Methanogenesis inhibitor formulations

Abstract

The present technology generally relates to a feed supplement pre-mix comprising: between about 50% and about 90% dewatered Asparagopsis spp.; and between about 15% and about 50% edible oil, wherein the dewatered Asparagopsis spp. has a moisture content of between about 50% and about 80%.

Description

METHANOGENESIS INHIBITOR FORMULATIONSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. provisional patent application No. 63 / 659,565, filed on June 13, 2024; the content of all of which is herein incorporated in entirety by reference.TECHNICAL FIELD

[0002] The present technology generally relates to formulations of biomass comprising bromoform for the inhibition of methanogenesis in ruminants.BACKGROUND

[0003] Methanogenesis (i.e., the production of methane by ruminant animals) is a major contributor to global greenhouse gas emissions. Scientific literature has demonstrated reductions in methane (CH4) production of beef cattle and dairy cows when halogenated compounds are fed as part of the diet (see for example Roque 2020, Red seaweed (Asparagopsis taxiformis) supplementation reduces enteric methane by over 80 percent in beef steers, doi.org / 10.1101 / 2020.07.15.204958 “Some haloalkanes are structural analogs of CH4 and therefore competitively inhibit the methyl transfer reactions that are necessary in CH4 biosynthesis. The CH4 analogues include bromochloromethane (BCM), bromoform and chloroform and have been proven to be the most effective feed additives for reducing CH4 production”, incorporated herein by reference). There are two potential sources for halogenated compounds: artificial synthesis and natural production, (Machado 2018, doi.org / 10.1007 / s00248-017-1086-8, incorporated herein by reference). Supplementation with gametophytes of Asparagopsis taxiformis comprising 7.8 mg / g of bromoform by dry weight has been found to mitigate an 80% reduction of methane gas production over a 147-day period in steers fed a 0.5% seaweed supplemented low forage total mixed ration (LF-TMR).

[0004] Despite its efficacy, the potential of Asparagopsis taxiformis (AT) as a feed additive to inhibit methanogenesis in ruminant animals is constrained by several factors. These include its unpleasant odor, high iodine content, epiphytic nature, and the lack of capacity,especially in male AT specimens, to synthesize material concentrations of the halogenated compounds.

[0005] CSIRO, Meat and Livestock Australia, and James Cook University are also the holders of US Patent No: 9,980,995, incorporated herein by reference. This patent describes the administration of an effective amount of red marine macroalgae as a feed supplement, with the result being a reduction in methane production. The red macroalgae form of the species, Asparagopsis taxiformis, is specifically identified as one lifestage morphotype of the species effective in this method. This patent also mentions the filamentous tetrasporophyte lifestage of Asparagopsis taxiformis.

[0006] US Patent Application No: 2019 / 0174793, incorporated herein by reference, discloses a method for improving the growth performance of a livestock animal in various farming systems, including providing a red marine macroalgae to the farming systems to enable consumption of the red marine macroalgae.

[0007] US Patent Application No: 2019 / 0174793, incorporated herein by reference, describes a formulation such that the animal is provided with of red marine macroalgae per day for animals maintained at pasture and states that amounts to between about 1-5% of algae on a dry matter basis or 1-3% on an organic matter basis per day. US Patent Application No: 2019 / 0174793, incorporated herein by reference, also discloses providing an animal with between about 200-600 g / day of algae for animals on a finishing diet in feedlots.

[0008] Stabilization of Asparagopsis with oil is disclosed in US Patent Application No:2022 / 0031780, incorporated herein by reference.

[0009] Asparagopsis and oil combinations are also disclosed in PCT Patent ApplicationsWO 2023279152, WO 2024 / 073579 and WO 2022 / 232876, both incorporated herein by reference.

[0010] Generation of bromoform producing yeast organisms is disclosed in PCT PatentApplication Publication WO 2020 / 243792, incorporated herein by reference.

[0011] The use of exogenous organohalide enriched macroalgal substrates are disclosed in PCT Patent Application Publication WO 2023 / 015042, incorporated herein by reference.

[0012] Non-algal or yeast derived halocarbon compound compositions for reducing methanogenesis are disclosed in PCT Patent Application Publication WO 2023 / 010170, incorporated herein by reference.

