Methane emission-reducing composition

Abstract

Climate change is an increasing problem creating long-term shifts in temperatures and weather patterns. The disclosure relates to a method of producing a plurality of halogenated carbon compounds, the method comprising contacting a source of electron-rich carbon compounds with a composition comprising hypohalite ions under conditions and for a time sufficient to produce the plurality of halogenated carbon compounds. The disclosure also relates to a composition for producing the halogenated carbon compounds, uses of the halogenated carbon compounds in supplements and feed, and a method of reducing methane emissions of an animal.

Description

METHANE EMISSION-REDUCING COMPOSITIONPRIORITY DOCUMENT

[0001] The present application claims priority from Australian Provisional Patent Application No. 2023901838 titled “Methane Emission-Reducing Composition” and filed on 9 June 2023, the content of which is hereby incorporated by reference in its entirety.TECHNICAL FIELD

[0002] The present disclosure relates to the production of a mixture of halogenated carbon compounds from a source of electron-rich carbon compounds, such as biomass. The present disclosure also relates to methods of reducing methane emissions in ruminants using the halogenated carbon compounds.BACKGROUND

[0003] Climate change is an increasing problem creating long-term shifts in temperatures and weather patterns. Greenhouse gas emissions from anthropogenic sources have been the main driver of climate change over the last 200 years. These sources are mainly due to energy, industry, transport, buildings, agriculture and land use. Climate change is resulting in increases in global temperatures, as well as increases in the length and severity of droughts, fires, and storms.

[0004] One of the drivers of climate change are greenhouse gas emissions from agriculture. With the global population due to increase to 9 billion by 2050, agricultural production must increase and therefore greenhouse gas emissions will continue to rise. Greenhouse gas emissions from animal agriculture are estimated to represent 14.5 percent of all anthropogenic greenhouse gas emissions.

[0005] There is mounting pressure to reduce herd sizes in an attempt to reduce greenhouse gas emissions from animal agriculture. However, animal products contain protein, fats, vitamins and minerals in the least allergenic and most bioavailable forms for human consumption. As such, alternative methods are required for reducing greenhouse gas emissions from animal agriculture. Some proposed methods include increasing production through herd health and breeding practices, feeding cattle based on nutrient needs, composting of manure, including additives in manure to suppress unwanted biological activity, as well as using manure storage covers and anaerobic digesters to produce methane that is captured and used to replace fossil fuels.

[0006] Recently, it was discovered that supplementing the diet of ruminant animals with the red seaweed species Asparagopsis taxiformis reduced methane emissions. Red seaweed is difficult to cultivate andwild populations are limited. There is thus a need to provide improved methods for reducing methane emissions from ruminants.SUMMARY

[0007] The present inventors have surprisingly found that, when a plurality of electron-rich carbon compounds, which may be present in a source such as biomass, is reacted with a source of hypohalite, the product, which is a plurality of halogenated carbon compounds, is particularly effective at reducing methane emissions in ruminants.

[0008] In a first aspect, there is provided a method of producing a plurality of halogenated carbon compounds, the method comprising contacting a source of electron-rich carbon compounds with a composition comprising hypohalite ions under conditions and for a time sufficient to produce the plurality of halogenated carbon compounds.

[0009] The source of electron-rich carbon compounds may be an organic material. The organic material may be biomass. The biomass may be a microorganism, a plant material, or a combination thereof.

[0010] The electron-rich carbon compounds may comprise a ketone. The ketone may be a methyl ketone. The methyl ketone may have the structure:wherein R1is selected from the group consisting of H, halogen, NO2, CN, NH2, optionally substituted Ci- Ci2alkyl, optionally substituted C2-Ci2alkenyl, optionally substituted C2-Ci2alkynyl, optionally substituted C2-Ci2heteroalkyl, optionally substituted C3-Ci2cycloalkyl, optionally substituted C2-Ci2heterocycloalkyl, optionally substituted C2-Ci2heterocycloalkenyl, optionally substituted C(,-C is aryl, optionally substituted Ci-Cisheteroaryl, optionally substituted Ci-Ci2alkyloxy, optionally substituted C2-Ci2alkenyloxy, optionally substituted C2-Ci2alkynyloxy, optionally substituted C2-Ci2heteroalkyloxy, optionally substituted Cs-C cycloalkyloxy, optionally substituted Cs-C cycloalkenyloxy, optionally substituted Ci- Ci2heterocycloalkyloxy, optionally substituted C2-Ci2heterocycloalkenyloxy, optionally substituted C - Cisaryloxy, optionally substituted Ci-Cisheteroaryloxy, optionally substituted Ci-C alkylamino, SR2, SO3H, SO2NH2, SO2R2, SONH2, SOR2, COR2, COOH, COOR2, CON R2R3, NR2COR3, NR2COOR3, NR2SO2R3, N R2CON R2R3, N R2R3, and acyl; wherein each R2andR3is independently selected from the group consisting of: H, optionally substituted Ci-Ci2alkyl, optionally substituted C2-Ci2alkenyl, optionally substituted C2-Ci2alkynyl, optionally substituted C2-Ci2heteroalkyl, optionally substituted C3-Ci2cycloalkyl, optionally substituted Cs-C cycloalkenyl, optionally substituted CT-C hctcrocycloalkyl, optionally substituted C2-C hctcrocycloalkcnyl, optionally substituted Ce-C-isaryl, and optionally substituted Ci-Cisheteroaryl.

[0011] The halogenated carbon compounds may comprise haloform. The haloform may be bromoform.

[0012] The amount of hypohalite ions in the composition may be at least between about 0.005 mol and about 0.05 mol per gram of the source.

[0013] The composition may be a solution. The solution may be an aqueous solution. The hypohalite ions may be hypobromite ions or hypochlorite ions, or a combination thereof.

[0014] The method may further comprise extracting one or more of the halogenated carbon compounds after producing the plurality of halogenated carbon compounds.

[0015] In a second aspect, there is provided an animal supplement comprising the plurality of halogenated carbon compounds produced by the method of the first aspect, or an extract thereof.

[0016] In a third aspect, there is provided an animal feed comprising the supplement or extract of the second aspect.

[0017] In a fourth aspect, there is provided a method of reducing methane emissions of an animal, comprising administering an effective amount of the supplement of the second aspect or the animal feed of the third aspect to the animal. The animal may be a ruminant animal. The effective amount may be about 0.01% to about 5% dry weight of the animal's diet.

[0018] In a fifth aspect, there is provided a composition for producing a plurality of halogenated carbon compounds, the composition comprising:- a source of electron-rich carbon compounds; and- a composition comprising hypohalite ions.BRIEF DESCRIPTION OF DRAWINGS

[0019] Embodiments of the present disclosure will be discussed with reference to the accompanying drawings wherein:

[0020] Figure 1 shows the ratio of amount of hypohalite ions to mass of biomass for different types of biomass: la yeast; lb oats; 1c wheat; Id corn; le barley.

[0021] Figure 2 shows the amount of bromoform produced per gram of biomass using increasing amounts of hypohalite ions (“bromination solution”): 2a yeast; 2b oat; 2c wheat; 2d corn; 2e barley.

[0022] Figure 3 shows bromoform production from different types of biomass: barley, oats, wheat, corn, and yeast were treated with hypobromite to produce (among other halogenated compounds) bromoform;

[0023] Figure 4 shows that variable bromination conditions with normalised bromoform levels cause variable levels of methanogenesis inhibition: liquid corn extract samples treated with two solutions of different hypobromite concentrations (BSB 25 mM and BSA 6.25 mM) were normalised to different bromoform concentrations via dilution and used to treat in vitro rumen cultures alongside no-treatment controls (methane was quantified using GC-MS after 24 hours of in vitro rumen incubation);

[0024] Figure 5 shows that biomass treated with hypobromite inhibits in vitro rumen methane emissions to a greater extent than bromoform-only biomass: brominated and non-brominated liquid corn extracts with identical bromoform concentrations were used to treat in vitro rumens (methane was quantified using GC-MS after 24 hours of in vitro rumen incubation);

[0025] Figure 6 shows that differentially brominated biomass normalised by bromoform concentration shows differential methanogenesis inhibition levels: different biomasses were treated with different concentrations of hypobromite, and the resulting products were used to treat in vitro rumen cultures (methane was measured using GC-MS after 24 hours of in vitro rumen incubation);

[0026] Figure 7 shows a reduction in methane emissions from ruminants consuming varying amounts of brominated biomass, where: T1 represents 0 g of brominated biomass per kg of dry matter intake (DMI); T2 represents 1.15 g of brominated biomass per kg of dry matter intake (DMI); T3 represents 2.30 g of brominated biomass per kg of dry matter intake (DMI); and T4 represents 4.59 g of brominated biomass per kg of dry matter intake (DMI).DESCRIPTION OF EMBODIMENTSDEFINITIONS

[0027] The following definitions are provided for specific terms which are used in the following written description.

[0028] As used in the specification and claims, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise.

[0029] As used herein, the term "comprising" is intended to mean that the compositions and methods include the recited features, but do not exclude other features. "Consisting essentially of" shall mean excluding other features of any essential significance to the combination. Thus, compositions consisting essentially of a plurality of halogenated carbon compounds would not exclude trace contaminants from the preparation, isolation and purification method, or pharmaceutically acceptable carriers. As it relates to methods, “consisting essentially of’ shall mean excluding other steps of any essential significance to the combination. "Consisting of" shall mean excluding more than trace elements of other ingredients and substantial method steps. Embodiments defined by each of these transition terms are within the scope of this invention.

