass, meadow grasses, orchard grass, ryegrass, and timothy-grass), millet, oats, sorghum, soybean, cotton, hemp, flax, jute, corn (such as corn cob, maize straw), banana, wheat (such as wheat straw), cork bark, curaua, kenaf, nettle, ramie, rice husk, hardwood, softwood, sugar can bagasse, sisal, regenerated celluloses (such as viscose and cellophane), and / or bacterial cellulose (produced by species such as Acetobacter pasteurianum, Acetobacter xylinum, Acetobacter rancens, Komagataeibacter xylinus, Sarcina ventriculi, and / or Bacterium xylinoides , hemicellulose is derived from one or more of grass (such as Bermuda grass, brome, oatgrass, fescue, false oat grass, meadow grasses, orchard grass, ryegrass, and timothy-grass), millet, oats, sorghum, soybean, cotton, hemp, flax, jute, corn (such as corn cob, maize straw), banana, wheat (such as wheat straw), cork bark, curaua, kenaf, nettle, ramie, rice husk, hardwood, softwood, sugar can bagasse, and / or sisal; starch is derived from one or more of rice, wheat, maize, potato, cassava, acorn, arrowroot, arracacha, banana, barley, breadfruit, buckwheat, canna, colocasia, cuckoo-pint, katakuri, kudzu, malanga, millet, oat, oca, Polynesian arrowroot, sago, sorghum, sweet potatoe, rye, taro, chestnut, water chestnut, yam, sorghum, soybean, fava, lentil, mung bean, adzuki bean, pea, and / or chickpea; inulin is derived from one or more of Agave, banana, burdock, camas, chicory, coneflower, costus, dandelion, elecampane, garlic, globe artichoke, Jerusalem artichoke, jicama, leopard's bane, mugwort, onion, yam, and / or daisy; chitin is derived from fungi and / or arthropods; pectin is derived from one or more of pear, apple, guava, quince, plum, citrus (such as orange, lemon, grapefruit, pomelo, lime), apricot, cherry, carrot, and / or rose hip.

[0040] In some embodiments, the polysaccharides are functionalized with a trihalomethyl ketone through an ether linkage, an amine linkage, a thioether linkage, an ester linkage, a thioester linkage, an amide linkage, a thioamide linkage, or a thionoester linkage.

[0041] In some embodiments, wherein the bulking agent may comprise one or more high surface area materials.

[0042] In some embodiments, wherein the one or more high surface area materials have a surface area of 50-5000 m2 / g, optionally 100-5000 m2 / g, optionally 200-5000 m2 / g, optionally 300-5000 m2 / g, optionally 500-5000 m2 / g, optionally 500-4000 m2 / g, optionally 500-3000 m2 / g, optionally 750-3000 m2 / g, optionally 900-3000 m2 / g, optionally 1000-3000 m2 / g, optionally 500-1500 m2 / g.

[0043] In some embodiments, wherein the one or more high surface area materials comprise activated carbons and / or crosslinked polymers, and / or a mixture thereof, wherein the activated carbons comprise: powdered activated carbon (PAC), granular activated carbon (GAC), extruded activated carbon (EAC), bead activated carbon (BAC), impregnated carbon, polymer coated carbon, woven carbon, carbon nanotubes (CNTs), graphene, carbon aerogels, carbon black, carbon fibers, carbon nanofibers (CNFs), carbon cloth, carbon paper, and / or carbon foam, wherein the crosslinked polymers comprise: polydimethylsiloxane (PDMS) and silica- based polymers, polyacrylate resins, graphene oxide and modified graphene materials, poly(N- vinyl-2 -pyrrolidone) (PVP) and PVP -based materials, styrene-divinylbenzene (Styrene-DVB) copolymers, amberchrom resins, chromabond resins, XAD resins, polystyrene, polyacrylic, polyacrylic ester, and / or cation exchange resin.

[0044] In some embodiments, wherein one or more aromatic rings of the one or more high surface area materials are modified to add one or more tribromomethylketone functional groups.

[0045] In some embodiments, wherein the trihalomethyl ketone has a partition coefficient (log P), as defined under an octanol-water system, of from about -2.0 to about 2.5, from about -1.75 to about 2.5, from about -1.5 to about 2.5, from about -1.25 to about 2.5, from about -1 to about 2.5, from about -1 to about 2.25, from about -1 to about 2, from about -1 to about 1.75, from about -1 to about 1.5, from about -1 to about 1.25, from about -1 to about 1, from about -0.75 to about 1, from about -0.5 to about 1, from about -0.5 to about 0.75, or from about -0.5 to about 0.5.

[0046] In aspects, cellulose compounds described herein are used to deliver compounds or compositions described herein to an animal in need thereof.

[0047] In some embodiments, an animal feed comprising the anti-methanogenic composition is provided.

[0048] In some embodiments, wherein the animal feed is in the form of a powder, granule, pellet, feed block, lick block, gel, ointment, cream, paste, solution, suspension, or emulsion.

[0049] In some embodiments, wherein the animal feed further comprising a roughage, cereal, starch, vegetable waste, vitamin, mineral, trace element, emulsifier, aromatizing product, binder, colorant, odorant, thickening agent, or a combination thereof. In some embodiments, wherein the animal feed comprises from about 0.0001 wt.% to about 10 wt.%, from about 0.001 wt.% to about 9.5 wt.%, from about 0.005 wt.% to about 9 wt.%, from about 0.01 wt.% to about 8.5 wt.%, from about 0.05 wt.% to about 8 wt.%, from about 0.1 wt.% to about 7.5 wt.%, from about 0.9 wt.% to about 7 wt.%, from about 1 wt.% to about 6 wt.%, from about 1.5 wt.% to about 5.5 wt.%, from about 2 wt.% to about 5 wt.%, from about 2.5 wt.% to about 4.5 wt.%, or from about 3 wt.% to about 4 wt.% the anti- methanogenic composition based on the total dry weight of the animal feed.

[0050] In some embodiments, wherein the animal feed comprises from about 0.4 wt.% to about 9.5 wt.%, from about 0.5 wt.% to about 9 wt.%, from about 0.6 wt.% to about 8.5 wt.%, from about 0.7 wt.% to about 8 wt.%, from about 0.8 wt.% to about 7.5 wt.%, from about 0.9 wt.% to about 7 wt.%, from about 1 wt.% to about 6 wt.%, from about 1.5 wt.% to about 5.5 wt.%, from about 2 wt.% to about 5 wt.%, from about 2.5 wt.% to about 4.5 wt.%, or from about 3 wt.% to about 4 wt.% the anti-methanogenic composition based on the total dry weight of the animal feed.

[0051] In some embodiments, a method for reducing methane emissions from a ruminant is provided, comprising, consisting essentially of, or consisting of: administering to the ruminant a composition comprising a trihalomethyl ketone having a structure of Formula (I): wherein

[0052] R1represents hydrogen, substituted or unsubstituted Ci-Ce alkyl such methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl, neopentyl, isopentyl, pentan-2-yl, 3-pentyl, sec-isopentyl, 2-methylbutyl, hexyl, isohexyl, neohexyl, terthexyl, hexan-2-yl, hexan-3-yl, 2-methylpentyl, 3 -methylpentyl, 2,3-dimethylbutyl, substituted or unsubstituted Cs-C'x cycloalkyl or Ci-Cs-alkyl-Cs-Cs-cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopropyl-methyl, cyclobutylmethyl, cyclopentyl-methyl, cyclohexyl-methyl, cycloheptyl-methyl, cyclooctyl-methyl, cyclopropyl-ethyl, cyclobutyl-ethyl, cyclopentyl-ethyl, cyclohexyl-ethyl, cycloheptyl-ethyl, cyclooctyl-ethyl, cyclopropyl-propyl, cyclobutyl-propyl, cyclopentyl-propyl, cyclohexyl- propyl, cycloheptyl-propyl, cyclooctyl-propyl, substituted or unsubstituted C5-C14 aryl or Ci- C3-alkyl-C5-Ci4-aryl, or -(CO)NR3(CO)NR4R5;

[0053] R2represents halogen; and

[0054] R3, R4, and R5each independently represents hydrogen, halogen, substituted or substituted C1-C4 alkyl such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, .sec-butyl, tertbutyl; and optionally an agriculturally acceptable carrier.

[0055] In some embodiments, the trihalomethyl ketone of the composition to be administered to the ruminant is Formula (1-1):

[0056] (1-1).

[0057] In some embodiments, wherein the trihalomethyl ketone is administered in an amount of between 1 mg / kg and 1000 mg / kg dry matter intake (DMI) to the ruminant, between 1 mg / kg and 750 mg / kg DMI to the ruminant, between 1 mg / kg and 500 mg / kg DMI to the ruminant, between 1 mg / kg and 400 mg / kg DMI to the ruminant, between 1 mg / kg and 300 mg / kg DMI to the ruminant, between 1 mg / kg and 200 mg / kg DMI to the ruminant, between 1 mg / kg and 100 mg / kg DMI to the ruminant, between 5 mg / kg and 100 mg / kg DMI to the ruminant, between 10 mg / kg and 100 mg / kg DMI to the ruminant, or between 25 mg / kg and 75 mg / kg DMI to the ruminant.

[0058] In some embodiments, wherein the ruminant is a member of the family Bovidae.

[0059] In some embodiments, wherein the ruminant is Bos taurus or Bos indicus.

[0060] In some embodiments, wherein the ruminant is Ovis aries or Capra hircus.

[0061] In some embodiments, wherein the trihalomethyl ketone is administered orally, parenterally, or by injection. In one embodiment, the trihalomethyl ketone is administered orally.

[0062] In some embodiments, wherein the trihalomethyl ketone is administered in an animal feed.

[0063] In some embodiments, wherein the trihalomethyl ketone is administered by a feed dispenser. In other embodiments, the methane emissions are reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% compared to an untreated control ruminant.

[0064] In an aspect, compounds and compositions described herein, for example those of Formula I and 1-1, are used in a ruminant and wherein the ruminant is of the family Bovidae. In another aspect, compounds and compositions described herein, for example those of F ormula I and I- 1 , are used in a ruminant and wherein the ruminant is Bos taurus or Bos indicus . In yet another aspect, compounds and compositions described herein, for example those of F ormula I and I- 1 , are used in a ruminant and wherein the ruminant is Bos taurus or Bos indicus .

[0065] In a non-limiting aspect, compounds and compositions described herein, for example those of Formula I and 1-1, are used in a ruminant and wherein the ruminant is of the family Bovidae and wherein the compound or composition is not used to treat a human. In another aspect, compounds and compositions described herein, for example those of Formula I and I- 1 , are used in a ruminant and wherein the ruminant is Bos taurus or Bos indicus and wherein the compound or composition is not used to treat a human. In yet another aspect, compounds and compositions described herein, for example those of Formula I and 1-1, are used in a ruminant and wherein the ruminant is Bos taurus or Bos indicus and wherein the compound or composition is not used to treat a human.