[0013] United States Patent US12,116,608 discloses the use of haloperoxidases to make antimethanogenic halide compounds, incorporated herein by reference.SUMMARY OF DISCLOSURE

[0014] In one embodiment, the present technology provides for a feed supplement premix comprising: between about 50% and about 90% dewatered Asparagopsis spp. and between about 15% and about 50% edible oil, wherein the dewatered Asparagopsis spp. has a moisture content of between about 50% and between about 80%.

[0015] In one embodiment, the present technology provides for a feed supplement premix comprising: between about 50% and about 90% dewatered Asparagopsis spp. and between about 15% and about 50% edible oil, wherein the dewatered Asparagopsis spp. has a moisture content of between about 60% and about 80%.

[0016] In one embodiment, the present technology provides for a feed supplement premix comprising: between about 50% and about 90% dewatered Asparagopsis spp. and between about 15% and about 50% edible oil, wherein the dewatered Asparagopsis spp. has a moisture content of between about 65% and about 75%.

[0017] In one embodiment, the present technology provides for a feed supplement premix comprising: between about 60% and about 80% dewatered Asparagopsis spp. and between about 15% and about 50% edible oil, wherein the dewatered Asparagopsis spp. has a moisture content of between about 65% and about 75%.

[0018] In one embodiment, the present technology provides for a feed supplement premix comprising: between about 65% and about 75% dewatered Asparagopsis spp. and between about 15% and about 50% edible oil, wherein the dewatered Asparagopsis spp. has a moisture content of between about 65% and about 75%.

[0019] In one embodiment, the present technology provides for a feed supplement premix comprising: between about 50% and about 90% dewatered Asparagopsis spp., between about 15% and about 50% edible oil, between about 0.1% and about 5% xanthan gum, wherein the dewatered Asparagopsis spp. has a moisture content of between about 50% and about 80%.

[0020] In one embodiment, the present technology provides for a feed supplement premix comprising: between about 50% and about 90% dewatered Asparagopsis spp., between about 15% and about 50% edible oil, between about 0.1% and about 5% xanthan gum, wherein the dewatered Asparagopsis spp. has a moisture content of between about 60% and about 80%.

[0021] In one embodiment, the present technology provides for a feed supplement premix comprising: between about 50% and about 90% dewatered Asparagopsis spp., between about 15% and about 50% edible oil, and between about 0.1% and about 5% xanthan gum, wherein the dewatered Asparagopsis spp. has a moisture content of between about 65% and about 75%.

[0022] In one embodiment, the present technology provides for a feed supplement premix comprising: between about 60% and about 80% dewatered Asparagopsis spp., between about 15% and about 50% edible oil, and between about 0.1% and about 5% xanthan gum, wherein the dewatered Asparagopsis spp. has a moisture content of between about 65% and about 75%.

[0023] In one embodiment, the present technology provides for a feed supplement premix comprising: between about 65% and about 75% dewatered Asparagopsis spp., between about 15% to 50% edible oil, and between about 0.1% and about 5% xanthan gum, wherein the dewatered Asparagopsis spp. has a moisture content of between about 65% and about 75%.

[0024] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 50% and about 90% of a feed supplement pre-mix, between about 5% and about 50% carrier, between about 0.5% and about 10% binder, and between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 10% and about 50% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0025] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 60% and about 85% of a feed supplement pre-mix, between about 5% and about 50% carrier, between about 0.5% and about 10% binder, and between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 10% and about 50% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0026] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 70% and about 80% of a feed supplement pre-mix, between about 5% and about 50% carrier, between about 0.5% and about 10% binder, and between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 10% and about 50% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0027] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 50% and about 90% of a feed supplement pre-mix, between about 5% and about 50% carrier, between about 0.5% and about to 10% binder, and between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 10% and about 50% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0028] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 60% and about 85% of a feed supplement pre-mix, between about 5% and about 50% carrier, between about 0.5% and about 10% binder, and between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 10% and about 50% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0029] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 60% and about 85% of a feed supplement pre-mix, between about 5%and about 50% carrier, between about 0.5% and about 10% binder, and between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 10% and about 50% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0030] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 70% and about 80% of a feed supplement pre-mix, between about 5% and about 50% carrier, between about 0.5% and about 10% binder, and between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 10% and about 50% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0031] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 70% and about 80% of a feed supplement pre-mix, between about 5% and about 50% carrier, between about 0.5% and about 10% binder, and between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 10% and about 50% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0032] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 70% and about 80% of a feed supplement pre-mix, between about 5% to 50% carrier, between about 0.5% and about 10% binder, and between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis spp. biomass and 20% to 40% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0033] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 70% and about 80% of a feed supplement pre-mix, between about 5% and about 50% carrier, between about 0.5% and about 10% binder, and between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about50% and about 90% dewatered Asparagopsis spp. biomass and between about 25% and about 35% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0034] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 60% and about 85% of a feed supplement pre-mix, between about 5% and about 50% carrier, between about 0.5% and about 10% binder, and between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 20% and about 40% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0035] In one embodiment, the present technology provides for a feed supplement mix comprising: between about 60% and about 85% of a feed supplement pre-mix, between about 5% and about 50% carrier, between about 0.5% and about 10% binder, and 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 25% and about 35% edible oil; and wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0036] In one embodiment, the present technology provides for a feed supplement shot comprising: between about 40% and about 60% Asparagopsis spp. biomass, between about 17.5% and about 27.5% edible oil, between about 20% and about 30% carrier, between about 0.5% and about 3% xanthan gum, and between about 0.1% and about 2% palatability enhancer, wherein the feed supplement shot is formed by extrusion or pressing, wherein the Asparagopsis spp. biomass has a water content of between about 70% and about 80%, and wherein the feed supplement shot has a moisture content of between about 30% and about 45%.