[0030] The term "about" or "approximately" means within an acceptable range for the particular value as determined by the person skilled in the art, which will also depend, at least in part, on how the value is measured or determined, e.g., the limitations of the measurement system.

[0031] Where a range of values is provided, it is understood that each intervening value, between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either of those included limits are also included in the invention.

[0032] Statistical analysis of the properties described herein may be carried out by standard tests, for example, t-tests, ANOVA, or Chi squared tests. Typically, statistical significance will be measured to a level of p=0.05 (5%), more preferably p=0.01, p=0.001, p=0.0001, p=0.000001

[0033] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by the person skilled in the art to which this invention belongs. All publications mentioned herein are incorporated by reference for the purpose of describing and disclosing devices, formulations and methodologies that may be used in connection with the presently described invention.

[0034] The present inventors' earlier research resulted in WO2020243792A1, directed to bromoform production using yeast expressing vanadium-dependent bromoperoxidase (vBPO). The significance of producing bromoform is that it can interfere directly with the methanogenesis pathway in methanogens by inhibiting a cobamide-dependent methyltransferase. Continued research showed that by following theExample in WO2020243792A1, the results could not be reproduced. Briefly, yeast constitutively expressing vBPO did not produce bromoform in vivo.

[0035] The present inventors conducted further research into the halogenation of biomass and found that, under the right conditions, yeast expressing vBPO was able to produce bromoform in vivo. Surprisingly, the inventors found that the halogenation of biomass to produce a cocktail of halogenated carbon compounds (including bromoform) can also be carried out by using hypohalite ions in the absence of vBPO.

[0036] Accordingly, in a first aspect, there is provided a method of producing a plurality of halogenated carbon compounds, the method comprising contacting a source of electron-rich carbon compounds with a composition comprising hypohalite ions under conditions and for a time sufficient to produce the plurality of halogenated carbon compounds.

[0037] Past research has focussed on the production of bromoform and its application in reducing methane emissions in animals. The present inventors have discovered that the method of the first aspect may produce a cocktail of halogenated products. It has been found that the cocktail is effective for reducing methane emissions and that the concentration of bromoform alone is not indicative of the effect on reducing methane emissions. The inventors have recognised that the production of the cocktail of halogenated products occurs by reaction of a plurality of electron-rich carbon compounds, which may be present in organic materials (such as biomass), with hypohalite ions.

[0038] This method of halogenating a source of electron -rich carbon compounds is also a simpler method than methods involving the use of haloperoxidases, which require particular conditions and the presence of further additional reactants, such as vanadium ions. Accordingly, there is provided a method of producing a plurality of halogenated carbon compounds, the method comprising contacting a source of electron-rich carbon compounds with a composition comprising hypohalite ions in the absence of a haloperoxidase under conditions and for a time sufficient to produce the plurality of halogenated carbon compounds. The haloperoxidase may be a vanadium-dependent haloperoxidase. The vanadium-dependent haloperoxidase may be vanadium-dependent bromoperoxidase, vanadium-dependent chloroperoxidase or vanadium-dependent iodoperoxidase, or any combination thereof.

[0039] There is also provided a method of producing a plurality of halogenated carbon compounds, the method comprising contacting a source of electron-rich carbon compounds with a composition consisting essentially of hypohalite ions under conditions and for a time sufficient to produce the plurality of halogenated carbon compounds.

[0040] There is also provided a method of producing a plurality of halogenated carbon compounds, the method consisting essentially of contacting a source of electron-rich carbon compounds with a composition comprising hypohalite ions under conditions and for a time sufficient to produce the plurality of halogenated carbon compounds.

[0041] There is also provided a method of producing a plurality of halogenated carbon compounds, the method consisting essentially of contacting a source of electron-rich carbon compounds with a composition consisting essentially of hypohalite ions under conditions and for a time sufficient to produce the plurality of halogenated carbon compounds.

[0042] The present inventors have also surprisingly found that a significant amount of bromoform is generated from a smaller amount of electron-rich carbon compound-bearing organic material (e.g., biomass) using the method of the present invention, compared to other methods for producing bromoform. The present inventors also postulate that the product of the method of the present invention, comprising a cocktail of halogenated carbon compounds, may avoid the rumen adaptation that has been reported to occur when single pure methanogenesis inhibitors (like bromochloromethane and 3- nitrooxypropanol) are used - the following publications report a decrease in methanogenesis inhibition over time: Patra, A. et al. (2017) Journal of Animal Science and Biotechnology, 8(13); Tomkins, N. W. et al. (2009) Animal Production Science, 49:1053-1058; McGinn, S. M. et al. (2019) Journal of Environmental Quality, 48:1454-1461.

[0043] The term "electron-rich carbon compound" refers to a compound that includes one or more groups that are susceptible to halogenation by the action of hypohalite ions. Examples of suitable compounds include those containing ketone and / or alkene groups. Organic materials, such as biomass, are composed of a complex mixture of carbon compounds, including compounds comprising alkene and / or ketone groups. Therefore, the source of electron-rich carbon compounds may be an organic material, such as biomass. The biomass may be a microorganism, plant material or a combination thereof. The microorganism may be one or more species of fungus, bacteria or archaea. For example, the microorganism may be a yeast. The plant material may be derived from plant shoot, root or seed, plant waste, biomass feedstocks, crop residue, algae, wood processing residue, corn, rice, wheat, oats, triticale, barley, sorghum, millet, rye, farro, quinoa, soybean, beets, buckwheat, bulgur, amaranth, kamut, spelt, freekeh, teff, fonio, einkorn, sunflower, flaxseed, sesame seed, rapeseed, or cottonseed. Other biomass types could be determined by the person skilled in the art.

[0044] The electron-rich carbon compounds may comprise a ketone. The ketone may be a methyl ketone. The methyl ketone may have the structure:wherein R1is selected from the group consisting of H, halogen, NO2, CN, NH2, optionally substituted Ci-C alkyl, optionally substituted C2-Ci2alkenyl, optionally substituted C2-Ci2alkynyl, optionally substituted C2-Ci2heteroalkyl, optionally substituted Cs-C cycloalkyl, optionally substituted C3-Ci2cycloalkenyl, optionally substituted C2-Ci2heterocycloalkyl, optionally substituted C2- Ci2heterocycloalkenyl, optionally substituted Ce-Cis aryl, optionally substituted Ci-Cisheteroaryl, optionally substituted Ci-Ci2alkyloxy, optionally substituted C2-Ci2alkenyloxy, optionally substituted C2- Ci2alkynyloxy, optionally substituted C2-Ci2heteroalkyloxy, optionally substituted Cs-C cycloalkyloxy, optionally substituted C3-Ci2cycloalkenyloxy, optionally substituted Ci-Ci2heterocycloalkyloxy, optionally substituted C2-Ci2heterocycloalkenyloxy, optionally substituted Ce-Cisaryloxy, optionally substituted Ci-Cisheteroaryloxy, optionally substituted Ci-C alkylamino, SR2, SO3H, SO2NH2, SO2R2, SONH2, SOR2, COR2, COOH, COOR2, CON R2R3, NR2COR3, NR2COOR3, NR2SO2R3, N R2CON R2R3, N R2R3, and acyl; wherein each R2andR3is independently selected from the group consisting of: H, optionally substituted Ci-Ci2alkyl, optionally substituted C2-Ci2alkenyl, optionally substituted C2- Ci2alkynyl, optionally substituted C2-Ci2heteroalkyl, optionally substituted C3-Ci2cycloalkyl, optionally substituted Cs-C cycloalkenyl, optionally substituted C2-Ci2heterocycloalkyl, optionally substituted C2- Ci2heterocycloalkenyl, optionally substituted Ce-C-isaryl, and optionally substituted Ci-Cisheteroaryl.

[0045] The term "optionally substituted" as used throughout the specification denotes that the group may or may not be further substituted or fused (to form a condensed polycyclic system), with one or more nonhydrogen substituent groups. In certain embodiments the substituent groups are one or more groups independently selected from the group consisting of halogen, =0, =S, -CN, -NO2, -CF3, -OCF3, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, heteroarylalkyl, arylalkyl, cycloalkylalkenyl, heterocycloalkylalkenyl, arylalkenyl, heteroarylalkenyl, cycloalkylheteroalkyl, heterocycloalkylheteroalkyl, arylheteroalkyl, heteroarylheteroalkyl, hydroxy, hydroxyalkyl, alkyloxy, alkyloxyalkyl, alkyloxycycloalkyl, alkyloxyheterocycloalkyl, alkyloxyaryl, alkyloxyheteroaryl, alkyloxycarbonyl, alkylaminocarbonyl, alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy, phenoxy, benzyloxy, heteroaryloxy, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, alkylsulfinyl, arylsulfinyl, aminosulfinylaminoalkyl, -C(=0)0H, -C(=O)Ra, -C(=O)ORa, C(=0)NRaRb, C(=N0H)Ra, C(=NRa)NRbRc, NRaRb, NRaC(=0)Rb, NRaC(=0)0Rb, NRaC(=0)NRbRc, N RaC(= N Rb) N R°Rd, NRaSO2Rb,-SRa, SO2NRaRb, -ORa, 0C(=0)NRaRb, OC(=O)Raand acyl, wherein Ra, Rb, Rcand Rdareeach independently selected from the group consisting of H, Ci-C alkyl, Ci-C haloalkyl, C2-Ci2alkenyl, C2-Ci2alkynyl, C2-C10 heteroalkyl, C3-Ci2cycloalkyl, C3-C i2cycloalkenyl, C2-Ci2heterocycloalkyl, C2- Ci2heterocycloalkenyl, Ce-Cisaryl, C2-Cisheteroaryl, and acyl, or any two or more of Ra, Rb, Rcand Rd, when taken together with the atoms to which they are attached form a heterocyclic ring system with 3 to 12 ring atoms.