[0066] In an aspect, compounds and compositions described herein, for example those of Formula I and 1-1, are used in cattle. In a non-limiting aspect described herein, compounds and compositions described herein, for example those of F ormula I and I- 1 , are not used as a human therapy or treatment. In another non-limiting aspect, compounds and compositions described herein, for example those of Formula I and 1-1, are used as a prodrug in cattle and not in humans. In another non-limiting aspect, compounds and compositions described herein, for example those of Formula I and 1-1, are not suitable for use in humans.

[0067] The following description is intended only by way of example, and simply illustrates certain selected embodiments of devices, systems, and processes that are consistent with the disclosed subject matter as claimed herein.

[0068] BRIEF DESCRIPTION OF THE DRAWINGS

[0069] FIG. 1 depicts a general reaction where a tribromomethyl ketone is hydrolyzed to generate bromoform.

[0070] FIG. 2 depicts a synthetic scheme for preparing tribromopyruvyl urea. FIG. 3 depicts general reactions where a tribromomethyl ketone reacts with a base or an amine to form bromoform.

[0071] FIG. 4 depicts a synthetic scheme for preparing a tribromomethyl ketone via oxidation of the corresponding tribromomethyl alcohol. PCC: pyridinium chlorochromate; CH2Q2: dichloromethane; IBX: 2-iodoxybenzoic acid.

[0072] FIG. 5 depicts a synthetic scheme for preparing a tribromomethyl ketone from an alkynoic acid. AgOAc: silver acetate.

[0073] DETAILED DESCRIPTION

[0074] Preferred features, embodiments and variations of the invention may be discerned from the following detailed description which provides sufficient information for those skilled in the art to perform the invention. The detailed description is not to be regarded as limiting the scope of the preceding summary of the invention in any way.

[0075] In compliance with the statute, the invention has been described in a language more or less specific to structural or methodical features. The term “comprises” and its variations, such as “comprising” and “comprised of’ is used throughout in an inclusive sense and not to the exclusion of any additional features. It is to be understood that the invention is not limited to specific features shown or described since the means herein described comprises preferred forms of putting the invention into effect. The invention is, therefore, claimed in any of its forms or modifications within the proper scope of the appended claims appropriately interpreted by those skilled in the art.

[0076] Throughout the specification and claims (if present), unless the context requires otherwise, the term “substantially” or “about” will be understood to not be limited to the value for the range qualified by the terms.

[0077] It is to be understood that unless specifically stated otherwise, references to “a,” “an,” and / or “the” may include one or more than one and that reference to an item in the singular may also include the item in the plural. Reference to an element by the indefinite article “a,” “an” and / or “the” does not exclude the possibility that more than one of the elements are present, unless the context clearly requires that there is one and only one of the elements. As used herein, the term “comprise,” and conjugations or any other variation thereof, are used in its non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded.

[0078] A “ruminant” is a mammal of the order Artiodactyla that digests plant-based food by initially softening and partially fermenting it within the animal's first stomach chambers, then regurgitating the semi-digested mass, now known as cud, and chewing it again. The process of rechewing the cud to further break down plant matter and stimulate digestion is called “ruminating”. Ruminants have a digestive tract with four chambers, namely the rumen, reticulum, omasum and abomasum. In the first two chambers, the rumen and the reticulum, the food is mixed with saliva and separates into layers of solid and liquid material. Solids clump together to form the cud, or bolus. The cud is then regurgitated, chewed slowly to completely mix it with saliva, which further breaks down fibers. Fiber, especially cellulose, is broken down into glucose in these chambers by symbiotic anaerobic bacteria, protozoa and fungi. The broken-down fiber, which is now in the liquid part of the contents, then passes through the rumen into the next stomach chamber, the omasum. The food in the abomasum is digested much like it would be in the monogastric stomach. Digested gut contents are finally sent to the small intestine, where the absorption of the nutrients occurs. Almost all the glucose produced by the breaking down of cellulose is used by the symbiotic bacteria. Ruminants get their energy from the volatile short chain fatty acids (VFAs) produced by the bacteria, namely acetate, propionate, butyrate, valerate, and isovalerate. Ruminants included cattle, goats, sheep, giraffes, yaks, deer, antelope, and others.

[0079] As used herein (if present), the term “bovid” includes any member of the family Bovidae, which include hoofed mammals such as antelope, sheep, goats, and cattle, among others.

[0080] As used herein, the term “reducing” includes the reduction of amount of substance in comparison with a reference. For example, the reduction in the amount of total gas and / or methane produced by a ruminant animal or animals administered a composition according to the present invention, relative to an animal or animals not administered a composition of the present invention. The reduction can be measured in vitro with an artificial rumen system that simulates anaerobic fermentation, breath measurement over 24 hrs (e.g., SFB cannisters mounted on individual animals or spot samples intermittently using sensors operating via analysis of breath (e.g., green feed units), with micrometeorological methods (most commonly in field), in vivo with animals confined in respiration chambers, or with other methods known in the art. It is within the knowledge and skill of those trained in the art to assess enteric methanogenesis by a ruminant animal.

[0081] As used herein, the term “reducing methane production” refers to the reduction of methane produced in the gastro-intestinal tract. The term includes the specific volume of methane generated as a result of anaerobic fermentation, for example, in the systems described herein. Fermentation in the rumen and the gut of a ruminant gives rise to production of methane. The present invention aims to reduce this process, such as to reduce the total amount of methane produced in the gastro-intestinal tract. It is within the knowledge and skill of those trained in the art to assess methane production by a ruminant animal.

[0082] The term “methanogenic” refers to the process of producing methane by a microbially generated pathway. An anti-methanogenic composition is one that interferes with the methanogenic pathway so as to reduce the amount of methane produced.

[0083] As used herein, the term “tribromopyruvyl urea” refers to the compound having the IUPAC name of 3,3,3-tribromo-A-carbamoyl-2-oxopropanamide and the chemical structure shown below.

[0084] As used herein, “alkyl” denotes a straight-chain or branched open-chain, saturated hydrocarbon radical which is optionally mono- or polysubstituted, and in the latter case is referred to as “substituted alkyl”.

[0085] Exemplary and non-limiting alkyls include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl, neopentyl, isopentyl, pentan-2-yl, 3-pentyl, sec-isopentyl, 2-methylbutyl, hexyl, isohexyl, neohexyl, tert-hexyl, hexan-2-yl, hexan-3-yl, 2- methylpentyl, 3 -methylpentyl, and 2, 3 -dimethylbutyl.

[0086] As described herein, “cycloalkyl” denotes a cyclic alkyl group, saturated hydrocarbon radical which is optionally mono- or polysubstituted, and in the latter case is referred to as “substituted cycloalkyl”. Exemplary alkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0087] As described herein, “aryl” denotes a functional group derived from an aromatic ring, which is optionally mono- or polysubstituted, and in the latter case is referred to as “substituted aryl”. Exemplary aryls include phenyl, tolyl, xylyl, and naphthyl.

[0088] Preferred substituents are alkoxy, cyano, alkylthio, amino or nitro groups, particular preference being given to methoxy, methyl, cyano, or nitro.

[0089] The term “halogen” denotes, for example, fluorine, chlorine, bromine or iodine. If the term is used for a radical, "halogen" denotes, for example, a fluorine, chlorine, bromine or iodine atom. Synthesis of Stable Haloform Reaction Precursors

[0090] The haloform reaction is predicated on sequential bromination reactions of typically a methyl ketone or 1,3 -diketone followed by hydrolysis which typically yields a haloform and carboxylic acids or carbon dioxide. By developing synthetic pathways where the pH and temperature conditions for bromination and hydrolysis are significantly different, it is possible to readily isolate the halogenated precursors. These precursors can then convert into the active haloform compound in the rumen. Alternatively, other synthetic pathways to these trihalomethyl ketone species can be utilized (see Gulizhabaier, A., and A. A. Rexit, Russian Journal of Organic Chemistry 57.5 (2021): 809-815; Jayaraman, Aravindan, et al., Advanced Synthesis & Catalysis 360.20 (2018): 3978-3989; and Cavallito, Chester J., and Clayton S. Smith, Journal of the American Chemical Society 63.4 (1941): 995-998), the contents of each of these references is hereby incorporated by reference in their entirety. Another strategy is to directly react bromoform with an aldehyde to yield a tribromomethyl carbinol compound (see Hasimujiang, Balati, et al. ChemistrySelect 4.19 (2019): 5560-5562, the contents of which is incorporated by reference in its entirety) which can then be oxidized to yield the desired tribromomethyl ketone (see Hasimujiang, Balati, et al. Synthetic Communications 48.8 (2018): 887-891, the content of which are herein incorporated by reference in their entireties). These brominated ketone species will generally have a much lower volatility compared to their corresponding haloform and can then be ingested by the ruminant where it will convert to the respective haloform in the rumen through a hydrolysis reaction. Furthermore, these compounds may also be activated to release bromoform in the presence of amines such as urea (see Dohi, Souya, Katsuhiko Moriyama, and Hideo Togo. European Journal of Organic Chemistry 2013.34 (2013): 7815-7822, the contents of which is herein incorporated by reference in its entirety).

[0091] The trihalomethyl ketone may be hydrolysable under pH conditions from 6.2 to 7.0 which are typical of values found in the rumen of ruminants. On high grass diets, particularly where they are highly fibrous, the range is generally 6.8 - 7.2. For animals consuming diets high in grain (e.g., feedlot) or containing significant amounts of grains or other fermentable material (e.g., dairy), the pH of the rumen is inevitably slightly more acidic (e.g., 6.5-7).

[0092] The trihalomethyl ketone may be a trichloromethyl ketone, a tribromomethyl ketone or a triiodomethyl ketone, or combination thereof. Trihalomethyl Ketone

[0093] The trihalomethyl ketone may have a structure of Formula (I): wherein

[0094] R1represents hydrogen, substituted or unsubstituted Ci-Ce alkyl such methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl, neopentyl, isopentyl, pentan-2-yl, 3-pentyl, sec-isopentyl, 2-methylbutyl, hexyl, isohexyl, neohexyl, terthexyl, hexan-2-yl, hexan-3-yl, 2-methylpentyl, 3 -methylpentyl, 2,3-dimethylbutyl, substituted or unsubstituted Cs-C'x cycloalkyl or Ci-Cs-alkyl-Cs-Cs-cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopropyl-methyl, cyclobutylmethyl, cyclopentyl-methyl, cyclohexyl-methyl, cycloheptyl-methyl, cyclooctyl-methyl, cyclopropyl-ethyl, cyclobutyl-ethyl, cyclopentyl-ethyl, cyclohexyl-ethyl, cycloheptyl-ethyl, cyclooctyl-ethyl, cyclopropyl-propyl, cyclobutyl-propyl, cyclopentyl-propyl, cyclohexylpropyl, cycloheptyl-propyl, cyclooctyl-propyl, substituted or unsubstituted C5-C14 aryl or Ci- C3-alkyl-C5-Ci4-aryl, or -(CO)NR3(CO)NR4R5;

[0095] R2represents halogen; and

[0096] R3, R4, and R5each independently represents hydrogen, halogen, substituted or substituted C1-C4 alkyl such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tertbutyl.