[0037] In one embodiment, the present technology provides for a feed supplement shot comprising: between about 65% and about 75% Asparagopsis spp. biomass, between about 20% and about 30% edible oil, between about 10% and about 20% carrier, between about 0.5% and about 3% xanthan gum, and between about 0.1% and about 2% palatability enhancer, wherein the feed supplement shot is formed by extrusion or pressing, wherein the Asparagopsis spp. biomasshas a water content of between about 70% and about 80%, and wherein the feed supplement shot has a moisture content of between about 40% and about 50%.BRIEF DESCRIPTION OF DRAWINGS

[0038] Figure 1 is a graph illustrating bromoform concentration over time in feed suppl ement-pre-mix with 20%, 35%, and 50% final oil inclusion rates from Example 1. Feed supplement-pre-mix samples were stored in closed glass vials at room temperature, N=3.

[0039] Figure 2 is a graph illustrating bromoform retention (in comparison to bromoform concentration calculated based on bromoform level in source biomass) in feed supplement-pre-mix with 20%, 35%, and 50% final oil inclusion rates (light gray circles, gray triangles, and black squares respectively) From Example 1. Samples were stored in closed glass vials at room temperature. N=3.

[0040] Figure 3 is a graph illustrating Bromoform retention in the feed supplement premix stored in farm conditions over 22 weeks from Example 2. Three similar storage buckets were sampled routinely to monitor feed supplement pre-mix shelflife. Six samples were taken from each bucket at each time point. Error bars show the standard error of replicates (N=6).

[0041] Figure 4 is a graph illustrating bromoform retention through pelletization forAsparagopsis feed supplement shot pellets based on 75% and 85% feed supplement pre-mix inclusion (formula 1 and formula 2 in Table 1 respectively) from Example 3. Gray diamonds and black circles represent recipe 1 and 2 respectively. n=6.

[0042] Figure 5 is a graph illustrating bromoform retention through storage forAsparagopsis feed supplement shot pellets based on 75% (gray diamonds) and 85% (gray squares) feed supplement pre-mix inclusion (formula 1 and formula 2 in Table 1 respectively), compared to bromoform retention in freeze dried Asparagopsis powder (black circles) from Example 3. n=9

[0043] Figure 6 is a graph illustrating Bromoform retention for Brominata pellets stored in farm storage over 8 weeks from Example 4. Three similar pellet bags were sampled routinely to monitor pellet shelflife. Six samples were taken from each bag at each time point. Error bars show the standard error of replicates (N=6).

[0044] Figure 7 are photographs of the results of the shelf-life stability test of theAsparagopsis based feed supplement shot pellets versus kelp control from Example 6.

[0045] Figure 8 is a graph showing the results of the bromoform enhancement experiment outlined in Example 9 and shown in tabular form in Table 16.