[0046] In the definitions of a number of substituents below it is stated that "the group may be a terminal group or a bridging group". This is intended to signify that the use of the term is intended to encompass the situation where the group is a linker between two other portions of the molecule as well as where it is a terminal moiety. Using the term alkyl as an example, some publications would use the term "alkylene" for a bridging group and hence in these other publications there is a distinction between the terms "alkyl" (terminal group) and "alkylene" (bridging group). In the present application no such distinction is made and most groups may be either a bridging group or a terminal group.

[0047] "Acyl" means an R-C(=O)- group in which the R group may be an alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group as defined herein. Examples of acyl include acetyl and benzoyl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the carbonyl carbon.

[0048] "Acylamino" means an R-C(=O)-NH- group in which the R group may be an alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the nitrogen atom.

[0049] "Alkenyl" as a group or part of a group denotes an aliphatic hydrocarbon group containing at least one carbon-carbon double bond and which may be straight or branched preferably having 2-12 carbon atoms, more preferably 2-10 carbon atoms, most preferably 2-6 carbon atoms, in the normal chain. The group may contain a plurality of double bonds in the normal chain and the orientation about each is independently E or Z. Exemplary alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl and nonenyl. The group may be a terminal group or a bridging group.

[0050] "Alkenyloxy" refers to an alkenyl-O- group in which alkenyl is as defined herein. Preferred alkenyloxy groups are C2- C alkenyloxy groups. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the oxygen atom.

[0051] "Alkyl" as a group or part of a group refers to a straight or branched aliphatic hydrocarbon group, preferably a C1-C-12 alkyl, more preferably a C1-C10 alkyl, most preferably Ci-Ce alkyl unless otherwisenoted. Examples of suitable straight and branched Ci-Ce alkyl substituents include methyl, ethyl, n- propyl, 2-propyl, n-butyl, sec-butyl, t-butyl, hexyl, and the like. The group may be a terminal group or a bridging group.

[0052] "Alkylamino" includes both mono-alkylamino and dialkylamino, unless specified. "Monoalkylamino" means an Alkyl-NH- group, in which alkyl is as defined herein. "Dialkylamino" means a (alkyl)2N- group, in which each alkyl may be the same or different and are each as defined herein for alkyl. The alkyl group is preferably a Ci-Ce alkyl group. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the nitrogen atom.

[0053] "Alkylaminocarbonyl" refers to a group of the formula (Alkyl)x(H)yNC(=O)- in which alkyl is as defined herein, x is 1 or 2, and the sum of X+Y =2. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the carbonyl carbon.

[0054] "Alkyloxy" refers to an alkyl-O- group in which alkyl is as defined herein. Preferably the alkyloxy is a C i-CTalkyloxy. Examples include, but are not limited to, methoxy and ethoxy. The group may be a terminal group or a bridging group.

[0055] "Alkyloxyalkyl" refers to an alkyloxy-alkyl- group in which the alkyloxy and alkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkyl group.

[0056] "Alkyloxyaryl" refers to an alkyloxy-aryl- group in which the alkyloxy and aryl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the aryl group.

[0057] "Alkyloxycarbonyl" refers to an alkyl-O-C(=O)- group in which alkyl is as defined herein. The alkyl group is preferably a Ci-Ce alkyl group. Examples include, but are not limited to, methoxycarbonyl and ethoxycarbonyl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the carbonyl carbon.

[0058] "Alkyloxycycloalkyl" refers to an alkyloxy-cycloalkyl- group in which the alkyloxy and cycloalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the cycloalkyl group.

[0059] "Alkyloxyheteroaryl" refers to an alkyloxy-heteroaryl- group in which the alkyloxy and heteroaryl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the heteroaryl group.

[0060] "Alkyloxyheterocycloalkyl" refers to an alkyloxy-heterocycloalkyl- group in which the alkyloxy and heterocycloalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the heterocycloalkyl group.

[0061] "Alkylsulfinyl" means an alkyl-S-(=O)- group in which alkyl is as defined herein. The alkyl group is preferably a Ci-Ce alkyl group. Exemplary alkylsulfinyl groups include, but not limited to, methylsulfinyl and ethylsulfinyl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the sulfur atom.

[0062] "Alkylsulfonyl" refers to an alkyl-S(=O)2- group in which alkyl is as defined above. The alkyl group is preferably a Ci-Ce alkyl group. Examples include, but not limited to methylsulfonyl and ethylsulfonyl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the sulfur atom.

[0063] "Alkynyl" as a group or part of a group means an aliphatic hydrocarbon group containing a carbon-carbon triple bond and which may be straight or branched preferably having from 2-12 carbon atoms, more preferably 2-10 carbon atoms, more preferably 2-6 carbon atoms in the normal chain. Exemplary structures include, but are not limited to, ethynyl and propynyl. The group may be a terminal group or a bridging group.

[0064] "Alkynyloxy" refers to an alkynyl-O- group in which alkynyl is as defined herein. Preferred alkynyloxy groups are C2-C6 alkynyloxy groups. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the oxygen atom.

[0065] “Amino” means an NH2 group if it is terminal, and NH if it is a bridging group.

[0066] "Aminoalkyl" means an NlE-alkyl- group in which the alkyl group is as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkyl group.

[0067] "Aminosulfonyl" means an NH2-S(=O)2- group. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the sulfur atom.

[0068] "Aryl" as a group or part of a group denotes (i) an optionally substituted monocyclic, or fused polycyclic, aromatic carbocycle (ring structure having ring atoms that are all carbon) preferably having from 5 to 12 atoms per ring. Examples of aryl groups include phenyl, naphthyl, and the like; (ii) an optionally substituted partially saturated bicyclic aromatic carbocyclic moiety in which a phenyl and a Cs C7 cycloalkyl or C5-C7 cycloalkenyl group are fused together to form a cyclic structure, such as tetrahydronaphthyl, indenyl or indanyl. The group may be a terminal group or a bridging group. Typically an aryl group is a C(,-C ix aryl group.

[0069] "Arylalkenyl" means an aryl-alkenyl- group in which the aryl and alkenyl are as defined herein. Exemplary arylalkenyl groups include phenylallyl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkenyl group.

[0070] "Arylalkyl" means an aryl-alkyl- group in which the aryl and alkyl moieties are as defined herein. Preferred arylalkyl groups contain a Ci-Ce alkyl group. Exemplary arylalkyl groups include benzyl, phenethyl, 1 -naphthalenemethyl and 2-naphthalenemethyl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkyl group.

[0071] "Arylalkyloxy" refers to an aryl-alkyl-O- group in which the alkyl and aryl are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the oxygen atom.

[0072] "Arylamino" includes both mono-arylamino and di-arylamino unless specified. Mono-arylamino means a group of formula arylNH-, in which aryl is as defined herein, di-arylamino means a group of formula ('aryl pN- where each aryl may be the same or different and are each as defined herein for aryl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the nitrogen atom.

[0073] "Arylheteroalkyl" means an aryl-heteroalkyl- group in which the aryl and heteroalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the heteroalkyl group.

[0074] "Aryloxy" refers to an aryl-O- group in which the aryl is as defined herein. Preferably the aryloxy is a Ce-Cisaryloxy, more preferably a Ce-Cwaryloxy. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the oxygen atom.

[0075] "Arylsulfinyl" means an aryl-S(=O)- group in which the aryl group is as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the sulfur atom.

[0076] "Arylsulfonyl" means an aryl-S(=O)2- group in which the aryl group is as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the sulfur atom.

[0077] A "bond" is a linkage between atoms in a compound or molecule. The bond may be a single bond, a double bond, or a triple bond.

[0078] "Cycloalkenyl" means a non-aromatic monocyclic or multicyclic ring system containing at least one carbon-carbon double bond and preferably having from 5-10 carbon atoms per ring. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl or cycloheptenyl. The cycloalkenyl group may be substituted by one or more substituent groups. A cycloalkenyl group typically is a C3-C12 alkenyl group. The group may be a terminal group or a bridging group.

[0079] "Cycloalkyl" refers to a saturated monocyclic or fused or spiro polycyclic, carbocycle preferably containing from 3 to 9 carbons per ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like, unless otherwise specified. It includes monocyclic systems such as cyclopropyl and cyclohexyl, bicyclic systems such as decalin, and polycyclic systems such as adamantane. A cycloalkyl group typically is a C3-C12 alkyl group. The group may be a terminal group or a bridging group.

[0080] "Cycloalkylalkyl" means a cycloalkyl-alkyl- group in which the cycloalkyl and alkyl moieties are as defined herein. Exemplary monocycloalkylalkyl groups include cyclopropylmethyl, cyclopentylmethyl, cyclohexylmethyl and cycloheptylmethyl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkyl group.

[0081] "Cycloalkylalkenyl" means a cycloalkyl-alkenyl- group in which the cycloalkyl and alkenyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkenyl group.