[0097] In some embodiments, wherein R2represents Cl or Br.

[0098] In some embodiments, wherein R2represents Br.

[0099] In some embodiments, wherein R1is substituted by one or more halogens, -OH, - COOH, -NO2, -OCH3, -CH3, -CN, or -NH2.

[0100] In some embodiments, wherein R1represents -(CO)NR3(CO)NR4R5.

[0101] In some embodiments, wherein the trihalomethyl ketone is Formula (1-1):

[0102] In some embodiments, the trihalomethyl ketone is l-(trichloroacetyl)urea (C3H3CI3N2O2), trichloroacetyl chloride (CChCOCl), tribromoacetyl chloride (CB COCl), tribromoacetone (CBr3COCH3), trichloroacetone (CC13COCH3), 2,2,2-trichloro-l- (trichloromethyl)ethenone, trichloroacetamide (CCLCONH2).

[0103] The trihalomethyl ketone of Formula (I) may be amphiphilic, i.e., may be soluble in water and in hydrophobic solvents.

[0104] The solubility of a solute in a solvent may be defined as: msv / msu, in which msvrepresents the mass of the solvent, and msurepresents the mass of the solute. The trihalomethyl ketone of Formula (I) may have a solubility to water (mWater / mFomuia (i)) of from about 0.1 to about 100, from about 0.5 to about 100, from about 1 to about 100, from about 1.25 to about 50, from about 1.5 to about 40, from about 1.75 to about 30, from about 2 to about 20, from about 2 to about 10, or from about 5 to about 10. In addition, the trihalomethyl ketone of Formula (I) may have a solubility to food oil (mWater / moii) of from about 0.1 to about 100, from about 0.5 to about 100, from about 1 to about 100, from about 1.25 to about 50, from about 1.5 to about 40, from about 1.75 to about 30, from about 2 to about 20, from about 2 to about 10, or from about 5 to about 10.

[0105] Viscosity (dynamic viscosity) is defined as (mass / length) / time. The viscosity (dynamic viscosity) of a fluid may be expressed as, under standard atmospheric conditions (25°C and pressure of 1 bar), the millipascal second (mPa-s) value of the fluid. The trihalomethyl ketone of Formula (I) may have a viscosity under standard atmospheric conditions (25°C and pressure of 1 bar) of about 0.250 mPa-s to about 1000 mPa-s, about 0.500 mPa-s to about 1000 mPa-s, about 0.750 mPa-s to about 1000 mPa-s, about 1 mPa-s to about 1000 mPa-s, about 2.5 mPa-s to about 1000 mPa-s, about 5 mPa-s to about 1000 mPa-s, about 7.5 mPa-s to about 1000 mPa-s, about 10 mPa-s to about 1000 mPa-s, about 15 mPa-s to about 1000 mPa-s, about 20 mPa-s to about 1000 mPa-s, about 25 mPa-s to about 1000 mPa-s, about 25 mPa-s to about 750 mPa-s, about 25 mPa-s to about 500 mPa-s, about 25 mPa-s to about 250 mPa-s, about 25 mPa-s to about 100 mPa-s, about 25 mPa-s to about 75 mPa-s, or about 25 mPa-s to about 50 mPa-s. Partition coefficient is the ratio of the concentration of a substance in one medium or phase to the concentration in a second phase when the two concentrations are at equilibrium; the logarithm of the ratio is expressed as log P. The octanol-water system is a solvent system commonly used for evaluating solute properties in the chemical and pharmaceutical sciences.

[0106] The trihalomethyl ketone of Formula (I) may have a partition coefficient (log P), as defined under an octanol-water system, of from about -2.0 to about 2.5, from about -1.75 to about 2.5, from about -1.5 to about 2.5, from about -1.25 to about 2.5, from about -1 to about 2.5, from about -1 to about 2.25, from about -1 to about 2, from about -1 to about 1.75, from about -1 to about 1.5, from about -1 to about 1.25, from about -1 to about 1, from about -0.75 to about 1, from about -0.5 to about 1, from about -0.5 to about 0.75, or from about -0.5 to about 0.5.

[0107] Anti-Methanogenic Composition

[0108] Also provided herein is a composition, for example an anti-methanogenic composition, comprising, consisting essentially of, or consisting of a trihalomethyl ketone of Formula (I) described herein.

[0109] Agriculturally Acceptable Carrier

[0110] In addition to the trihalomethyl ketone of Formula (I), the composition, for example an anti-methanogenic composition, may further comprise an agriculturally acceptable carrier. In some embodiments, the agriculturally acceptable carrier is a hydrophobic carrier. In certain aspects, the trihalomethyl ketone of Formula (I) is dissolved in a hydrophobic carrier (e.g., a food oil) prior to being added to the water to prevent hydrolysis prior to reaching the rumen.

[0111] The hydrophobic carrier may be a vegetable oil, an animal fat, and / or a microbial oil, and / or a mixture thereof. Vegetable oil includes, but not limited to, coconut oil, corn oil, cottonseed oil, olive oil, palm oil, peanut oil, canola oil, safflower oil, sesame oil, soybean oil, sunflower oil, almond oil, beech nut oil, Brazil nut oil, cashew oil, Jamaican cobnut oil, hazelnut oil, macadamia oil, mongongo nut oil, pecan oil, pine nut oil, pistachio oil, walnut oil, pumpkin seed oil, grapefruit seed oil, lemon oil, orange oil, avocado oil, poppyseed oil, tea seed oil, and / or apricot oil, and / or a mixture thereof. Animal fat includes, but not limited to, fish oil, pig fat, chicken fat, cow fat, and / or butter, and / or a mixture thereof. Microbial oil includes, but not limited to, oils produced by Yarrowia lipoytica, Schizochytrium sp., Crypthecodinium cohnii, Mortierella alpina, Mortierella elongate, Mortierella isabelline, Porphyridium cruentum, Mucor circinelloides , Aspergillus niger, Borago officinalis, Nitzschi laevis, Navicula pelliculosa, Cylindrotheca fusiformis, Scendesmus obtusiusculus, and / or Haema-tococcus pluvialis, and / or a mixture thereof.

[0112] Bulking Agent

[0113] In addition to the trihalomethyl ketone of Formula (I), the composition, for example an anti-methanogenic composition, may further comprise one or more bulking agents. The addition of bulking agents helps in immobilizing the trihalomethyl ketone of Formula (I), thereby promoting the delivery of the trihalomethyl ketone. The bulking agent may also act as an anticaking agent.

[0114] In some embodiments, the bulking agent comprises polysaccharides. In some embodiments, the bulking agent comprises one or more high surface area materials.

[0115] The polysaccharides may comprise cellulose, hemicellulose, starch, inulin, chitin, and / or pectin, and / or a mixture thereof. Details of each species of polysaccharides are further described below.

[0116] Cellulose

[0117] Cellulose is a polysaccharide consisting of a linear chain of glucose units. The chemical structure of cellulose may be expressed by the following general formula.

[0118] Cellulose may be derived from one or more of grass (such as Bermuda grass, brome, oatgrass, fescue, false oat grass, meadow grasses, orchard grass, ryegrass, and timothy-grass), millet, oats, sorghum, soybean, cotton, hemp, flax, jute, corn (such as corn cob, maize straw), banana, wheat (such as wheat straw), cork bark, curaua, kenaf, nettle, ramie, rice husk, hardwood, softwood, sugar can bagasse, sisal, regenerated celluloses (such as viscose and cellophane), and / or bacterial cellulose (produced by species such o Acetobacter pasteurianum, Acetobacter xylinum, Acetobacter rancens, Komagataeibacter xylinus, Sarcina ventriculi, and / or Bacterium xy lino ides'). Hemicellulose

[0119] Hemicelluloses are polysaccharides composed of diverse sugars such as glucose, mannose, galactose, rhamnose, xylose, and / or arabinose, as well as the acidified forms of these sugars. Common motifs of hemicelluloses include xylan, glucuronoxylan, arabinoxylan, glucomannan, and xyloglucan.

[0120] (Glucomannan) Hemicellulose may be derived from one or more of grass (such as Bermuda grass, brome, oatgrass, fescue, false oat grass, meadow grasses, orchard grass, ryegrass, and timothygrass), millet, oats, sorghum, soybean, cotton, hemp, flax, jute, com (such as corn cob, maize straw), banana, wheat (such as wheat straw), cork bark, curaua, kenaf, nettle, ramie, rice husk, hardwood, softwood, sugar can bagasse, and / or sisal.

[0121] Starch

[0122] Starch is a polysaccharide consisting of glucose units joined by glycosidic bonds. Starch includes the linear and helical amylose, and the branched amylopectin. General formulas of starch are shown below.

[0123] (Amylopectin) Starch may be derived from one or more of rice, wheat, maize, potato, cassava, acorn, arrowroot, arracacha, banana, barley, breadfruit, buckwheat, canna, colocasia, cuckoo-pint, katakuri, kudzu, malanga, millet, oat, oca, Polynesian arrowroot, sago, sorghum, sweet potatoe, rye, taro, chestnut, water chestnut, yam, sorghum, soybean, fava, lentil, mung bean, adzuki bean, pea, and / or chickpea.

[0124] Inulin

[0125] Inulins are polysaccharides and belong to a class of dietary fibers known as fructans. Inulins are comprised of chain-terminating glucosyl moieties and a repetitive fructosyl moiety. The degree of polymerization of inulins ranges from 2 to 60. The general formula of inulins may be expressed as follows.

[0126]

[0127] Inulin may be derived from one or more of Agave, banana, burdock, camas, chicory, coneflower, costus, dandelion, elecampane, garlic, globe artichoke, Jerusalem artichoke, jicama, leopard's bane, mugwort, onion, yam, and / or daisy.

[0128] Chitin

[0129] Chitin is a long-chain polymer of A-acetylglucosamine. The general formula of chitin may be expressed as follows. Chitin may be derived from fungi and / or arthropods.