[0046] Figure 9 is a graph showing the results of the bromoform enhancement experiment outlined in Example 10 and shown in tabular form in Table 17.DETAILED DESCRIPTION OF TECHNOLOGY

[0047] The present technology is explained in greater detail below. This description is not intended to be a detailed catalog of all the different ways in which the technology may be implemented, or all the features that may be added to the instant technology. For example, features illustrated with respect to one embodiment may be incorporated into other embodiments, and features illustrated with respect to a particular embodiment may be deleted from that embodiment. In addition, numerous variations and additions to the various embodiments suggested herein will be apparent to those skilled in the art in light of the instant disclosure which variations and additions do not depart from the present technology. Hence, the following description is intended to illustrate some particular embodiments of the technology, and not to exhaustively specify all permutations, combinations and variations thereof.

[0048] As used herein, the singular form “a”, “an” and “the” include plural referents unless the context clearly dictates otherwise.

[0049] The recitation herein of numerical ranges by endpoints is intended to include all numbers subsumed within that range (e.g., a recitation of 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.80, 4, 4.32, and 5).

[0050] The term “about” is used herein explicitly or not, every quantity given herein is meant to refer to the actual given value, and it is also meant to refer to the approximation to such given value that would reasonably be inferred based on the ordinary skill in the art, including equivalents and approximations due to the experimental and / or measurement conditions for such given value. For example, the term “about” in the context of a given value or range refers to a value or range that is within 20%, preferably within 15%, more preferably within 10%, more preferablywithin 9%, more preferably within 8%, more preferably within 7%, more preferably within 6%, and more preferably within 5% of the given value or range.

[0051] The expression “and / or” where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. For example, “A and / or B” is to be taken as specific disclosure of each of (i) A, (ii) B and (iii) A and B, just as if each is set out individually herein.

[0052] When a quantifier such as “less than”, “more than”, or “greater than” modifies a comma delimited list of numbers or quantities, or ratios, such as “1, 2.5, 3, ...” or “one, two, three. .. ” the intention is that the quantifier applies to every member of the list. For example, “alpha is greater than about 1, 2, or 3, or 4” means “alpha is greater than about 1, or, alpha is greater than about 2 or alpha is greater than about 3.”

[0053] As used herein, the term “feed supplement pre-mix” refers to a composition comprising, biomass comprising bromoform, edible oil, and optionally, a binder. This biomass may in some instances be de-watered Asparagopsis spp. biomass. In other instances, it may be bromoform producing yeast biomass. In still other instances, it may be a biomass which has been treated with biologically or synthetically produced bromoform.

[0054] As used herein, the term “feed supplement mix” refers to a composition comprising feed supplement pre-mix, carrier, a binder, and optionally, a palatability enhancer.

[0055] As used herein, the term “shot” refers to a feed supplement form comprising feed supplement mix that has been mechanically or thermally processed to a solid or semisolid particulate flowable consistency. Non limiting examples of forms of shot include pellets, nuggets, crumbles, flakes, powders, granules, and kibble.

[0056] As used herein, the term “enhanced feed supplement pre-mix” refers to a composition comprising biomass further comprising bromoform, edible oil, and optionally, a binder that has been treated with a carboxylic acid and a peroxide source. This biomass may in some instances be de-watered Asparagopsis spp. biomass. In other instances, it may be bromoform producing yeast biomass. In other instances, it may be bromoform producing organism biomass.

[0057] As used herein, the term Asparagopsis spp. refers to any species of Asparagopsis algae such as Asparagopsis taxiformis and Asparagopsis armata. The terms Asparagopsis spp. and Asparagopsis may be used interchangeably.

[0058] As used herein, the term “enhanced feed supplement mix” refers to a composition comprising enhanced feed supplement pre-mix, carrier, a binder, and optionally, a palatability enhancer.

[0059] As used herein, the term “enhanced shot” refers to a feed supplement form comprising enhanced feed supplement mix that has been mechanically or thermally processed to a solid or semisolid particulate flowable consistency. Non limiting examples of forms of enhanced shot include pellets, nuggets, crumbles, flakes, powders, granules, and kibble.

[0060] As used herein, the term “enriched feed supplement pre-mix” refers to a composition comprising, biomass, edible oil, and optionally, a binder that has been treated with synthetic or naturally derived bromoform. This biomass may in some instances be de-watered Asparagopsis spp. biomass. In other instances, it may be bromoform producing yeast biomass. In other instances, it may be bromoform producing organism biomass.

[0061] As used herein, the term “enriched feed supplement mix” refers to a composition comprising enriched feed supplement pre-mix, carrier, a binder, and optionally, a palatability enhancer.