[0082] "Cycloalkylheteroalkyl" means a cycloalkyl-heteroalkyl- group in which the cycloalkyl and heteroalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the heteroalkyl group.

[0083] "Cycloalkyloxy" refers to a cycloalkyl-O- group in which cycloalkyl is as defined herein. Preferably the cycloalkyloxy is a Ci-Ce cycloalkyloxy. Examples include, but are not limited to, cyclopropanoxy and cyclobutanoxy. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the oxygen atom.

[0084] "Cycloalkenyloxy" refers to a cycloalkenyl-O- group in which the cycloalkenyl is as defined herein. Preferably the cycloalkenyloxy is a CT-Cs cycloalkenyloxy. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the oxygen atom.

[0085] "Haloalkyl" refers to an alkyl group as defined herein in which one or more of the hydrogen atoms has been replaced with a halogen atom selected from the group consisting of fluorine, chlorine, bromine and iodine. A haloalkyl group typically has the formula CnH(2n+i-m) m wherein each X is independently selected from the group consisting of F, CI, Br and I. In groups of this type n is typically from 1 to 10, more preferably from 1 to 6, most preferably 1 to 3. m is typically 1 to 6, more preferably 1 to 3. Examples of haloalkyl include fluoromethyl, difluoromethyl and trifluoromethyl.

[0086] "Haloalkenyl" refers to an alkenyl group as defined herein in which one or more of the hydrogen atoms has been replaced with a halogen atom independently selected from the group consisting of F, CI, Br and I.

[0087] "Haloalkynyl" refers to an alkynyl group as defined herein in which one or more of the hydrogen atoms has been replaced with a halogen atom independently selected from the group consisting of F, CI, Br and I.

[0088] "Halogen" refers to chlorine, fluorine, bromine or iodine.

[0089] "Heteroalkyl" refers to a straight- or branched-chain alkyl group preferably having from 2 to 12 carbons, more preferably 2 to 6 carbons in the chain, in which one or more of the carbon atoms (and any associated hydrogen atoms) are each independently replaced by a heteroatomic group selected from S, O, P and NR' where R' is selected from the group consisting of H, optionally substituted Ci-Ci2alkyl, optionally substituted CT-C cycloalkyl, optionally substituted Ce-C-isaryl, and optionally substituted Ci- Cisheteroaryl. Exemplary heteroalkyls include alkyl ethers, secondary and tertiary alkyl amines, amides, alkyl sulfides, and the like. Examples of heteroalkyl also include hydroxyC i -Chalky 1, Ci-Cealkyloxy Ci- CTalkyl, amino Ci-Cealkyl, Ci-CealkylaminoCi-Cealkyl, and di(Ci-C6alkyl)amino Ci-Cealkyl. The group may be a terminal group or a bridging group.

[0090] "Heteroalkyloxy" refers to a heteroalkyl-O- group in which heteroalkyl is as defined herein. Preferably the heteroalkyloxy is a Cz-Ceheteroalkyloxy. The group may be a terminal group or a bridging group.

[0091] "Heteroaryl" either alone or part of a group refers to groups containing an aromatic ring (preferably a 5 or 6 membered aromatic ring) having one or more heteroatoms as ring atoms in the aromatic ring with the remainder of the ring atoms being carbon atoms. Suitable heteroatoms include nitrogen, oxygen and sulphur. Examples of heteroaryl include thiophene, benzothiophene, benzofuran, benzimidazole, benzoxazole, benzothiazole, benzisothiazole, naphtho[2,3-b]thiophene, furan, isoindolizine, xantholene, phenoxatine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine,l ,3,5-triazene, tetrazole, indole, isoindole, 1 H-indazole, benzotriazole, purine, quinoline, isoquinoline, phthalazine, naphthyridine, quinoxaline, cinnoline, carbazole, phenanthridine, acridine, phenazine, thiazole, isothiazole, phenothiazine, oxazole, isooxazole, 1 ,2,3-oxadiazole, 1 ,2,4-oxadiazole, 1 ,2, 3 -triazole, 1 ,2,4-triazole, 1 ,2,4-thiadiazole, 1 ,3,4-thiadiazole, furazane, phenoxazine, 2-, 3- or 4- pyridyl, 2-, 3-, 4-, 5-, or 8- quinolyl, 1-, 3-, 4-, or 5-isoquinolinyl 1-, 2-, or 3- indolyl, and 2-, or 3-thienyl. A heteroaryl group is typically a Cl-C-18 heteroaryl group. The group may be a terminal group or a bridging group.

[0092] "Heteroarylalkyl" means a heteroaryl-alkyl group in which the heteroaryl and alkyl moieties are as defined herein. Preferred heteroarylalkyl groups contain a lower alkyl moiety. Exemplary heteroarylalkyl groups include pyridylmethyl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkyl group.

[0093] "Heteroarylalkenyl" means a heteroaryl-alkenyl- group in which the heteroaryl and alkenyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkenyl group.

[0094] "Heteroarylheteroalkyl" means a heteroaryl-heteroalkyl- group in which the heteroaryl and heteroalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the heteroalkyl group.

[0095] "Heteroaryloxy" refers to a heteroaryl-O- group in which the heteroaryl is as defined herein. Preferably the heteroaryloxy is a Ci Cis heteroaryloxy. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the oxygen atom.

[0096] "Heterocyclic" refers to saturated, partially unsaturated or fully unsaturated monocyclic, bicyclic or polycyclic ring system containing at least one heteroatom selected from the group consisting ofnitrogen, sulfur and oxygen as a ring atom. Examples of heterocyclic moieties include heterocycloalkyl, heterocycloalkenyl and heteroaryl.

[0097] "Heterocycloalkenyl" refers to a heterocycloalkyl group as defined herein but containing at least one double bond. A heterocycloalkenyl group typically is a C2-C12 heterocycloalkenyl group. The group may be a terminal group or a bridging group.

[0098] "Heterocycloalkyl" refers to a saturated monocyclic, bicyclic, or polycyclic ring containing at least one heteroatom selected from nitrogen, sulfur, oxygen, preferably from 1 to 3 heteroatoms in at least one ring. Each ring is preferably from 3 to 10 membered, more preferably 4 to 7 membered. Examples of suitable heterocycloalkyl substituents include pyrrolidyl, tetrahydrofuryl, tetrahydrothiofuranyl, piperidyl, piperazyl, tetrahydropyranyl, morphilino, 1 ,3-diazapane, 1 ,4-diazapane, 1 ,4- oxazepane, and 1 ,4- oxathiapane. A heterocycloalkyl group typically is a C2-C12 heterocycloalkyl group. The group may be a terminal group or a bridging group.

[0099] "Heterocycloalkylalkyl" refers to a heterocycloalkyl-alkyl- group in which the heterocycloalkyl and alkyl moieties are as defined herein. Exemplary heterocycloalkylalkyl groups include (2- tetrahydrofuryl)methyl, (2 -tetrahydrothiofuranyl) methyl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkyl group.

[0100] "Heterocycloalkylalkenyl" refers to a heterocycloalkyl-alkenyl- group in which the heterocycloalkyl and alkenyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkenyl group.

[0101] "Heterocycloalkylheteroalkyl" means a heterocycloalkyl-heteroalkyl- group in which the heterocycloalkyl and heteroalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the heteroalkyl group.

[0102] "Heterocycloalkyloxy" refers to a heterocycloalkyl-O- group in which the heterocycloalkyl is as defined herein. Preferably the heterocycloalkyloxy is a Ci- Ceheterocycloalkyloxy. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the oxygen atom.

[0103] "Heterocycloalkenyloxy" refers to a heterocycloalkenyl-O- group in which heterocycloalkenyl is as defined herein. Preferably the heterocycloalkenyloxy is a Ci- Cheterocycloalkenyloxy. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the oxygen atom.

[0104] “Hydroxy” refers to an OH group.

[0105] "Hydroxyalkyl" refers to an alkyl group as defined herein in which one or more of the hydrogen atoms has been replaced with an OH group. A hydroxyalkyl group typically has the formula CnH(2n+i x)(OH)x. In groups of this type n is typically from 1 to 10, more preferably from 1 to 6, most preferably 1 to 3. x is typically 1 to 6, more preferably 1 to 3.

[0106] "Sulfinyl" means an R-S(=O)- group in which the R group may be OH, alkyl, cycloalkyl, heterocycloalkyl; aryl or heteroaryl group as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the sulfur atom.

[0107] "Sulfinylamino" means an R-S(=O)-NH- group in which the R group may be OH, alkyl, cycloalkyl, heterocycloalkyl; aryl or heteroaryl group as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the nitrogen atom.

[0108] "Sulfonyl" means an R-S(=O)2- group in which the R group may be OH, alkyl, cycloalkyl, heterocycloalkyl; aryl or heteroaryl group as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the sulfur atom.

[0109] "Sulfonylamino" means an R-S(=O)2-NH- group. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the nitrogen atom.

[0110] Some of the compounds of the disclosed embodiments may exist as single stereoisomers, racemates, and / or mixtures of enantiomers and / or diastereomers. All such single stereoisomers, racemates and mixtures thereof, are intended to be within the scope of the subject matter described and claimed. The isomeric forms such as diastereomers, enantiomers, and geometrical isomers can be separated by physical and / or chemical methods known to those skilled in the art.