[0130] Pectin

[0131] Pectins are polysaccharides rich in galacturonic acid and / or substituted galacturonic acid. Pectins may also comprise sugars such as rhamnose, galactose, arabinose, xylose, and / or apiose. Pectins typically have a molecular weight of about 60,000 to about 130,000 g / mol. The general formula of pectins may be expressed as follows.

[0132] Pectin may be derived from one or more of pear, apple, guava, quince, plum, citrus (such as orange, lemon, grapefruit, pomelo, lime), apricot, cherry, carrot, and / or rose hip.

[0133] Polysaccharides added to the composition as bulking agent may help in immobilizing the trihalomethyl ketone of Formula (I), thereby promoting the delivery of the trihalomethyl ketone. In some embodiments, the polysaccharides may chemically bond to the trihalomethyl ketone of Formula (I), thereby immobilizing these trihalomethyl ketone compounds. In some embodiments, the chemical bond linking the polysaccharides and the trihalomethyl ketone compounds may be, for example, an ester linkage, an amide linkage, an ether linkage, an amine linkage, a thioamide linkage, a carbamate linkage, a thioester linkage, a thionoester linkage, and / or a thioether linkage. Exemplary chemical linkages between the polysaccharides and the trihalomethyl ketone compounds are illustrated below.

[0134]

[0135] High surface area materials are materials with high surface area. Typically, high surface area materials have a surface area of at least 50 m2 / g, as measured by the BET method (Brunauer-Emmett-Teller adsorption method). The high surface area materials applied in the present invention typically have a surface area of 50-5000 m2 / g, optionally 100-5000 m2 / g, optionally 200-5000 m2 / g, optionally 300-5000 m2 / g, optionally 500-5000 m2 / g, optionally 500-4000 m2 / g, optionally 500-3000 m2 / g, optionally 750-3000 m2 / g, optionally 900-3000 m2 / g, optionally 1000-3000 m2 / g, optionally 500-1500 m2 / g. Details of each species of high surface area materials are further described below. Activated Carbon

[0136] Activated carbons are complex products which are difficult to classify on the basis of their behaviour, surface characteristics and other fundamental criteria. However, a broad classification is made for general purposes based on their size, preparation methods, and industrial applications.

[0137] Powdered Activated Carbon (PAC)

[0138] Normally, activated carbons (R 1) are made in particulate form as powders or fine granules less than 1.0 mm in size with an average diameter between 0.15 and 0.25 mm. Thus, they present a large surface to volume ratio with a small diffusion distance. Activated carbon (R 1) is defined as the activated carbon particles retained on a 50-mesh sieve (0.297 mm).

[0139] Powdered activated carbon (PAC) material is finer material. PAC is made up of crushed or ground carbon particles, 95-100% of which will pass through a designated mesh sieve. The ASTM classifies particles passing through an 80-mesh sieve (0.177 mm) and smaller as PAC. It is not common to use PAC in a dedicated vessel, due to the high head loss that would occur. Instead, PAC is generally added directly to other process units, such as raw water intakes, rapid mix basins, clarifiers, and gravity filters.

[0140] Granular Activated Carbon (GAC)

[0141] Granular activated carbon (GAC) has a relatively larger particle size compared to powdered activated carbon and consequently presents a smaller external surface. Diffusion of the adsorbate is thus an important factor. Granular activated carbon is suitable for adsorption of gases and vapours because gaseous substances diffuse rapidly. Granulated activated carbons are used for air filtration and water treatment, as well as for general deodorization and separation of components in flow systems and in rapid mix basins. GAC can be obtained in either granular or extruded form. GAC is designated by sizes such as 8x20, 12x40, 20x40, or 8x30 for liquid phase applications and 4x6, 4x8 or 4x 10 for vapor phase applications. A 20x40 carbon is made of particles that will pass through a U.S. Standard Mesh Size No. 20 sieve (0.84 mm) (generally specified as 85% passing) but be retained on a U.S. Standard Mesh Size No. 40 sieve (0.42 mm) (generally specified as 95% retained). AWWA (1992) B604 uses the 50- mesh sieve (0.297 mm) as the minimum GAC size. The most popular aqueous-phase carbons are the 12x40 and 8x30 sizes because they have a good balance of size, surface area, and head loss characteristics. Extruded Activated Carbon (EAC)

[0142] Extruded activated carbon (EAC) combines powdered activated carbon with a binder, which are fused together and extruded into a cylindrical shaped activated carbon block with diameters from 0.8 to 130 mm. These are mainly used for gas phase applications because of their low pressure drop, high mechanical strength and low dust content.

[0143] Bead Activated Carbon (BAC)

[0144] Bead activated carbon (BAC) is made from petroleum pitch and supplied in diameters from approximately 0.35 to 0.80 mm. Like EAC, it is also noted for its low pressure drop, high mechanical strength and low dust content, but with a smaller grain size. Its spherical shape makes it preferred for fluidized bed applications such as water filtration.

[0145] Impregnated Carbon

[0146] Porous carbons containing several types of inorganic impregnates such as iodine and silver are impregnated carbon. Cations such as aluminium, manganese, zinc, iron, lithium, and calcium have also been prepared for specific applications. Due to its antimicrobial and antiseptic properties, silver loaded activated carbon is used as an adsorbent for purification of domestic water. Drinking water can be obtained from natural water by treating the natural water with a mixture of activated carbon and aluminium hydroxide (A1(OH)3), a flocculating agent. Impregnated carbons are also used for the adsorption of hydrogen sulphide (H2S) and thiols.

[0147] Polymer Coated Carbon

[0148] This is a process by which a porous carbon can be coated with a biocompatible polymer to give a smooth and permeable coat without blocking the pores. The resulting carbon is useful for various applications including hemoperfusion.

[0149] Woven Carbon

[0150] There is a technology of processing technical rayon fiber into activated carbon cloth for carbon filtering. Adsorption capacity of activated cloth is greater than that of activated charcoal (BET theory) surface area: 500-1500 m2 / g, pore volume: 0.3-0.8 cm3 / g). Owing to the different forms of activated material, it can be used in a wide range of applications. Carbon Nanotubes (CNTs)

[0151] CNTs are cylindrical structures made of carbon atoms arranged in a hexagonal lattice. They have an extremely high surface area and exceptional mechanical, thermal, and electrical properties. CNTs find applications in fields like electronics, materials science, and nanotechnology.

[0152] Graphene

[0153] Graphene is a single layer of carbon atoms arranged in a hexagonal lattice. It has an extremely high surface area and remarkable electronic and thermal conductivity. Graphene is used in various industries, including electronics, energy storage, and composite materials.

[0154] Carbon Aerogels

[0155] Carbon aerogels are lightweight, highly porous materials with a three-dimensional network structure. They are used in applications such as energy storage devices (supercapacitors), catalysis, and as lightweight structural materials.

[0156] Carbon Black

[0157] Carbon black is a fine powder produced by the incomplete combustion of hydrocarbons. It is widely used as a reinforcing filler in rubber products, such as tires, and as a pigment in inks and coatings.

[0158] Carbon Fibers

[0159] Carbon fibers are composed of carbon atoms bonded together in a crystalline structure. They are known for their high tensile strength, low weight, and resistance to heat. Carbon fibers are used in aerospace, automotive, and sporting goods industries to make lightweight and strong composite materials.

[0160] Carbon Nano fibers (CNFs)

[0161] Like carbon nanotubes, carbon nanofibers are cylindrical structures with diameters in the nanometer range. They are used in various applications, including reinforcement in composites, energy storage devices, and catalyst supports. Carbon Cloth and Carbon Paper

[0162] These materials are often made from carbon fibers and are used as electrodes in fuel cells and other energy storage devices. They provide a high surface area for electrochemical reactions.

[0163] Carbon Foam

[0164] Carbon foam is a three-dimensional, open-cell structure made from carbon. It is lightweight and possesses good thermal and electrical conductivity. Carbon foam finds applications in heat exchangers, thermal management systems, and as an electrode material.

[0165] Crosslinked Polymers

[0166] Polymeric Resins with Aromatic Rings

[0167] Polymeric adsorbents containing aromatic rings, such as those based on polystyrene - divinylbenzene (PS / DVB) copolymers, may be used for the adsorption of haloforms. The aromatic nature of these resins enhances their affinity for these compounds.

[0168] Polydimethylsiloxane (PDMS) and Silica-Based Polymers

[0169] PDMS is a silicone-based polymer known with high affinity for haloforms. PDMS- coated fibers or particles, as well as silica-based polymers, may be used in solid-phase microextraction (SPME) for the extraction haloforms.

[0170] Polyacrylate Resins

[0171] Polyacrylate resins are compatible with haloform compounds. They can be tailored to have specific functional groups that enhance their affinity for haloforms. Functional groups of interest include groups containing halides, groups with amide-containing ligands, groups with polar moieties such as hydroxyl and carbonyl groups, and ion exchange groups such as quaternary ammonium and sulfonic groups. Each of these functional groups can enhance the formation of favorable interactions with haloforms.

[0172] Graphene Oxide and Modified Graphene-Based Materials

[0173] Graphene oxide and modified graphene materials may be used for haloform adsorption due to their high surface area and unique properties. Functionalization of graphene can be done to improve selectivity for haloforms. Graphene oxide (GO) and reduced graphene oxide (rGO) can be functionalized with oxygen-containing groups (e.g., hydroxyl, carboxyl) on the basal plane and edges. These groups can enhance interactions with haloforms through hydrogen bonding.

[0174] Introduction of halide-containing functional groups (e.g., chloromethyl or chlorophenyl groups) onto the graphene surface can enhance haloform binding.

[0175] Nitrogen-doping of graphene introduces basic nitrogen functionalities, which interact favorably with haloforms. Nitrogen-containing groups can enhance the overall adsorption capacity of the graphene.

[0176] Optimization of the graphene structure can maximize 71-71 stacking interactions with haloforms. This involves controlling the number of graphene layers or introducing aromatic moieties.

[0177] Incorporation of functionalized carbon nanotubes into graphene -based materials can increase their affinity for haloforms. The tubular structure and functional groups on carbon nanotubes can contribute to improved haloform adsorption.

[0178] Poly(N-vinyl-2-pyrrolidone) (PVP)

[0179] PVP and PVP -based materials are known for their compatibility with a variety of organic compounds. These materials can be used in various forms, such as fdms or particles, for adsorption applications with haloforms.

[0180] Styrene-Divinylbenzene Polymers

[0181] Styrene-divinylbenzene (Styrene-DVB) copolymers with a macroporous structure can also be used for the adsorption of haloforms. These macroporous structures are characterized by pores with diameters typically in the range of 50 nm to several micrometers.

[0182] Amberchrom resins, such as Amberchrom CG161C, are macroporous Styrene-DVB copolymers that are commonly used for chromatographic separations. These resins have a high surface area and porosity.