[0062] As used herein, the term “enriched shot” refers to a feed supplement form comprising enriched feed supplement mix that has been mechanically or thermally processed to a solid or semisolid particulate flowable consistency. Non limiting examples of forms of enhanced shot include pellets, nuggets, crumbles, flakes, powders, granules, and kibble.

[0063] As used herein, the term “carboxylic acid” refers to: an organic acid comprising at least one carboxylate moiety, or an ester or salt thereof. Non-limiting examples of carboxylic acids include formic acid, acetic acid, propionic acid, butyric acid, isobutyric acid, pentanoic acid, 2- methylbutanoic acid, 3 -methylbutanoic acid, oxalic acid, malonic acid, succinic, lactic, pyruvic and the like. Non-limiting examples of salt forms of carboxylic acids include lithium, sodium, calcium, magnesium, potassium and ammonium. Non-limiting ester forms of carboxylic acids includemethyl, ethyl, propyl, iso-propyl, butyl, iso-butyl, sec-butyl, tert-butyl, and the like. In some embodiments, the carboxylic acid is an alpha-keto carboxylic acid. In other embodiments, the carboxylic acid is a beta-keto carboxylic acid.

[0064] As used herein, the term “peroxide source” refers to alkaline metal, alkaline earth metal, aluminum, zinc, and ammonium salts of perborate, peroxide, percarbonate and peroxy monosulfate. Non limiting examples of peroxide sources include benzoyl peroxide, sodium peroxide, calcium peroxide, magnesium peroxide potassium peroxide, peracetic acid, carbamide peroxide, potassium peroxymonosulfate, and ozone. A peroxide source may also mean urea hydrogen peroxide or hydrogen peroxide.

[0065] As used herein, the term “bromide source” means an alkaline metal or alkaline earth metal salt of bromide, ammonium bromide, or magnesium bromide.

[0066] As used herein, the expression “typical farm conditions” encompass a condition at which the average temperature is of about 15 deg. C. with a minimum of 10 deg. C and a maximum of 22 deg. C. or preferably a condition at which the average temperature is of 13.6 deg. C. with a minimum of 9 deg. And a maximum of 21.1 deg. C.

[0067] The present technology also includes the corresponding enriched and enhanced versions of feed supplement pre-mix, feed supplement mix and feed supplement shot that are disclosed in the embodiments herein.

[0068] Asparagopsis biomass that has been cultivated under conditions to maximize its bromoform content and bromoform to iodine content ratio is preferred, such as the Asparagopsis disclosed in W02021 / 205420A1 which is incorporated herein in its entirety by reference.

[0069] The key advantage of the present technology is the provision of highly shelf stable ruminant animal feed supplements that inhibit methanogenesis in an economically advantageous manner.

[0070] The main issue with the production of Asparagopsis based supplements is the need for thorough drying of the algae to prevent spoilage. The prevention of bromoform loss through drying specifically requires freeze drying which is highly energy intensive. The present technology addresses this issue by providing formulations that remain chemically and biologically stable forweeks in typical farm storage conditions, but that do not need any drying beyond quick and inexpensive dewatering.Feed supplement pre-mix

[0071] In one embodiment, the present technology provides for a feed supplement premix composition comprising dewatered Asparagopsis spp., biomass, edible oil and optionally, xanthan gum.

[0072] In another embodiment, the Asparagopsis spp. biomass is Asparagopsis taxiformis biomass o Asparagopsis armata biomass.

[0073] In a preferred embodiment, the Asparagopsis spp. biomass is a tetrasporophyte lifestage of Asparagopsis spp.

[0074] Dewatering may be accomplished by any method known in the art such as centrifugation, spin drying, drum dryers, belt dryers, screw press (such as CP6 screw press, Vincent Corporation, USA), oil injection screw press, gravity drainage, open air drying, and solar assisted open air drying.

[0075] Typically, using a CP6 screw press, one would apply 80 PSI to the discharge cone to achieve 75% residual moisture content, or, more generally, 20-100 PSI to achieve between 75% and 85% residual moisture content.

[0076] In one embodiment, the present technology provides for a feed supplement premix composition comprising bromoform producing yeast biomass, edible oil and optionally, xanthan gum.

[0077] In one embodiment, the present technology provides for a feed supplement premix composition comprising biomass which has been treated with biologically or synthetically produced bromoform, edible oil and optionally, xanthan gum.

[0078] In one embodiment, the feed supplement pre-mix has a bromoform to iodine ratio of at least 2 to 1 wt / wt.