[0111] The term “hypohalite ion” refers to an oxy anion containing a halogen in the +1 oxidation state or in the -1 oxidation state. Oxy anions with a halogen in the +1 oxidation state include hypoiodite (IO ), hypobromite (BrO ) and hypochlorite (CIO ). Oxy anions with a halogen in the -1 oxidation stateinclude hypofluorite (FO ). Therefore, the hypohalite ions may be hypoiodite ions, hypobromite ions, hypochlorite ions, or hypofluorite ions, or a combination thereof. The hypohalite ions may be hypoiodite ions, hypobromite ions, or hypochlorite ions, or a combination thereof. The hypohalite ions may be hypobromite ions.

[0112] Sources of hypohalite ions include salts, such as sodium hypohalites (e.g., NalO, NaBrO, NaClO and NaFO) and potassium hypohalites (e.g., KIO, KBrO, KC1O and KFO). Therefore, the composition comprising hypohalite ions may be a solution, such as an aqueous solution, comprising a hypohalite salt. The hypohalite salt may be sodium hypohalite, potassium hypohalite, or a combination thereof. The composition comprising hypohalite ions may be bleach. The bleach may be chlorine bleach or bromine bleach, or a combination thereof. Hypohalite ions can also be generated in situ via several different reactions, including (where X is Br, Cl or I): a. X2+ 2OH -> X + XO + H2O; b. X2+ H2O — > HXO + HX (which may be preceded by Cl2+ 2X — > 2C1 + X2); c. X + H2O2XO + H2Oe. 2X + O2+ 2OH 2X0 + H2O; f. O3+ X + H2O O2+ OH + HXO g. Br2R + 2H2O 2HOBr + H2R.

[0113] Sources of hypohalite ions also include organic halogenating compounds, such as 1,3- dibromo-5,5-dimethylhydantoin (DBDMH). In this case, reaction g. above, where H2R is 5,5- dimethylhydantoin, is relevant. The composition comprising hypohalite ions may therefore be a solution, such as an aqueous solution, comprising an organic halogenating compound, such as DBDMH. The composition may also comprise a base, such as sodium hydroxide.

[0114] A person skilled in the art will appreciate that the hypohalite ions for use in the method of the present invention may therefore be generated in situ for the purposes of reacting them with the electron-rich carbon compounds to produce the plurality of halogenated carbon compounds, and that the composition comprising hypohalite ions may therefore also comprise one or more reagents required to generate the hypohalite ions. Suitable reagents in this regard include a halogen (such as bromine,chlorine, fluorine or iodine), a halogen salt (such as a potassium or sodium salt of bromide or chloride), an organic halogenating compound (such as DBDMH), a base (such as hydroxide), a peroxide (such as hydrogen peroxide), oxygen or ozone, or a combination thereof. The composition may consist essentially of hypohalite ions, water or a different solvent, and optionally one or more reagents selected from a halogen (such as bromine, chlorine, fluorine or iodine), a halogen salt (such as a potassium or sodium salts of bromide or chloride), an organic halogenating compound (such as DBDMH), a base (such as hydroxide), a peroxide (such as hydrogen peroxide), oxygen and ozone.

[0115] The method comprises contacting a source of electron-rich carbon compounds with a composition comprising hypohalite ions. The hypohalite ions react with electron-rich carbon compounds by electrophilic attack to produce halogenated carbon compounds. It will be understood by a person skilled in the art that any ketones present may undergo reaction with the hypohalite ions to produce haloform (CHX3, where X is a halogen) according to the well-known haloform reaction, which involves exhaustive halogenation of the acetyl group in the presence of a base. Accordingly, one of the halogenated carbon compounds produced may be haloform (e.g., bromoform). Ideally, the composition will need to comprise hypohalite ions in an amount sufficient to halogenate the electron -rich carbon compounds such that the concentration of haloform produced is sufficient to achieve methane reduction in a ruminant’s stomach when administered (e.g., fed) to a ruminant. For example, a minimum concentration of 5-10 pM haloform (e.g., bromoform) may be required, and a concentration of 5-30 pM may be suitable to achieve methane reduction in a ruminant’s stomach.

[0116] A person skilled in the art will appreciate that the minimum amount of hypohalite ions to be used may depend on the source of electron-rich carbon compounds that is to be contacted with the hypohalite ions. For example, the present inventors have found that different biomass types produce optimum amounts of bromoform when the amounts of hypohalite ions used are as shown in the following table.TABLE 1Moles of NaBrO needed per gram of biomass to achieve an effective final concentration of bromoform in the biomass.

[0117] This is depicted graphically in Figure 1. However, smaller or larger amounts than this could also be used.

[0118] The present inventors have found that it may be possible to use smaller amounts of hypohalite ions (e.g., less than about 0.005 mol per gram of the source, such as about 0.001 mol, about 0.0015 mol, or about 0.002 mol per gram of the source) when the hypohalite ions are generated in situ with pre-treated biomass over an extended period. This can be achieved by, for example, first mixing the source with a base (such as sodium hydroxide) and then adding the one or more reagents required to generate the hypohalite ions to the basic mixture slowly over an extended period, allowing the hypohalite ions to react with the biomass as the hypohalite ions are generated. The inventors postulate that the base is free to react with the biomass initially, possibly altering its effective concentration by the time the reagent is added. Additionally, the base may be reacting with precursors and altering the effective concentration of molecules susceptible to halogenation. As such, the optimum ratio of hypohalite ions to biomass may change as the biomass is altered prior to addition of the source of hypohalite ions.

[0119] Therefore, in another aspect, there is provided a method of producing a plurality of halogenated carbon compounds, the method comprising contacting a source of electron-rich carbon compounds with a base (as defined herein), followed by slow addition of a composition comprising, or consisting essentially of, hypohalite ions (as defined herein), under conditions and for a time sufficient to produce the plurality of halogenated carbon compounds. The base and source of electron-rich carbon compounds may be contacted for about five minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes or longer, including up to about one hour or about two hours, prior to addition of the composition comprising hypohalite ions. The contacting may involve stirring. The slow addition may be carried out by adding the composition in aliquots. The slow additionmay be carried out by adding the composition over the course of about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes or more, up to about an hour, or about two hours, or more.

[0120] Accordingly, the minimum amount of hypohalite ions in the composition may be between about 0.001 mol and about 0.05 mol per gram of the source. The amount of hypohalite ions in the composition may be at least about 0.001 mol to about 0.05 mol per gram of the source. The amount of hypohalite ions in the composition may be at least about 0.001, about 0.002, about 0.003. about 0.004 or about 0.005 mol per gram of the source. A person skilled in the art will also appreciate that the reaction of the electron-rich carbon compounds with increasing amounts of the hypohalite ions should continue to produce more of the halogenated product, until the electron-rich carbon compounds have been exhausted. Figure 2 demonstrates this trend. Accordingly, the maximum amount of hypohalite ions that a person skilled in the art may choose to use will depend on various factors such as the amount of halogenated carbon compounds sought to be produced, and the type of biomass.

[0121] The concentration of electron-rich carbon compounds in the source may be between about 4 and about 20 mM, for example about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM or about 20 mM.

[0122] The composition comprising hypohalite ions may be a solution. The solution may be an organic solution (i.e., comprising one or more organic solvents) or an aqueous solution (i.e., comprising water).

[0123] The contacting may be carried out at a temperature at which the loss of halogenated carbon compounds (through evaporation) is minimised or avoided. The contacting is carried out at a temperature of between about 15 and about 30 °C, between about 16 and about 29 °C, between about 17 and about 28 °C, between about 18 and about 27 °C, between about 19 and about 26 °C, or between about 20 and about 25 °C. The temperature may be about 15 °C, about 16 °C, about 17 °C, about 18 °C, about 19 °C, about 20 °C, about 21 °C, about 22 °C, about 23 °C, about 24 °C, about 25 °C, about 26 °C, about 27 °C, about 28 °C, about 29 °C or about 30 °C. The contacting may be carried out at room temperature. The contacting will occur for a period of time sufficient to produce at least one halogenated compound. In certain embodiments, the time is about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, or between about 1 minute and about 12 hours, about 1 minute and about 6 hours, about 1 minute and about 5 hours, about 1 minutes and about 4 hours, about 1 minute and about 3 hours, about 1 minute and about 2 hours, or about 1 minute and about 1 hour. For larger scale production, the contacting may occur for more than one hour, and may occur for as long as about 12 hours or about 24 hours.

[0124] The halogenation reaction may be quenched once a person skilled in the art has decided that the desired amount of halogenated carbon compounds has been produced. The halogenation reaction may be quenched by addition of a suitable reagent, such as sodium thiosulphate, cyclodextrin (e.g., P- cyclodextrin), and the like.