[0183] Chromabond resins, including Chromabond HR-X, are macroporous copolymers of styrene and divinylbenzene. They are utilized in solid-phase extraction (SPE) for sample preparation and purification.

[0184] XAD resins, such as XAD-4 and XAD-16, are macroporous Styrene-DVB copolymers widely used for adsorption and extraction of organic compounds from air and water samples. Diaion HP (Hypercrosslinked Porous) resins are macroporous Styrene-DVB copolymers manufactured by Mitsubishi Chemical. They are designed for applications such as chromatography and separation processes.

[0185] NeviPure offers macroporous crosslinked polystyrene-divinylbenzene resins for various applications, including the removal of impurities from pharmaceutical intermediates.

[0186] MN200 is a macroporous Styrene-DVB copolymer resin used for adsorption applications, including solid-phase extraction (SPE) and chromatography.

[0187] In certain aspects, the crosslinked polymer comprises a styrene-divinylbenzene group with a macropore structure. In one aspect, the crosslinked polymer is selected from the group consisting of PUROSORB® PAD400, PAD500, PAD600, PAD900, PAD1200, PAD350, PAD610, PAD910, PAD950, PAD950C, Amberlite® FPX66, FPX68, Amberlite® XAD2, XAD4, XAD16, XAD1 180, XAD200, XAD2010, XAD16N, XAD1600N, XAD18, XAD1 180N, XAD7HP, DIADION® Sepabe, XAD761 ® HP20, HP20SS, HP21 , SP70, SP700, SP825L, SP850, CHP20, CHP50, SP207, HP2MGL, LEWATIT® AF 5, SEPLITE® CT10, LX20, LX 207, LXA8, LXA10, LXA17, LXA680, LXA1600, LXA1 180, LXA81, LXA816, LXA817, LXA8302, LXA88, LXS868, and AB-8. In another aspect, the crosslinked polymer is PUROSORB® PAD600 (polydivinylbenzene macroporous, adsorbent resin, non-ionic form).

[0188] In other aspects, the crosslinked polymer is a carbonaceous material, polystyrene, polyacrylic, polyacrylic ester, cation exchange resin, or polystyrene-divinylbenzene. Nonlimiting examples include Amberlite (Rohm and Haas) XAD-2, XAD-4, XAD-7, XAD-16, XAD-18, XAD-1180, XAD-1600, XAD-2000, XAD-2010; Amberchrom (Toso Haas) CG- 71m, CG-71c, CG-161m, CG161c; Diaion Sepabeads (Mitsubishi Chemicals) HP20, SP206, SP207, SP850, HP2MG, HP20SS, SP20MS; Dowex (Dow Chemical) XUS-40285, XUS- 40323, XUS-43493 (also referred to as Optipore V493 (dry form) or Optipore L493 (hydrated form)), Optipore V503, Optipore SD-2; Hypersol Macronet (Purolite) MN-100, MN-102, MN- 150, MN- 152, MN- 170, MN-200, MN-202, MN-250, MN-252, MN-270, MN-300, MN-400, MN-500, MN-502, Purosorb (Purolite) PAD 350, PAD 400, PAD 428, PAD 500, PAD 550, PAD 600, PAD 700, PAD 900, and PAD 950.

[0189] Modification of High Surface Area Materials

[0190] High surface area materials containing an aromatic ring, such as cross-linked polystyrene or graphene, may be modified to add one or more functional groups to one or more of their aromatic rings. Functional groups such as formyl groups may be added to one or more benzene rings of the high surface area materials via Rieche formylation. The one or more formyl groups may then be converted into trihaloketone groups, such as tribromomethylketone groups. For example, it has been shown that cross-linked divinylbenzene or polystyrene / divinylbenzene can be functionalized through a formylation reaction (Macromolecules, 1991, 24, 20-22) and subsequently converting the formyl groups to tribromomethylketone groups (Tetrahedron Letters, Vol. 33, No. 24, 3435-3438). By functionalizing the high surface area material, the high surface area material itself can serve as a prodrug carrier for bromoform where it will hydrolyze in the rumen to release bromoform. An example of a synthetic route to such a material is given below.

[0191] Animal Feed

[0192] Also disclosed herein is combination of an animal feed and the anti-methanogenic composition(s) described herein. The animal feed itself may be solid (e.g., powder, granules, pellets, lick block), semi-solid (e.g. gel, ointment, cream, paste), liquid (e.g. solutions, suspensions, emulsions), or any combination thereof. The anti-methanogenic composition being added to the animal feed may independently be solid (e.g., powder, granules, pellets), semi-solid (e.g., gel, ointment, cream, paste) or liquid (e.g. solutions, suspensions, emulsions). For example, the animal feed and the anti-methanogenic composition to be added to the feed may both be liquid or both be semi-solid or both be solid. Alternatively, the animal feed and composition may each be in a different physical state. For example, the animal feed may be solid or semi-solid and the anti-methanogenic composition may be liquid. The anti- methanogenic composition may, for example, be used to "top-dress" (added on top) a ruminant feedlot ration or may be used to blend into a total mixed ration. In one aspect, the animal feed is in the form of a feed block. In another aspect, the animal feed is in the form of a lick block. In another aspect, the animal feed is in the form of a low moisture block.

[0193] The anti-methanogenic composition may, for example, be added to the drinking water of the animal. In certain embodiments, the anti-methanogenic composition may 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.

[0194] The three main types of animal feed include roughages, concentrates and mixed feeds. In general, roughages contain a higher percentage of crude fiber and a lower percentage of digestible nutrients than concentrates. For example, roughages may be defined as containing equal to or greater than 20 wt.% crude fiber and equal to or less than 60 wt.% total digestible nutrients. Roughages may include, for example, dry roughages (e.g., hay, straw, artificially dehydrated forages containing at least 90 wt.% dry matter), silages (formed from green forages such as grass, alfalfa, sorghum, and corn and preserved in a silo at dry matter contents of 20 to 50 %), and pastures (e.g. green growing pastures providing forage that has a high water content and generally less than 30 % dry matter). The two basic types of roughages include grasses and legumes. Grasses are generally higher in fiber and dry matter than legumes. Legumes are generally higher in proteins, metabolizable energy, vitamins and minerals. Concentrates contain a relatively lower percentage of crude fiber and a higher percentage of digestible nutrients than roughages. For example, concentrates may be defined as containing less than 20 wt.% crude fiber and greater than 60 wt.% total digestible nutrients. Concentrates may include, for example, energy-rich grains and molasses. Corn, wheat, oats, barley and milo (sorghum grain) are energy-rich grains, containing about 70 to 80 wt.% total digestible nutrients.

[0195] Mixed feeds are generally a mixture of roughages and concentrates to provide "complete" balanced rations and may be either high or low in energy, protein or fiber. The disclosed compositions, for example, can be combined with animal feed in various amounts depending on the total amount of the composition intended to be administered to the animal.

[0196] The animal feed may, for example, comprise from about 0.0001 wt.% to about 10 wt.% of the disclosed compositions depending on the loading level of the active ingredient (e.g., bromoform), based on the total dry weight of the animal feed. The animal feed may, for example, comprise from about 0.01 wt.% to about 10 wt.% of disclosed composition, based on the total dry weight of the animal feed. For example, the animal feed may comprise from about 0.001 wt.% to about 9.5 wt.%, or from about 0.005 wt.% to about 9 wt.%, or from about 0.01 wt.% to about 8.5 wt.%, or from about 0.05 wt.% to about 8 wt.%, or from about 0.1 wt.% to about 7.5 wt.%, or from about 0.9 wt.% to about 7 wt.%, or from about 1 wt.% to about 6 wt.%, or from about 1.5 wt.% to about 5.5 wt.%, or from about 2 wt.% to about 5 wt.%, or from about 2.5 wt.% to about 4.5 wt.%, or from about 3 wt.% to about 4 wt.% disclosed composition based on the total dry weight of the animal feed. For example, the animal feed may comprise from about 0.4 wt.% to about 9.5 wt.%, or from about 0.5 wt.% to about 9 wt.%, or from about 0.6 wt.% to about 8.5 wt.%, or from about 0.7 wt.% to about 8 wt.%, or from about 0.8 wt.% to about 7.5 wt.%, or from about 0.9 wt.% to about 7 wt.%, or from about 1 wt.% to about 6 wt.%, or from about 1.5 wt.% to about 5.5 wt.%, or from about 2 wt.% to about 5 wt.%, or from about 2.5 wt.% to about 4.5 wt.%, or from about 3 wt.% to about 4 wt.% disclosed composition based on the total dry weight of the animal feed.

[0197] In one embodiment, the disclosed composition is administered at a dose of preferably at least 16.67, 10, 5, 3, 2, 1, 0.5, 0.25 0.125 or 0.067% of the organic matter administered to the ruminant animal. For example, if a 450 kg ruminant animal (e.g., steer) consumes 2.5% to 3% of its body weight per day of feed, then the disclosed composition is administered at a dose proportional to the amount of organic matter administered to the ruminant. In the case of a 450 kg ruminant animal, and where 80% of the feed is organic matter, if the animal consumes about 2.5% of its body weight per day, then the disclosed composition is administered at a dose of about 0.27, 0.18, 0.09, 0.045, 0.0225, 0.01125 or 0.00603 kg per day to result in a dose at least 3, 2, 1, 0.5, 0.25 0.125 or 0.067% of the organic matter administered to the ruminant animal.

[0198] Some anti-methanogenic compositions disclosed herein are used as an animal feed additive. Certain anti-methanogenic compositions disclosed herein are used as a nutritional preparation for an animal.

[0199] Use of the anti-methanogenic compositions in an effective amount in the animal feed additives or the nutritional preparations disclosed herein reduce methane emission of a ruminant.

[0200] Advantages Provided by the Slow Release of the Anti-Methanogenic Composition

[0201] In certain aspects, the anti-methanogenic composition is a slow-release formulation for inhibiting methane production in ruminants, particularly cattle, and offers several advantages in addressing both environmental and practical concerns. For the prodrug, the slow-release characteristics are determined by the rate of the hydrolysis reaction which leads to the release of the haloform. This rate of release may in turn be affected by the rumen pH. The anti- methanogenic composition delivers bromoform which inhibits methane production by disrupting the enzymatic pathways used by methanogenic archaea in the rumen. The slow- release formulation amplifies its effectiveness for at least the following reasons: Sustained Methane Inhibition

[0202] The slow-release formulation allows for a continuous and gradual release of bromoform into the rumen. This maintains a consistent level of the compound over time, which more effectively suppresses methane production compared to a single large dose, where the effect might wear off too quickly.

[0203] Reduced Toxicity and Safety Concerns

[0204] Bromoform can be toxic at high concentrations, so a slow-release formulation minimizes the risk of delivering too much bromoform at once. By releasing small amounts over time, the risk of toxicity to the animal is reduced, making it a safer option for long-term methane mitigation in livestock farming.