[0079] In a preferred embodiment, the feed supplement pre-mix has a bromoform to iodine ratio of at least 5 to 1 wt / wt.

[0080] In an even more preferred embodiment, the feed supplement pre-mix has a bromoform to iodine ratio of at least 9 to 1 wt / wt.

[0081] In one embodiment, the present technology provides for a feed supplement premix composition comprising between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 50% and about 10% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 50% to 80% residual moisture.

[0082] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 50% and about 10% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0083] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 50% and about 90% dewatered Asparagopsis spp. biomass and between about 50% and about 10% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0084] In one embodiment, the present technology provides for a feed supplement premix composition comprising between about 60% and about 80% dewatered Asparagopsis spp. biomass and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0085] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 60% and about 80% dewatered Asparagopsis spp. biomass and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0086] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 60% and about 80% dewatered Asparagopsis spp. biomass and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0087] In one embodiment, the present technology provides for a feed supplement premix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0088] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0089] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0090] In one embodiment, the present technology provides for a feed supplement premix composition comprising between about 50% and about 90% dewatered Asparagopsis spp. biomass, between about 0.01% and about 10% xanthan gum, and between about 50% and about 10% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0091] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 50% and about 90% dewatered Asparagopsis biomass, between about 0.01% and about 10% xanthan gum, and between about 50% and about 10% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0092] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 50% and about 90% dewatered Asparagopsis spp. biomass, between about 0.01% and about 10% xanthan gum, and between about 50% and about 10% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0093] In one embodiment, the present technology provides for a feed supplement premix composition comprising between about 60% and about 80% dewatered Asparagopsis spp. biomass, between about 0.01% and about 10% xanthan gum, and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises 50% and about 80% residual moisture.

[0094] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 60% and about 80% dewatered Asparagopsis spp. biomass, between about 0.01% and about 10% xanthan gum, and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0095] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 60% and about 80% dewatered Asparagopsis spp. biomass, between about 0.01% and about 10% xanthan gum, and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0096] In one embodiment, the present technology provides for a feed supplement premix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass, between about 0.01% and about 10% xanthan gum, and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0097] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass, between about 0.01% and about 10% xanthan gum, and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0098] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass, between about 0.01% and about 10% xanthan gum, and between about35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0099] In one embodiment, the present technology provides for a feed supplement premix composition comprising between about 50% and about 90% dewatered Asparagopsis spp. biomass, between about 0.5% and about 6% xanthan gum, and between about 50% and about 10% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0100] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 50% and about 90% dewatered Asparagopsis spp. biomass, between about 0.5% and about 6% xanthan gum, and between about 50% and about 10% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0101] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 50% and about 90% dewatered Asparagopsis spp. biomass, between about 0.5% and about 6% xanthan gum, and between about 50% and about 10% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0102] In one embodiment, the present technology provides for a feed supplement premix composition comprising between about 60% and about 80% dewatered Asparagopsis spp. biomass, between about 0.5% and about 6% xanthan gum, and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0103] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 60% and about 80% dewatered Asparagopsis spp. biomass, between about 0.5% and about 6% xanthan gum, and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0104] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 60% and about 80% dewatered Asparagopsis spp. biomass, between about 0.5% and about 6% xanthan gum, and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0105] In one embodiment, the present technology provides for a feed supplement premix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass, between about 0.5% and about 6% xanthan gum, and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0106] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass, between about 0.5% and about 6% xanthan gum, and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0107] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass, between about 0.5% and about 6% xanthan gum, and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0108] In one embodiment, the present technology provides for a feed supplement premix composition comprising between about 50% and about 90% dewatered Asparagopsis spp. biomass, between about 1% and about 5% xanthan gum, and between about 50% and about 10% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0109] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 50% and about 90% dewatered Asparagopsis spp. biomass, between about 1% and about 5% xanthan gum, and between about 50% and about 10%edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0110] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 50% and about 90% dewatered Asparagopsis spp. biomass, between about 1% and about 5% xanthan gum, and between about 50% and about 10% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0111] In one embodiment, the present technology provides for a feed supplement premix composition comprising 60% and about 80% dewatered Asparagopsis spp. biomass, between about 1% and about 5% xanthan gum, and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0112] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 60% and about 80% dewatered Asparagopsis spp. biomass, between about 1% and about 5% xanthan gum, and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0113] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 60% and about 80% dewatered Asparagopsis spp. biomass, between about 1% and about 5% xanthan gum, and between about 40% and about 20% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.[001 14] In one embodiment, the present technology provides for a feed supplement premix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass, between about 1% and about 5% xanthan gum, and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 50% and about 80% residual moisture.