[0125] The method results in the production of a plurality of halogenated carbon compounds. Where the source of the electron-rich carbon compounds comprises a mixture of electron-rich carbon compounds (as is the case with, for example, biomass), it will be appreciated by a person skilled in the art that a plurality of halogenated carbon compounds will be produced, which plurality is a mixture of structurally diverse halogenated carbon compounds. The halogenated carbon compounds may include one or more of 1 -bromo-heptyl bromide, 1 -bromo-hexyl bromide, dibromochloromethane, bromoform, bromochloroacetate, dibromoacetate, bromochlorodimenone, diiodomethane, chloroiodomethane, bromoiodomethane, dibromomethane, methyl iodide, bromochloromethane, bromodichloromethane, dichloromethane, dibromoiodomethane, trichloroacetaldehyde, bromochloroacetic acid, dibromoacetic acid, 3,3-dibromoacrylic acid, iodoform, tetrabromomethane, tribromomethane, trichloromethane, tetrachloromethane, l,l,3,3-tetrabromopropan-2-ol, tribromomethanol, 1,1,3,3-tetrabromoacetone, 1,3- dichloroacetone, 1 , 1 ,3,3-tetrabromopropan-2-one, 1 ,1 ,3,3-tetrachloropropan-2-one, 1 , 1 ,3-tribromo-3- chloropropan-2-one, 1 , 1 ,3-tribromopropan-2-one, 1 , 1 ,3-trichloropropan-2-one, 1 , 1 ,4,4-tetrabromobut-3- en-2-one, 1 , 1 -dibromo-3,3-dichloropropan-2-one, 1 , 1 -dibromo-3-chloropropan-2-one, 1 , 1 -dibromo-3- iodopropan-2-one, 1 , 1 -dibromopropan-2-one, 1 ,3-dibromo- 1 ,3-dichloropropan-2-one, 1 ,3-dibromo- 1 - chloropropan-2-one, l,3-dibromopropan-2-one, l,3-dichloropropan-2-one, l-bromo-1,3,3- trichloropropan-2-one, 1 -bromo- 1 ,3-dichloropropan-2-one, 1 -bromo-3-chloropropan-2-one, 1 -chloro-3- iodopropan-2-one, 3-bromo-l,l-dichloropropan-2-one, 2,2,2-tribromoacetic acid (ethyl ester), 2,2- dibromoacetic acid, 2,2-dibromoacetic acid (ethyl ester), 2,2-dibromoacetic acid (methyl ester), 2- bromo-2-chloroacetic acid, 2,3-dibromoprop-2-enoic acid (ethyl ester), 3,3-dibromoprop-2-enoic acid, 3,3-dibromoprop-2-enoic acid (ethyl ester), 3,3-dibromoprop-2-enoic acid (methyl ester), 3-bromo-2- iodoprop-2-enoic acid, 3-bromo-3-chloroprop-2-enoic acid (ethyl ester), 3-bromoprop-2-enoic acid (ethyl ester), 2-bromoacetaldehyde, 2,2-dibromoacetaldehyde, fluoroacetate, bromopyrroles, brominated cyclic sesquiterpenes (such as a-snyderol, -snyderol, and y-snyderol), halogenated fatty acids (such as 3- bromo-2-heptanoic acids and 3-bromo-2-nonanoic acids), halogenated oxylipins (e.g., chlorinated egregiachlorides A, B and C, chlorinated eiseniachlorides A, B and C, and iodinated eiseniaiodides A, B), halogenated terpenoids (e.g., diterpenes and triterpenes), halogenated indoles, halogenated acetogenins, halogenated naphthalenes, halogenated furanones, halogenated heptan-2-ones, and halogenated phenols. Given environmental concerns, it may be desirable to produce brominated and iodinated compounds, but not chlorinated compounds.

[0126] The method of the present invention may further comprise extracting one or more of the plurality of halogenated carbon compounds after producing the plurality of halogenated carbon compounds. As would be understood by the person skilled in the art, there are many suitable extraction methods, and the appropriate method may depend on the volatility of the halogenated carbon compounds. Suitable methods include, e.g., solvent extraction (using e.g., dichloromethane, hexane, chloroform, ethyl acetate, acetone, methanol or water), headspace analysis, purge-and-trap collection, direct collection on adsorbent resins, enzyme-assisted extraction, microwave-assisted extraction, supercritical fluid extraction, ultrasound-assisted extraction or combinations thereof. The produced halogenated carbon compounds may be identified by a variety of methods, including, e.g., gas chromatography (GC) with mass spectrometry (MS), flame ionization detection (GC / FID), halogen-sensitive detection (GC / HSD) or electron-capture detection (GC / ECD).

[0127] In a second aspect, there is provided an animal supplement comprising the plurality of halogenated carbon compounds produced by the method of the present disclosure, or an extract thereof. The animal supplement may be in any form that is suitable for feeding to an animal, particularly a ruminant. For example, the supplement may be a liquid (e.g., dissolved or suspended in a carrier, such as water), or a solid. Suitable solid forms include powders (e.g., dried, lyophilised, ground, milled), granulates, coated carriers (e.g., spray-dried granules), and the like. The supplement may take the form of a bolus or medicament. As would be appreciated by a person skilled in the art, an amount or concentration of the plurality of halogenated compounds in a dose will depend on the dosage form. For example, a dose in the form a medicament may be concentrated, whereas a dose in the form of a feed may be dilute. The dose may be calculated as a daily dose. For example, the daily dosage of the plurality of halogenated compounds may be the same, whether administered as, e.g., a medicament or as a feed.

[0128] In certain embodiments when the supplement is in a solid form, the supplement may comprise a coating. Alternatively, the supplement may alternatively be a capsule. The coating may serve various functions, e.g., reducing degradation of the plurality of halogenated compounds, improving storage stability, or providing controlled release, e.g. delayed and / or sustained release. A coated supplement may be provided by first preparing the supplement in a solid form and then coating the composition. Useful coatings comprise at least one of the following: acrylic acid, alginate, betacyclodextrin, cyanoacrylate, ethyl cellulose, gylcerin, glycerol, hard fat, hydroxyethyl methacrylate - oligo(hydroxybutyrate), isobutyleneisoprene 20 copolymers, methacrylamide chitosan, milk powder, N- isopropylacrylamide (NIPAAm), poly(methyl methacrylate) capsulation, oils (e.g. vegetable / seed oils, coconut oil, canola oil), poly(D,L-lactide-co-glycolide) (PLGA), poly(vinyl pyrrolidone) (PVP), polydimethylsiloxane (PDMS), polyisobutylene, polyvinyl acetate, polyisoprene, polyethylene, silica hydrogels, sodium alginate, styrene-butadiene copolymers, vinyl acetate, and combinations thereof. Such coatings can be used individually, or in combination. A liquid supplement may comprise a carrier, suchwater or oil. The plurality of halogenated compounds may be dissolved or suspended in the carrier, as appropriate. In certain embodiments, the oil is an edible oil, as such a vegetable / seed oil (e.g. canola oil).

[0129] In some embodiments, such as when a supplement of the disclosure is in the form of a medicament, the supplement comprises a pharmaceutically acceptable carrier and / or diluent such as water, saline, an emulsion, a gel, a hydrogel, or a solid for pellets. In some embodiments, the supplement comprises one or more excipients. Useful excipients comprise, for example, anti-adherents, binders, density adjusters, water soluble bulking agents, water insoluble bulking agents, coatings, colouring, disintegrants, flavourings, glidants, lubricants, preservatives, sorbents, sweeteners, and polymers. In certain embodiments, the supplements of the disclosure are provided in aqueous solution, in saline solution, as an emulsion, as a gel, as a hydrogel, as a paste, in an oil, as a pellet, or as a powder.

[0130] In some embodiments, a supplement is in the form of a bolus and the supplement comprises conventional excipients for a bolus for ruminants. Useful excipients comprise, for example, iron oxide, zinc oxide, magnesium stearate, hydrogenated fat, or combinations thereof (e.g. all of these). A bolus may optionally include a casing, and / or a coating. Administration of a bolus may be on a regular basis, e.g., monthly, weekly, bi-weekly or as described elsewhere herein.

[0131] A supplement may be suitable for adding to feed prior to consumption. In certain embodiments, the supplement comprises additional feed components. Useful feed components comprise, for example, oats, maize, barley, soya, dried distiller’s grain (DDGs) and / or molasses. In certain embodiments, the supplement is in the form of a pellet or powder. The supplement may be incorporated into a concentrate ration or a feed nut.

[0132] In a third aspect, there is provided an animal feed comprising the supplement or extract of the present disclosure. The animal feed may be in any form that is suitable for feeding to an animal, particularly a ruminant. The animal feed may be in the form of a block (e.g., a lick-block or a fat block), a blend (e.g., with a meal, such as rapeseed meal, flaxseed meal, cottonseed meal, soybean meal or cornmeal), a pellet or a diet supplement (such as a milk supplement), or added to the animal's normal drinking water (either dissolving or remaining suspended in the water). In some embodiments, the feed is suitable for the animal's entire diet.

[0133] In a fourth aspect, there is provided a method of reducing methane emissions of an animal, comprising administering an effective amount of the supplement of the present disclosure, or the animal feed of the present disclosure, to the animal. The animal may be a ruminant animal. The effective amount may be about 0.01% to about 5% dry weight of the animal's diet.

[0134] The animal is preferably a ruminant animal. As the person skilled in the art would appreciate, the present disclosure may be useful in ruminants that are raised for food production. In certain embodiments, the ruminant is of the family Tragulidae, Cervidae, Giraffidae, Antilocapridae, Moschidae or Bovidae. In certain embodiments, the ruminant is a domesticated ruminant, for example, in particular embodiments, the ruminant is a mouse-deer, deer, moose, giraffe and okapi, pronghorn, musk deer, cow, goat, sheep, llama, alpaca, bison, elk, reindeer, yak, camel, buffalo or antelope. In particular embodiments, the ruminant is one or more of a cow or sheep.