[0205] Improved Digestive Health and Function

[0206] The slow-release formulation avoids any sudden disruption of the microbial balance in the rumen, which could otherwise affect the digestion of fibrous feed. Maintaining rumen health is crucial for the efficient digestion of feed and for the overall well-being of the animal.

[0207] Extended Efficacy with Less Frequent Dosing

[0208] The slow-release formulation reduces the need for frequent supplementation, making it more practical for farmers to use. This is especially important in large-scale livestock farming, where daily administration of additives can be labor-intensive and costly. The slow-release product lasts for weeks or months, thereby reducing the need for constant intervention.

[0209] Reduced Methane Emissions Over the Long Term

[0210] Consistent inhibition of methane production over time, thanks to the slow-release formulation, has a significant cumulative impact on reducing greenhouse gas emissions from the agricultural sector. This approach helps meet sustainability goals by ensuring that methane suppression is effective over a longer period.

[0211] Minimal Impact on Feed Intake and Animal Productivity

[0212] When bromoform is administered in a form that releases gradually, it is less likely to alter the palatability of the feed or the animal's appetite. Maintaining proper feed intake and avoiding disruption to animal productivity (such as weight gain and milk production) are critical for farmers, and the slow-release formulation helps in this regard. Cost-Effective and Efficient

[0213] The slow-release formulation provides a relatively low-cost solution with less frequent dosing and more effective long-term methane suppression. This makes it an attractive option for large-scale adoption in the livestock industry.

[0214] Any embodiment of the invention is meant to be illustrative only and is not meant to be limiting to the invention. Therefore, it should be appreciated that various other changes and modifications can be made to any embodiment described without departing from the spirit and scope of the invention.

[0215] All headings are for the convenience of the reader and should not be used to limit the meaning of the text that follows the heading, unless so specified.

[0216] The present invention is further illustrated by the following examples that should not be construed as limiting. The contents of all references, patents, and published patent applications cited throughout this application, as well as the Figures, are incorporated herein by reference in their entirety for all purposes.

[0217] EXAMPLES

[0218] Example 1. Hydrolysis of Tribromopyruvyl Urea Produces Bromoform under Conditions Similar to Those in the Rumen

[0219] Background

[0220] Trihalomethyl ketones are relatively stable compounds that can be hydrolyzed to produce bromoform under conditions common in the rumen (see FIG. 1). A trihalomethyl ketone of particular interest is tribromopyruvyl urea (also known as 3,3,3-tribromo-A- carbamoyl-2-oxopropanamide), which can be synthesized using the general scheme outlined in FIG. 2. The Inventors used tribromopyruvyl urea as a non-limiting example of the stable trihalomethyl ketones that can be administered to ruminant animals and accurately dosed to inhibit methane production.

[0221] Alcohols and amines also react with trihalomethyl ketones as shown in FIG. 3 to generate trihalomethanes. Two non-limiting examples of synthetic schemes for preparing tribromomethyl ketones are presented in FIGs. 4 and 5. The synthetic scheme presented in FIG.4 can be used to prepare other trihalomethyl ketones (e.g., triiodomethyl ketones and trichloromethyl ketones) by replacing the tribromomethyl alcohol with a triiodomethyl alcohol or a trichloromethyl alcohol. Materials and Methods

[0222] Synthesis of Bromoform Precursor

[0223] 150 grams of maleic anhydride (1.53 moles) and 90 g of urea (1.50) moles were mixed in 300 mL of glacial acetic acid at 50 °C for 12 hours. During this process, the compounds first dissolved and then a precipitate started to form. After 12 hours, the reaction was cooled to 0 °C for 48 hours resulting in the formation of more precipitate. This precipitate was filtered, washed with cold acetic acid and then dried to yield approximately 170 grams of maleuric acid (70 % yield).

[0224] 1.58 grams of maleuric acid (10 mmol) was first added to 100 mL of water. With vigorous magnetic stirring, approximately 2.8 grams of bromine (35 mmol) was added, and the solution was heated at 30 to 35 °C for 6 hours. Following this, sodium thiosulfate was added to eliminate excess bromine, and the solution was cooled to 0 °C for 12 hours. The precipitates were fdtered, washed with cold water, and then dried at 50 °C resulting in 1.18 grams of tribromopyruvyl urea (32 % yield).

[0225] Bromoform Release Testing

[0226] A 100 mM solution of monopotassium dihydrogen phosphate was made, and the pH adjusted using potassium hydroxide to yield five solutions with pH values of 4.34, 6.71, 7.71, 11.2 and 12.32. To 10 mL of each of these solutions, approximately 30 mg of dried tribromopyruvyl urea was added into a 20 mL glass vial with a sealed septum cap. This suspension was placed in an oven at 40 °C and occasionally the vials were shaken. After a period of time, 100 LIL of each of the solutions was extracted and diluted into a septum capped headspace vial containing 10 mL of a pH 4.3 potassium phosphate buffer solution. The bromoform content of these solutions was analyzed using GCMS to measure the conversion of tribromopyruvyl urea to bromoform.

[0227] GCMS Analysis

[0228] Gas chromatography-mass spectroscopy (GCMS) was used to analyse the bromoform content in water solutions and in the stabilized solid samples that are the subject of this disclosure. A headspace technique was used where the sample was placed inside of a 20 mL sample vial with a silicone septum and exposed to an elevated temperature. When the sample reached equilibrium after a period, the vial was pressurized with helium gas and then a portion of this volume was analyzed by GCMS. To quantify the amount of material, the temperature and volume of liquid were chosen such that the concentration in the headspace at elevated temperature was proportional to the amount in the original unheated sample. For bromoform in water, 10 mL of solution was used, and the temperature used was 40 °C. At these elevated temperatures, the bromoform in its respective matrices demonstrated a linear response using a standard addition method and the generation of numerous calibration curves.

[0229] A Shimadzu GC-2030 gas chromatograph was used in conjunction with a HS-20NX headspace sampler module and a GCMS-QP2020NX single quadrupole mass spectrometer. A Shimadzu SH-I-5SH column with a silylene phase comprising a 5% diphenyl and 95% dimethyl polysiloxane was used. This column was chosen for its ability to separate various halomethanes (e.g., chloroform, bromochloromethane). For the headspace analyzer, the oven temperature used was dependent on the matrix (see above). Sample line and transfer line temperatures of 150 °C and 150 °C, respectively, were used. A shaking level of 4 was used for a 20-minute equilibrating time. The sample was pressurized for 1 minute to 55.2 kPA with helium and allowed to equilibrate for 0.50 minutes. Load and load equilibration times of 0.50 and 0.10 minutes, respectively, were used. An injection time of 1.50 minutes was used followed by a 4-minute needle flush time.

[0230] The GC oven temperature was set to 50 °C initially and ramped to 150 °C at a 35 °C / minute rate, then to 200 °C at a 50 °C / minute rate, and finally to 250 °C at a 70 °C / minute rate. A 50-ratio split injection was used with a gas pressure of 147.0 kPa and total column flow of 54.0 mL / minute (comprising 1.00 mL / minute column flow and 3.0 mL / minute purge flow). For the mass spectrometer, ion source and interface temperatures of 230 °C and 220 °C, respectively, were used. The spectrometer was tuned using perfluorotributylamine to maximize the m / z 502 peak. A detector voltage of -0.1 kV relative to this tuning was used for this analysis. A linear calibration curve was established using 10 mL of aqueous solution at concentrations of 0, 12.5, 25, 37.5, 50, 62.5, 75, 87.5 and 100 pM.

[0231] Results and Discussion

[0232] We observed significant hydrolysis yielding bromoform for pH values of 4.3 to 7.7 which covers the range of likely rumen pH values (typically 6.2 to 7) in healthy ruminants (see Table 1). Table 1. Production of bromoform from hydrolysis of tribromopyruvyl urea in aqueous solution at increasing pH conditions.

[0233] The literature on hydrolysis of tribromomethyl ketones suggests that a more alkaline environment is conducive to faster hydrolysis yielding bromoform. Unexpectedly, at longer hydrolysis times under alkaline conditions, the concentration decreases from 24 hours to 48 hours. This suggests that there might be a slow secondary reaction with urea that is taking place. Regardless, these alkaline conditions are not indicative of rumen pH values.

[0234] Example 3. Assessment of Stabilized Formulations with In Vitro Fermentation Technique

[0235] The in vitro fermentation technique (IVFT) was conducted as described in a previous publication (Journal of the Science of Food and Agriculture, 2014; 94: 1191-1196). Briefly, Bellco tubes were set up with 0.1 g of high fiber fermentation substrate (oaten chaff, ground to pass 1 mm screen) and placed inside an anaerobic chamber (80% N2 / 10 % CO2 / 10% H2). The required amount of either pure bromoform stock or raw material stocks was added to the Bellco tube containing fermentation substrate. Following this, buffered sheep rumen fluid (10 mL) was added, and the tubes were sealed and crimped. A substrate control (oaten chaff only), and an inoculum control (rumen fluid only) were included, and all treatments were run in triplicate. Tubes were then taken out of the chamber and incubated at 39°C for 24 hours with orbital shaking at 50 rpm. At the end of incubation, tubes were taken out, and total microbial gas produced was measured using a pressure transducer, as a proxy of overall microbial activity, before a sample of headspace gas was collected for methane analysis by gas chromatography. For this study, DMSO solutions of tribromopyruvyl urea were used to aid in dosing. DMSO was chosen as it is relatively benign biologically at these levels, it readily dissolves tribromopyruvyl urea, it is fully miscible in water, and it will not hydrolyze the tribromopyruvyl urea prematurely. As seen in Table 2, at nominal tribromopyruvyl urea concentrations of 2.5, 5, 7.5 and 10 pM, almost complete inhibition of methane production was observed. At a concentration of 1.0 pM, the inhibition relative to the control is about 20%. This is further confirmation that the tribromopyruvyl urea will slowly release bromoform into the rumen where it can inhibit methane production.

[0236] Table 2. Methane and gas production (mean ± SE) when the treatments were combined with oaten chaff substrate in the IVFT. Each treatment was tested in six replicates. Within the same column, treatments not connected by the same letter are significantly different (p<0.05). Percentage reduction when compared to carrier control, negative percentage reduction values indicate an increase. DMi = dry matter incubated.