[0115] In a preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising 65% and about 75% dewatered Asparagopsis spp. biomass, between about 1% and about 5% xanthan gum, and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

[0116] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass, between about 0.5% and about 6% xanthan gum, and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

[0117] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises at least 65% residual moisture with the feed supplement premix composition further comprising between about 1% and about 5% xanthan gum.

[0118] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises at least 70% residual moisture with the feed supplement premix composition further comprising between about 1% and about 5% xanthan gum.

[0119] In a more preferred embodiment, the present technology provides for a feed supplement pre-mix composition comprising between about 65% and about 75% dewatered Asparagopsis spp. biomass and between about 35% and about 25% edible oil wherein the dewatered Asparagopsis spp. biomass comprises at least 75% residual moisture with the feed supplement premix composition further comprising between about 1% and about 5% xanthan gum.

[0120] The xanthan gum binder may be added as a powder to the dewatered biomass. In other cases, the xanthan gum binder may be added to the edible oil before mixing the dewatered biomass with the oil, or the xanthan gum binder may be added to the dewatered biomass-oil mixture.

[0121] In some embodiments, the binder may be an agent selected from the group consisting of Advanta-Gel™ P75, Batter Bind® S, Crisp Coat UC, Crisp Film®, Crystal Gum, Crystal TeX™ 627, Crystal Tex™ 644, Crystal Tex™ 648, Elastigel™ 1000J, Encapsul 855, Flojel® 60, Flojel® 65, Flojel® G, Hi-Set® 322, Hi-Set® 377, Hi-Set® C, Hi-Set® CHG, Hylon® V, Hylon® VII, Impression™, K4484, Melojel®, Nadex™ 772, National 0280, National 814, N- TACK®, Purity® 21D, Purity® TF, Superset® LV, Ultra-Set® LT, Dry-Tack® 250, Versa- Sheen™, Baka-Plus™, Baka-Snak®, Capsul®, Capsul® TA, Gel N Melt®, H-50, Hi-Cap™ 100, Hi-Cap™ 200, IF 131, Instant ClearGel®, Instant Pure-Flo, Instant Pure-Flo F, Instant Textaid-A, Instant Textra, National 104, National 1215, National 46, National 1517, National 5730, National 711, National 78-1551, N-Creamer 46, N-Flate, N-Lite™ LP, N-LOK®, N-LOK® 1930, Novation® 4600, Novation® 5600, Novation® 9460, Purity Gum 1773, Purity Gum 2000, Purity Gum 539, Purity Gum BE, Purity® HO, Stir-N-Set® FG, Text-Aid-A®, Textra® Plus, Ultra-Crisp CS, Ultra-Sperse® 2000, Ultra-Sperse® 5, Ultra-Sperse® A, Ultra- Sperse® M, Ultra-Tex 1, UltraTex 2, Ultra-Tex 2000, Ultra-Tex 3, Ultra-Tex 4, AbsorboHP, Amioca, Can-Fil®, Dry-Flo, Hoosier 5, National 150, National 1545, National 6912, National 77-1744, National 912, N-Zorbit® M, Purity® 21, Purity® 5, Purity® 825, Purity® 826, Purity® FC, Target brand tapioca, NU Mould™, Purity® 5S, Clearjel®, Clearjel® S, Colflo® 67, Firm-Tex®, Frigex® w, Hi FLO®, National 1333, National 1457, National 1658, National 4012, National 465, National 740, National Frigex, National Frigex HV, National® 320, Novation® 1600, Novation® 1900, Novation® 2300, Novation® 2600, Novation® 2700, Novation® 3300, Novation® 3600, Novation® 9230, Novation® 9260, Novation® 9270, Novation® 9330, Novation® 9360, Pure-Flo®, Purity® 270, Purity® 4, Purity® 420, Purity® 550, Purity® 660, Purity® 69, Purity® 87, Purity® Cloud, Purity® CSC, Purity® D, Purity® HPC, Purity® W, Thermflo®, Thermtex®, WNA. In still other embodiments, the bromoform bearing core may further comprise: Advanta-Gel™ P75, Batter Bind® S, Crisp Coat UC, Crisp Film®, Crystal Gum, Crystal TeX™ 627, Crystal Tex™ 644, Crystal Tex™ 648, Elastigel™ 1000J, Encapsul 855, Flojel® 60, Flojel® 65, Flojel® G, Hi-Set® 322, Hi-Set® 377, Hi-Set® C, Hi-Set® CHG, Hylon® V, Hylon® VII, Impression™, K4484, Melojel®, Nadex™ 772, National 0280, National 814, N-TACK®, Purity® 21D, Purity® TF, Superset® LV, Ultra-Set® LT, Dry-Tack® 250, Versa-Sheen™, Baka-Plus™, Baka-Snak®, Capsul®, Capsul® TA, Gel N Melt®, H-50, Hi-Cap™ 100, Hi-Cap™ 200, IF 131, Instant ClearGel®, Instant Pure-Flo, Instant Pure-Flo F, Instant Textaid-A, Instant Textra, National 104, National 1215, National 46, National 1517, National 5730, National 711, National 78-1551, N-Creamer 46, N-Flate, N-Lite™ LP, N-LOK®, N-LOK® 1930, Novation® 4600, Novation® 5600, Novation® 9460, Purity Gum 1773, Purity Gum 2000, Purity Gum 539, Purity Gum BE, Purity® HO, Stir-N-Set® FG, Text-Aid-A®, Textra® Plus, Ultra-Crisp CS, Ultra-Sperse® 2000, Ultra- Sperse® 5, Ultra-Sperse® A, Ultra-Sperse® M, Ultra-Tex 1, Ultra-Tex 2, Ultra-Tex 2000, UltraTex 3, Ultra-Tex 4, AbsorboHP, Amioca, Can-Fil®, Dry-Flo, Hoosier 5, National 150, National 1545, National 6912, National 77-1744, National 912, N-Zorbit® M, Purity® 21, Purity® 5, Purity® 825, Purity® 826, Purity® FC, Target brand tapioca, NU Mould™, Purity® 5S, Clearjel®, Clearjel® S, Colflo® 67, Firm-Tex®, Frigex® w, Hi FLO®, National 1333, National 1457, National 1658, National 4012, National 465, National 740, National Frigex, National Frigex HV, National® 320, Novation® 1600, Novation® 1900, Novation® 2300, Novation® 2600, Novation® 2700, Novation® 3300, Novation® 3600, Novation® 9230, Novation® 9260, Novation® 9270, Novation® 9330, Novation® 9360, Pure-Flo®, Purity® 270, Purity® 4, Purity® 420, Purity® 550, Purity® 660, Purity® 69, Purity® 87, Purity® Cloud, Purity® CSC, Purity® D, Purity® HPC, Purity® W, Thermflo®, Thermtex®, xanthan gum, alginate, a modified polysaccharide, guar gum, acacia gum and combinations thereof.