[0135] The effective amount may be about 0.01% to about 5% dry weight of the animal's diet. The effective amount may be about 0.02%, about 0.05%, about 0.1%, about 0.5%, about 1%, about 1.5, about 2%, about 2.5% or about 3% dry weight of the animal’s diet. For example, if the animal consumes 10 kg of feed per day, then the effective amount is in the range of about 2 g and 300 g. An amount of the produced plurality of halogenated compounds can be adjusted based on the amount of supplement or feed given to the animal, on the animal’s weight, on the desired rumen concentration of the plurality of halogenated compounds, or on a desired decrease in methane production from the animal. For instance, a particular amount of the plurality of halogenated compounds might be added per kg of feed, to control the amount of the plurality of halogenated compounds administered to the animal. Therefore, in certain embodiments, the methods include steps of (i) mixing the plurality of halogenated compounds with food and (ii) oral administration of the food to the animal.. The person skilled in the art would be able to formulate an appropriate dosage form and regime guided by the published literature, e.g., WO2015109362A2. For example, the administering may be at least about every 12 hours, about every 18 hours, daily, every second day, every third day or every fourth day. The administering may occur every time the animal is fed. The administering the supplement or feed according to the above regimes may occur for a period of at least about 5, about 10, about 25, about 50, about 100, about 250 or about 350 days.

[0136] The amount of methane produced by the animal may be reduced by at least about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, about 15%, or about 10% compared to a reference. In certain embodiments, the reference is the amount of methane produced when animals are not administered an effective amount of the supplement of the disclosure or the animal feed of the disclosure to the animal. As would be understood by the person skilled in the art, an "effective amount" may be a quantity of the supplement or animal feed sufficient to allow improvement, e.g., reduction in the amount of methane production in comparison with a reference or control. The reduction in methane can be measured by conventional means, such as the use of respiration chambers, the sulphur hexafluoride (SF6) tracer technique, breath sampling during milking or feeding, the GreenFeed system, or using a laser methane detector, or using an artificial rumen system as described elsewhere in this disclosure, or as described in Hano, J. Gen. Appl. Microbiol., 39, 35-45,1993.

[0137] In a fifth aspect, there is provided a composition for producing a plurality of halogenated carbon compounds, the composition comprising:- a source of electron-rich carbon compounds; and- a composition comprising hypohalite ions.

[0138] The features of the fifth aspect are to be understood as they are described in the first aspect or elsewhere in this disclosure, unless indicated otherwise by context. The components of the composition of the fifth aspect may be combined as described elsewhere herein to produce the plurality of halogenated carbon compounds, for example, the "contacting" of the first aspect may be equivalent to combining the listed components of the composition of the fifth aspect.EXAMPLES

[0139] Example 1

[0140] Production of bromoform from different biomass types

[0141] Barley, oats, wheat, corn, and yeast were treated with a solution comprising sodium hydroxide and bromine to produce a mixture of halogenated carbon compounds, including bromoform. The hypobromite was therefore generated in situ by the reaction of hydroxide with bromine, as follows:

[0142] Each material was ground using a manually operated Victoria Grain Mill. 1 g of each material was incubated at room temperature without agitation for 1 hr in the presence of different amounts of NaBrO (“brominating solution”) to demonstrate the relationship between the amount of hypobromite and bromoform production, and the maximum amount of hypobromite solution required for each type of biomass within the assay time-frame. The maximum amounts of hypohalite for each material were determined using a “residual bromination assay” where 1 g of material was treated with brominating solution every minute in the presence of phenol red until the formation of bromophenol blue was evident, indicating that there was an excess of brominating solution at that time -point and biomass amount / type. The concentrations of hypobromite used for barley were “0.25” (32 mM), “0.5” (64 mM), and “0.75” (96 mM). For oats, concentrations of “0.25” (55 mM), “0.5” (110 mM), and “0.75” (166 mM) were used. For wheat and corn concentrations of “0.25” (48 mM), “0.5” (48 mM), and “0.75” (72 mM) were used. The reactions were stopped via the addition of 1 M sodium thiosulfate, and bromoform was measured using GC-MS headspace analysis (as discussed below). Bromoform was produced from each biomass typeproportionally to the amount of brominating solution included in the reaction, with oats having the highest bromoform yields under these conditions (see Figure 3).

[0143] GC-MS analysis for quantification of bromoform

[0144] Bromoform concentration was quantified using a GC-MS (Shimadzu Nexis 2030 with TQ-8050 NX). 1 mL of the reaction product was transferred to 20 mL headspace vials. The samples were incubated at 60°C while shaking for 10 min. SPME extraction was performed using a divinylbenzene / PDMS fibre coupled to an AOC-6000 autosampler. The fibre was equilibrated at 200° C for 15 min prior to each injection. Extraction time was 5 min at 60°C. Desorption was performed for 10 min.

[0145] The GC-MS was run in Split mode (1:10) using a SH-I-5Sil MS column, helium carrier gas and a column flow rate of 1.9 mL / min. Bromoform identification was based on target ion m / z 173 and reference ions m / z 171 and m / z 175. Quantification was based on calibration curves from pure bromoform standards prepared from a saturated stock solution in water (12.7 mM). Integration was performed on peak areas of ion m / z 173. The column temperature conditions were 50°C for 1 min followed by an increase to 280°C at a rate of 50°C / min.

[0146] Example 2

[0147] Production of other halogenated carbon compounds that inhibit methanogenesis

[0148] To test whether there were compounds other than bromoform present in biomass treated with hypobromite solution, which compounds inhibited methanogenesis, biomass was treated with two different concentrations of hypobromite in solution (as generated in Example 1) at room temperature for 1 min with no agitation. Brominating Solution B (BSB) contained 100 pL of liquid corn extract and 25 mM of hypobromite in a final volume of 2 mL. Brominating Solution A (BSA) was the same except the hypobromite was present at a concentration of 6.25 mM. To stop the bromination reaction, each solution was incubated for 1 min prior to addition of 5 pL of 5 M sodium thiosulfate. The inhibition testing was carried out as described in the following paragraph. Bromoform levels were quantified via GC-MS as described above, and BSA and BSB were then used to prepare samples with bromoform at either 1 pM, 2.5 pM, or 5 pM by dilution in water.

[0149] Rumen fluid was collected and pooled from several grass-fed fistulated cattle, filtered through muslin cloth and stored at 4 °C for up to 8 days before use in in vitro rumen fermentations. Rumen fluid fermentations were set up in 100 mL glass serum vials containing 4:1 modified in vitro Rumen Fermentation Buffer (RFB) (excluding trypticase): rumen fluid, to a total volume of 25 mL(Goering, H.K. and Van Soest, P.J., 1970. Forage fiber analyses (apparatus, reagents, procedures, and some applications) (No. 379). US Agricultural Research Service). 500 mg of ground Rhodes grass hay (Chloris gayana) was added to the vials along with the RFB and the vials capped with butyl rubber stoppers and aluminium seals and heated to 55 °C to degas oxygen from the buffer liquid. Once vials reached temperature they were flushed with N2 to create anoxia before being cooled and the reductant solution (Goering and Van Soest, 1970) and 1.5 mL of treatments were added to vials via syringe. At this time, treatments prepared with either BSA or BSB (as described above) were added via syringe. Finally, rumen fluid was also added via syringe and vials incubated at 38 °C with shaking at 200 rpm.

[0150] Methane quantification was performed using GC-MS analysis on the same machine as that given above for the quantification of bromoform. After incubation of in vitro rumens with bromination solution, 10 pL of samples were removed from the headspace of the serum bottles and analysed using GC-MS analysis. The injection was performed manually, with the GC-MS set to QI SIM mode for m / z 16 and m / z 15 and a split ratio of 1:50. The column was kept at 30 °C for 2 min. Peak separation of all headspace gases was not achieved but methane could be identified using ion m / z 15 and 16. Methane quantification was performed by integration of peak areas of ion m / z 15 and standard curve based on natural gas standards. To calculate the amount of methane reduction in in vitro rumen trials, peak areas of control samples without any additive were compared to samples with additive.

[0151] At each concentration of bromoform, the BSB samples gave rise to higher levels of methanogenesis inhibition (see Figure 4). This shows that there are methanogenesis inhibitors other than bromoform present as a result of treating biomass with the bromination solution.

[0152] In a second experiment demonstrating the presence of methanogenesis inhibitors other than bromoform, biomass (500 pL of liquid corn extract) was brominated using 62.5 mM hypobromite solution for 2 mins at room temperature with no agitation prior to the addition of 5 M sodium thiosulfate. The liquid corn extract was prepared by grinding commonly available corn kernels using mortar and pestle. The resulting mixture of solids and liquids was then centrifuged at 4000 rpm for 4 mins to remove insoluble solids. The supernatant was used as liquid extract in the experiment. Brominated biomass mixtures were heated at 60 °C for 1 hr to evaporate bromoform, but retain any less volatile active ingredients. After this evaporation process, bromoform was quantified using GC-MS (as described above), and an identical amount was added to the same mass of material that had not been treated with bromination solution or sodium thiosulfate. Each sample, with identical bromoform amounts, was then dosed into in vitro rumens to test for any inhibition of methanogenesis (as described above).

[0153] As can be seen from Figure 5, in each case, the samples that had been treated with brominating solution gave rise to higher levels of inhibition compared to bromoform-spiked biomass samples, indicating that other active ingredients were present in the brominated biomass. Althoughsodium thiosulfate-treated corn also inhibited methane production, the level of inhibition was less than that seen between bromination solution-treated and non-treated biomass.