[0237] CH4

[0238] Total gas concentration CH4yield

[0239] Treatment % reduction volume (mL) (mL / 100 mL (mL / g DMi) gas)

[0240] Total CH4CH4gas cone. yield

[0241] Carrier 134.7a± 0.75 10.5a± 0.08 28.3a± 0.37

[0242] 1 LIM 133. lab± 1.02 8.9b± 0.15 23.7b± 0.35 3 15 16

[0243] 2.5 pM 130.1b± 0.45 0.5c± 0.01 1.2c± 0.03 7 96 96

[0244] 5 LIM 133.4b± 0.76 0.4c± 0.01 1.2c± 0.02 3 96 96

[0245] 7.5 pM 135.8a± 0.71 0.4c± 0.01 l.lc± 0.01 -2 96 96

[0246] 10 pM 134.8a± 0.74 0.4c± 0.01 l.lc± 0.03 0 96 96

[0247] While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains and as may be applied to the essential features hereinbefore set forth.

[0248] INCORPORATION BY REFERENCE

[0249] All references, articles, publications, patents, patent publications, and patent applications cited herein within the above text and / or cited below are incorporated by reference in their entireties for all purposes. However, mention of any reference, article, publication, patent, patent publication, and patent application cited herein is not, and should not be taken as acknowledgment or any form of suggestion that they constitute valid prior art or form part of the common general knowledge in any country in the world.

[0250] REFERENCES

[0251] Balch, W. E. & Wolfe, R. S. 1976. Appl Environ Microbiol, 32, 781-91.

[0252] Cavallito, Chester J., and Clayton S. Smith, Journal of the American Chemical Society 63.4 (1941): 995-998.

[0253] Dohi, Souya, Katsuhiko Moriyama, and Hideo Togo. European Journal of Organic Chemistry 2013.34 (2013): 7815-7822.

[0254] Gagen, E. J., Wang, J., Padmanabha, J., Liu, J., De Carvalho, I., Liu, J., Webb, R. L, Al Jassim, R., Morrison, M., Denman, S. E. & Mcsweeney, C. S. 2014. BMC Microbiol, 14, 314.

[0255] Gulizhabaier, A., and A. A. Rexit, Russian Journal of Organic Chemistry 57.5 (2021): 809-815.

[0256] Hasimujiang, Balati, et al. Chemistry Select 4.19 (2019): 5560-5562.

[0257] Hasimujiang, Balati, et al. Synthetic Communications 48.8 (2018): 887-891.

[0258] Honan, M., Feng, X., Tricarico, J.M. 2020. Animal Production Science 62(14) 1303- 1317.

[0259] Huang R. et al., Animal, 2023, 17(5), pp.100788.

[0260] Jayaraman, Aravindan, et al., Advanced Synthesis & Catalysis 360.20 (2018): 3978- 3989.

[0261] Maczulak, A. E., Wolin, M. J. & Miller, T. L. 1989. Appl Environ Microbiol, 55, 2468- 73.

[0262] Miller, T. L., Wolin, M. J., De Macario, E. C. & Macario, A. J. 1982. Applied and Environmental Microbiology, 43, 227-232.

[0263] Morgavi DP et al., Animal. 2023 Jul; 17 Suppl 3:100830.

[0264] Patra AK, Environ Monit Assess. 2012 Apr;184(4):1929-52.

Claims

CLAIMSWhat is claimed is:

1. An anti-methanogenic composition comprising: a trihalomethyl ketone having a structure of Formula (I):whereinR1represents hydrogen, substituted or unsubstituted Ci-Ce alkyl such methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl, neopentyl, isopentyl, pentan-2-yl, 3-pentyl, sec-isopentyl, 2-methylbutyl, hexyl, isohexyl, neohexyl, terthexyl, hexan-2-yl, hexan-3-yl, 2-methylpentyl, 3 -methylpentyl, 2,3-dimethylbutyl, substituted or unsubstituted Cs-C'x cycloalkyl or Ci-Cs-alkyl-Cs-Cs-cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopropyl-methyl, cyclobutylmethyl, cyclopentyl-methyl, cyclohexyl-methyl, cycloheptyl-methyl, cyclooctyl-methyl, cyclopropyl-ethyl, cyclobutyl-ethyl, cyclopentyl-ethyl, cyclohexyl-ethyl, cycloheptyl-ethyl, cyclooctyl-ethyl, cyclopropyl-propyl, cyclobutyl-propyl, cyclopentyl-propyl, cyclohexylpropyl, cycloheptyl-propyl, cyclooctyl-propyl, substituted or unsubstituted C5-C14 aryl or Ci- C3-alkyl-C5-Ci4-aryl, -(CO)NR3(CO)NR4R5;R2represents halogen; andR3, R4, and R5each independently represents hydrogen, halogen, substituted or substituted C1-C4 alkyl such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tertbutyl, or combinations thereof; and optionally an agriculturally acceptable carrier.

2. The anti-methanogenic composition of claim 1, wherein R2represents Cl or Br.

3. The anti-methanogenic composition of claim 1 or 2, wherein R2represents Br.

4. The anti-methanogenic composition of any one of claims 1 to 3, wherein R1is substituted by one or more halogens, -OH, -COOH, -NO2, -OCH3, -CH3, -CN, or -NH2.

5. The anti-methanogenic composition of any one of claims 1 to 3, wherein R1represents -(CO)NR3(CO)NR4R5.

6. The anti-methanogenic composition of claim 5, wherein the trihalomethyl ketone is Formula (1-1):

7. The anti-methanogenic composition of any one of claims 1 to 6, wherein the agriculturally acceptable carrier is a hydrophobic carrier.

8. The anti-methanogenic composition of claim 7, wherein the agriculturally acceptable carrier comprises one or more of a vegetable oil, an animal fat, and / or a microbial oil, and / or a mixture thereof.

9. The anti-methanogenic composition of claim 8, wherein the vegetable oil comprises one or more of coconut oil, corn oil, cottonseed oil, olive oil, palm oil, peanut oil, canola oil, safflower oil, sesame oil, soybean oil, sunflower oil, almond oil, beech nut oil, Brazil nut oil, cashew oil, Jamaican cobnut oil, hazelnut oil, macadamia oil, mongongo nut oil, pecan oil, pine nut oil, pistachio oil, walnut oil, pumpkin seed oil, grapefruit seed oil, lemon oil, orange oil, avocado oil, poppyseed oil, tea seed oil, and / or apricot oil, and / or a mixture thereof.

10. The anti-methanogenic composition of claim 8, wherein the animal fat comprises one or more of fish oil, pig fat, chicken fat, cow fat, and / or butter, and / or a mixture thereof.

11. The anti-methanogenic composition of claim 8, wherein the microbial oil comprises oils produced by one or more of Yarrowia lipoytica, Schizochytrium sp., Crypthecodinium cohnii, Mortierella alpina, Mortierella elongate, Mortierella isabelline, Porphyridium cruentum, Mucor circinelloides , Aspergillus niger, Borago officinalis, Nitzschi laevis, Navicula pelliculosa, Cylindrotheca fusiformis, Scendesmus obtusiusculus , and / or Haema-tococcus pluvialis, and / or a mixture thereof.

12. The anti-methanogenic composition of any one of claims 1 to 11, wherein the trihalomethyl ketone is amphiphilic.

13. The anti-methanogenic composition of any one of claims 1 to 12, wherein the trihalomethyl ketone has a solubility to water (mWater / mFomiuia(i)) of from about 0.1 to about 100, from about 0.5 to about 100, from about 1 to about 100, from about 1.25 to about 50, from about 1.5 to about 40, from about 1.75 to about 30, from about 2 to about 20, from about 2 to about 10, or from about 5 to about 10.

14. The anti-methanogenic composition of any one of claims 1 to 13, wherein the trihalomethyl ketone has a solubility to food oil (mWater / moii) of from about 0.1 to about 100, from about 0.5 to about 100, from about 1 to about 100, from about 1.25 to about 50, from about 1.5 to about 40, from about 1.75 to about 30, from about 2 to about 20, from about 2 to about 10, or from about 5 to about 10.

15. The anti-methanogenic composition of any one of claims 1 to 14, wherein the trihalomethyl ketone has a viscosity under standard atmospheric conditions (25°C and pressure of 1 bar) of about 0.250 mPa-s to about 1000 mPa-s, about 0.500 mPa-s to about 1000 mPa-s, about 0.750 mPa-s to about 1000 mPa-s, about 1 mPa-s to about 1000 mPa-s, about 2.5 mPa-s to about 1000 mPa-s, about 5 mPa-s to about 1000 mPa-s, about 7.5 mPa-s to about 1000 mPa-s, about 10 mPa-s to about 1000 mPa-s, about 15 mPa-s to about 1000 mPa-s, about 20 mPa-s to about 1000 mPa-s, about 25 mPa-s to about 1000 mPa-s, about 25 mPa-s to about 750 mPa-s, about 25 mPa-s to about 500 mPa-s, about 25 mPa-s to about 250 mPa-s, about 25 mPa-s to about 100 mPa-s, about 25 mPa-s to about 75 mPa-s, or about 25 mPa-s to about 50 mPa-s.

16. The anti-methanogenic composition of any one of claims 1 to 15, further comprising a bulking agent, wherein the bulking agent comprises polysaccharides.

17. The anti-methanogenic composition of claim 16, wherein the polysaccharides comprise cellulose, hemicellulose, starch, inulin, chitin, and / or pectin, and / or a mixture thereof.

18. The anti-methanogenic composition of claim 17, wherein: cellulose is derived from one or more of grass (such as Bermuda grass, brome, oatgrass, fescue, false oat grass, meadow grasses, orchard grass, ryegrass, and timothy-grass), millet, oats, sorghum, soybean, cotton, hemp, flax, jute, corn (such as corn cob, maize straw), banana, wheat (such as wheat straw), cork bark, curaua, kenaf, nettle, ramie, rice husk, hardwood, softwood, sugar can bagasse, sisal, regenerated celluloses (such as viscose and cellophane), and / or bacterial cellulose (produced by species such as Acetobacter pasteurianum, Acetobacter xylinum, Acetobacter rancens, Komagataeibacter xylinus, Sarcina ventriculi, and / or Bacterium xylinoides , hemicellulose is derived from one or more of grass (such as Bermuda grass, brome, oatgrass, fescue, false oat grass, meadow grasses, orchard grass, ryegrass, and timothy-grass), millet, oats, sorghum, soybean, cotton, hemp, flax, jute, corn (such as corn cob, maize straw), banana, wheat (such as wheat straw), cork bark, curaua, kenaf, nettle, ramie, rice husk, hardwood, softwood, sugar can bagasse, and / or sisal; starch is derived from one or more of rice, wheat, maize, potato, cassava, acorn, arrowroot, arracacha, banana, barley, breadfruit, buckwheat, canna, colocasia, cuckoo-pint, katakuri, kudzu, malanga, millet, oat, oca, Polynesian arrowroot, sago, sorghum, sweet potatoe, rye, taro, chestnut, water chestnut, yam, sorghum, soybean, fava, lentil, mung bean, adzuki bean, pea, and / or chickpea; inulin is derived from one or more of Agave, banana, burdock, camas, chicory, coneflower, costus, dandelion, elecampane, garlic, globe artichoke, Jerusalem artichoke, jicama, leopard's bane, mugwort, onion, yam, and / or daisy; chitin is derived from fungi and / or arthropods; pectin is derived from one or more of pear, apple, guava, quince, plum, citrus (such as orange, lemon, grapefruit, pomelo, lime), apricot, cherry, carrot, and / or rose hip.