[0154] In a third demonstration of biomass bromination to produce bromoform and nonbromoform brominated methanogenesis inhibitors, 0.25 g samples of biomass were treated with different concentrations of hypobromite solution (33.3 mM labelled “4”, 41.7 mM labelled “5”, 50 mM labelled “6”, and 83 mM labelled “10”) at room temperature and no agitation for 4 hours. The reaction was halted by addition of 5 M sodium thiosulfate. The concentration of bromoform from each treatment was normalised to the indicated levels, and tested for methanogenesis inhibition using a methane measurement from in vitro rumen cultures (as described above). As shown in Figure 6, different sources of biomass can be brominated to inhibit methanogenesis. Further, the dependence of this effect on non-bromoform molecules is demonstrated, since differential methanogenesis inhibition was seen between the different hypobromite solution concentrations used, even though bromoform was normalised to the same concentration within biomass types. This effect was particularly evident when yeast was brominated with 83 mM and 50 mM of hypobromite solution and bromoform levels normalised to 5 M.

[0155] Example 3

[0156] Inhibition of methanogenesis in ruminants with brominated biomass

[0157] 6000 g of oats was combined with 1500 g water and mixed for 30 minutes. To the resulting mixture was added 420 g NaOH slowly over the course of 1 hr, followed by slow addition of 1500 g DBDMH over the course of 1 hr. Both steps are exothermic. The mixture was stirred for 24 hrs. This produced 5.592 mg of bromoform per gram of total mixture (total of 50.32 g of bromoform for the entire reaction). 673 g of -cyclodextrin was then added and stirred for 12 hrs. 100 g of chemical bromoform was then added and mixed for an additional 24 hrs, bringing the final concentration to 15.54 mg bromoform per gram of mixture.

[0158] This mixture was fed to ruminants (cattle) at four inclusion rates:(1) Tl: 0 g per kg dry matter intake (DMI);(2) T2: 1.15 g per kg DMI;(3) T3: 2.30 g per kg DMI;(4) T4: 4.59 g per kg DMI.

[0159] A total of 12 ruminants were included in the trial, three per treatment condition. The animals were fed their allocated treatment for 27 days, with measurement continuing for an additional three days post treatment administration. Methane and carbon dioxide emissions were measured using four Agscent Air units (1 per treatment group), model GHG2100 (Carwoola, NSW, Australia). The results are shown in Figure 7.

[0160] The units were utilised in four of the 12 animal pens. The four corner pens were used to eliminate the potential of cross sample contamination, or an RFID read from a calf in a neighbouring pen. The AgScent units themselves measured 40 cm x 20 cm x 15 cm in size, and contained two optical sensors - one which was a laser diode to measure methane and the other a nondispersive infrared (NDIR) optical sensor to measure carbon dioxide. The units were mounted in the alley way, with a sample tube with an outer diameter of 6 mm positioned from the unit to inside the wheelie bin feeder at a height approximately 60 cm from the base of the wheelie bin.

[0161] The sample tube had a sampling flow rate of 1 L / min. An RFID panel reader was secured on the outside of the right-hand side of the wheelie bin feeder at the height of the animal’s ear to detect when a calf was eating from the feeder. When a calf was within 30 cm of the sampling tube for approximately 30 seconds or more, a methane peak, if present, was detected. A methane peak caused by a calf should also coincide with a peak in carbon dioxide.

[0162] Calves were rotated between pens on a two-day basis during the adaptation period and on a three-day basis during the measurement period to allow measurements of methane from each calf for a total of nine days (3 x 3-day periods). The AgScent sample tubes were continuously collecting samples of air that passed through the AgScent sensors therefore baseline measures of methane and carbon dioxide were able to be determined based on when calves were not near the feeder. Baseline measures were also collected during the adaptation and post-treatment measurement periods. Study and AgScent personnel received SMS alerts when there was no air flow into the units or data flow to the processing centre. Data were monitored and downloaded through a website dashboard.

[0163] The present disclosure is useful as it provides an inexpensive and scalable method of producing halogenated carbon compounds. Increased amounts of a variety of halogenated compounds may be produced from cheap organic materials, such as biomass. The produced halogenated carbon compounds may find use in animal feed and animal supplements for reducing methane emissions in animals, such as ruminant animals. The produced variety of halogenated compounds may be more effective than bromoform alone in reducing methane emissions in animals, such as ruminant animals.

[0164] Reference to any prior art in this specification is not, and should not be taken as, an acknowledgement or any form of suggestion that such prior art forms part of the common general knowledge.

[0165] In some cases, a single embodiment may, for succinctness and / or to assist in understanding the scope of the disclosure, combine multiple features. It is to be understood that in such a case, these multiple features may be provided separately (in separate embodiments), or in any other suitable combination. Alternatively, where separate features are described in separate embodiments, these separate features may be combined into a single embodiment unless otherwise stated or implied. This also applies to the claims which can be recombined in any combination. That is a claim may be amended to include a feature defined in any other claim. Further a phrase referring to "at least one of" a list of items refers to any combination of those items, including single members. As an example, "at least one of: a, b, or c" is intended to cover: a, b, c, a-b, a-c, b-c, and a-b-c.

[0166] It will be appreciated by those skilled in the art that the disclosure is not restricted in its use to the particular application or applications described. Neither is the present disclosure restricted in its preferred embodiment with regard to the particular elements and / or features described or depicted herein. It will be appreciated that the disclosure is not limited to the embodiment or embodiments disclosed, but is capable of numerous rearrangements, modifications and substitutions without departing from the scope as set forth and defined by the following claims.

Claims

CLAIMS1. A method of producing a plurality of halogenated carbon compounds, the method comprising contacting a source of electron-rich carbon compounds with a composition comprising hypohalite ions under conditions and for a time sufficient to produce the plurality of halogenated carbon compounds.

2. The method of claim 1, wherein the source of electron-rich carbon compounds is an organic material.

3. The method of claim 2, wherein the organic material is biomass.

4. The method of claim 3, wherein the biomass is a microorganism, a plant material, or a combination thereof.

5. The method of any one of the preceding claims, wherein the electron-rich carbon compounds comprise a ketone.

6. The method of claim 5, wherein the ketone is a methyl ketone.

7. The method of claim 6, wherein the methyl ketone has the structure:wherein R1is selected from the group consisting of H, halogen, NO2, CN, NH2, optionally substituted Ci-C alkyl, optionally substituted C2-Ci2alkenyl, optionally substituted C2-Ci2alkynyl, optionally substituted C2-Ci2heteroalkyl, optionally substituted Cs-C cycloalkyl, optionally substituted C2-Ci2heterocycloalkyl, optionally substituted C2-Ci2heterocycloalkenyl, optionally substituted C(,-C is aryl, optionally substituted Ci-Cisheteroaryl, optionally substituted Ci-C alkyloxy, optionally substituted C2-Ci2alkenyloxy, optionally substituted C2-Ci2alkynyloxy, optionally substituted C2-Ci2heteroalkyloxy, optionally substituted C3-Ci2cycloalkyloxy, optionally substituted C3-Ci2cycloalkenyloxy, optionally substituted Ci-Ci2heterocycloalkyloxy, optionally substituted C2-Ci2heterocycloalkenyloxy, optionally substituted Ce-Cisaryloxy, optionally substituted Ci-Cisheteroaryloxy, optionally substituted Ci- Ci2alkylamino, SR2, SO3H, SO2NH2, SO2R2, SONH2, SOR2, COR2, COOH, COOR2, CON R2R3, NR2COR3, NR2COOR3, NR2SO2R3, N R2CON R2R3, N R2R3, and acyl; wherein each R2andR3isindependently selected from the group consisting of: H, optionally substituted Ci-Ci2alkyl, optionally substituted CT-C alkcnyl, optionally substituted C2-Ci2alkynyl, optionally substituted C2-Ci2heteroalkyl, optionally substituted C3-Ci2cycloalkyl, optionally substituted C3-Ci2cycloalkenyl, optionally substituted C2-Ci2heterocycloalkyl, optionally substituted C2-Ci2heterocycloalkenyl, optionally substituted Cr-C- isaryl, and optionally substituted Ci-Cisheteroaryl.

8. The method of any one of the preceding claims, wherein the halogenated carbon compounds comprise haloform.

9. The method of claim 8, wherein the haloform is bromoform.

10. The method of any one of the preceding claims, wherein the amount of hypohalite ions in the composition is at least between about 0.001 mol and about 0.05 mol per gram of the source.

11. The method of any one of the preceding claims, wherein the composition is a solution.

12. The method of claim 11, wherein the solution is an aqueous solution.

13. The method of claim 12, wherein the hypohalite ions are hypobromite ions or hypochlorite ions, or a combination thereof.

14. The method of any one of the preceding claims, the method further comprising extracting one or more of the halogenated carbon compounds after producing the plurality of halogenated carbon compounds.

15. An animal supplement comprising the plurality of halogenated carbon compounds produced by the method of any one of the preceding claims, or an extract thereof.

16. An animal feed comprising the supplement or extract of claim 15.

17. A method of reducing methane emissions of an animal, comprising administering an effective amount of the supplement of claim 15 or the animal feed of claim 16 to the animal.

18. The method of claim 17, wherein the animal is a ruminant animal.

19. The method of claim 17 or claim 18, wherein the effective amount is about 0.01% to about 5% dry weight of the animal's diet.

20. A composition for producing a plurality of halogenated carbon compounds, the composition comprising: a source of electron-rich carbon compounds; and a composition comprising hypohalite ions.

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