19. The anti-methanogenic composition of any one of claims 16 to 18, wherein the polysaccharides are functionalized with a trihalomethyl ketone through an ether linkage, anamine linkage, a thioether linkage, an ester linkage, a thioester linkage, an amide linkage, a thioamide linkage, or a thionoester linkage.

20. The anti-methanogenic composition of any one of claims 1 to 19, wherein the bulking agent comprises one or more high surface area materials.

21. The anti-methanogenic composition of claim 20, wherein the one or more high surface area materials have a surface area of 50-5000 m2 / g, optionally 100-5000 m2 / g, optionally 200-5000 m2 / g, optionally 300-5000 m2 / g, optionally 500-5000 m2 / g, optionally 500-4000 m2 / g, optionally 500-3000 m2 / g, optionally 750-3000 m2 / g, optionally 900-3000 m2 / g, optionally 1000-3000 m2 / g, optionally 500-1500 m2 / g.

22. The anti-methanogenic composition of any one of claims 20 to 21, wherein the one or more high surface area materials comprise activated carbons and / or crosslinked polymers, and / or a mixture thereof, wherein the activated carbons comprise one or more of: powdered activated carbon (PAC), granular activated carbon (GAC), extruded activated carbon (EAC), bead activated carbon (BAC), impregnated carbon, polymer coated carbon, woven carbon, carbon nanotubes (CNTs), graphene, carbon aerogels, carbon black, carbon fibers, carbon nanofibers (CNFs), carbon cloth, carbon paper, and / or carbon foam, wherein the crosslinked polymers comprise one or more of: polydimethylsiloxane (PDMS) and silica-based polymers, polyacrylate resins, graphene oxide and modified graphene materials, poly(N-vinyl-2-pyrrolidone) (PVP) and PVP -based materials, styrene - divinylbenzene (Styrene-DVB) copolymers, amberchrom resins, chromabond resins, XAD resins, polystyrene, polyacrylic, polyacrylic ester, and / or cation exchange resin.

23. The anti-methanogenic composition of claim 22, wherein one or more aromatic rings of the one or more high surface area materials are modified to add one or more tribromomethylketone functional groups.

24. The anti-methanogenic composition of any one of claims 1 to 6, wherein the trihalomethyl ketone has a partition coefficient (log P), as defined under an octanol-water system, of from about -2.0 to about 2.5, from about -1.75 to about 2.5, from about -1.5 to about 2.5, from about -1.25 to about 2.5, from about -1 to about 2.5, from about -1 to about 2.25,from about -1 to about 2, from about -1 to about 1.75, from about -1 to about 1.5, from about - 1 to about 1.25, from about -1 to about 1, from about -0.75 to about 1, from about -0.5 to about 1, from about -0.5 to about 0.75, or from about -0.5 to about 0.5.

25. Animal feed comprising the anti-methanogenic composition of any one of claims 1 to 24.

26. The animal feed of claim 25, wherein the animal feed is in the form of a powder, granule, pellet, feed block, lick block, gel, ointment, cream, paste, solution, suspension, or emulsion.

27. The animal feed of claim 25 or 26, further comprising a roughage, cereal, starch, vegetable waste, vitamin, mineral, trace element, emulsifier, aromatizing product, binder, colorant, odorant, thickening agent, or a combination thereof.

28. The animal feed of any one of claims 25 to 27, wherein the animal feed comprises from about 0.0001 wt.% to about 10 wt.%, from about 0.001 wt.% to about 9.5 wt.%, from about 0.005 wt.% to about 9 wt.%, from about 0.01 wt.% to about 8.5 wt.%, from about 0.05 wt.% to about 8 wt.%, from about 0.1 wt.% to about 7.5 wt.%, from about 0.9 wt.% to about 7 wt.%, from about 1 wt.% to about 6 wt.%, from about 1.5 wt.% to about 5.5 wt.%, from about 2 wt.% to about 5 wt.%, from about 2.5 wt.% to about 4.5 wt.%, or from about 3 wt.% to about 4 wt.% the anti-methanogenic composition based on the total dry weight of the animal feed.

29. The animal feed of claim 28, wherein the animal feed comprises from about 0.4 wt.% to about 9.5 wt.%, from about 0.5 wt.% to about 9 wt.%, from about 0.6 wt.% to about 8.5 wt.%, from about 0.7 wt.% to about 8 wt.%, from about 0.8 wt.% to about 7.5 wt.%, from about 0.9 wt.% to about 7 wt.%, from about 1 wt.% to about 6 wt.%, from about 1.5 wt.% to about 5.5 wt.%, from about 2 wt.% to about 5 wt.%, from about 2.5 wt.% to about 4.5 wt.%, or from about 3 wt.% to about 4 wt.% the anti-methanogenic composition based on the total dry weight of the animal feed.

30. A method for reducing methane emissions from a ruminant comprising:administering to the ruminant a composition comprising a trihalomethyl ketone having a structure of Formula (I):whereinR1represents hydrogen, substituted or unsubstituted Ci-Ce alkyl such methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl, neopentyl, isopentyl, pentan-2-yl, 3-pentyl, sec-isopentyl, 2-methylbutyl, hexyl, isohexyl, neohexyl, terthexyl, hexan-2-yl, hexan-3-yl, 2-methylpentyl, 3 -methylpentyl, 2,3-dimethylbutyl, substituted or unsubstituted Cs-C'x cycloalkyl or Ci-Cs-alkyl-Cs-Cs-cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopropyl-methyl, cyclobutylmethyl, cyclopentyl-methyl, cyclohexyl-methyl, cycloheptyl-methyl, cyclooctyl-methyl, cyclopropyl-ethyl, cyclobutyl-ethyl, cyclopentyl-ethyl, cyclohexyl-ethyl, cycloheptyl-ethyl, cyclooctyl-ethyl, cyclopropyl-propyl, cyclobutyl-propyl, cyclopentyl-propyl, cyclohexylpropyl, cycloheptyl-propyl, cyclooctyl-propyl, substituted or unsubstituted C5-C14 aryl or Ci- C3-alkyl-C5-Ci4-aryl, or -(CO)NR3(CO)NR4R5;R2represents halogen; andR3, R4, and R5each independently represents hydrogen, halogen, substituted or substituted C1-C4 alkyl such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tertbutyl; and optionally an agriculturally acceptable carrier.

31. The method of claim 30, wherein R2represents Cl or Br.

32. The method of claim 30 or 31 , wherein R2represents Br.

33. The method of any one of claims 30 to 32, wherein R1is substituted by one or more halogens, -OH, -COOH, -NO2, -OCH3, -CH3, -CN, or -NH2.

34. The method of any one of claims 30 to 33, wherein R1represents - (CO)NR3(CO)NR4R5.

35. The method of claim 34, wherein the trihalomethyl ketone is Formula (1-1):

36. The method of any one of claims 30 to 35, wherein the trihalomethyl ketone is administered in an amount of between 1 mg / kg and 1000 mg / kg dry matter intake (DMI) to the ruminant, between 1 mg / kg and 750 mg / kg DMI to the ruminant, between 1 mg / kg and 500 mg / kg DMI to the ruminant, between 1 mg / kg and 400 mg / kg DMI to the ruminant, between 1 mg / kg and 300 mg / kg DMI to the ruminant, between 1 mg / kg and 200 mg / kg DMI to the ruminant, between 1 mg / kg and 100 mg / kg DMI to the ruminant, between 5 mg / kg and 100 mg / kg DMI to the ruminant, between 10 mg / kg and 100 mg / kg DMI to the ruminant, or between 25 mg / kg and 75 mg / kg DMI to the ruminant.

37. The method of any one of claims 30 to 36, wherein the ruminant is a member of the family Bovidae.

38. The method of claim 37, wherein the ruminant is Bos taurus or Bos indicus.

39. The method of claim 37, wherein the ruminant is Ovis aries or Capra hircus.

40. The method of any one of claims 30 to 39, wherein the trihalomethyl ketone is administered orally, parenterally, or by injection.

41. The method of claim 40, wherein the trihalomethyl ketone is administered orally in an animal feed.

42. The method of claim 40 or 41 , wherein the trihalomethyl ketone is administered by a feed dispenser.

43. The method of any one of claims 30 to 42, wherein the methane emissions are reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% compared to an untreated control ruminant.

44. A composition comprising: a trihalomethyl ketone having a structure of Formula (I):whereinR1represents hydrogen, substituted or unsubstituted Ci-Ce alkyl such methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl, neopentyl, isopentyl, pentan-2-yl, 3-pentyl, sec-isopentyl, 2-methylbutyl, hexyl, isohexyl, neohexyl, terthexyl, hexan-2-yl, hexan-3-yl, 2-methylpentyl, 3 -methylpentyl, 2,3-dimethylbutyl, substituted or unsubstituted Cs-C'x cycloalkyl or Ci-Cs-alkyl-Cs-Cs-cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopropyl-methyl, cyclobutylmethyl, cyclopentyl-methyl, cyclohexyl-methyl, cycloheptyl-methyl, cyclooctyl-methyl, cyclopropyl-ethyl, cyclobutyl-ethyl, cyclopentyl-ethyl, cyclohexyl-ethyl, cycloheptyl-ethyl, cyclooctyl-ethyl, cyclopropyl-propyl, cyclobutyl-propyl, cyclopentyl-propyl, cyclohexylpropyl, cycloheptyl-propyl, cyclooctyl-propyl, substituted or unsubstituted C5-C14 aryl or Ci- C3-alkyl-C5-Ci4-aryl, or -(CO)NR3(CO)NR4R5;R2represents halogen; andR3, R4, and R5each independently represents hydrogen, halogen, substituted or substituted C1-C4 alkyl such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tertbutyl; and optionally an agriculturally acceptable carrier.

45. The composition of claim 44, wherein R2represents Cl or Br.

46. The composition of claim 44 or 45, wherein R2represents Br.

47. The composition of any one of claims 44 to 46, wherein R1is substituted by one or more halogens, -OH, -COOH, -NO2, -OCH3, -CH3, -CN, or -NH2.

48. The composition of any one of claims 44 to 47, wherein R1represents - (CO)NR3(CO)NR4R5.

49. The composition of claim 48, wherein the trihalomethyl ketone is Formula (I-1):

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