[0122] Non-limiting examples of edible oils used for making the feed supplement premix include: olive oil, canola oil, sunflower oil, soybean oil, corn oil, coconut oil, palm oil, peanut oil, sesame oil, safflower oil, avocado oil, grapeseed oil, rice bran oil, walnut oil, almond oil, flaxseed oil, pumpkin seed oil, mustard oil, cottonseed oil, macadamia nut oil, hemp seed oil, hazelnut oil, argan oil, pistachio oil, and black seed oil and food-grade mineral oil and combinations thereof.

[0123] In some embodiments, the edible oil may be a modified edible oil, such as, but not limited to, hydrogenated vegetable oil, partially hydrogenated vegetable oil, or brominated vegetable oil.Enhanced Feed supplement pre-mix

[0124] In some embodiments, the present technology provides for an enhanced feed supplement pre-mix.

[0125] In one embodiment, the present technology provides for a method of enhancing the bromoform content of a feed supplement pre-mix comprising:a) providing a feed supplement pre-mix; b) treating the feed supplement pre-mix with a peroxide source and optionally, a bromide source and / or a carboxylic acid; and c) obtaining enhanced feed supplement pre-mix.

[0126] In a preferred embodiment, the enhanced feed supplement pre-mix comprises about 50% water.

[0127] In an embodiment, the enhanced feed supplement pre-mix comprises between about 30-40%, 40-55%, 55-65%, 65-75%, or 75-85% water.

[0128] In another embodiment, the enhanced feed supplement pre-mix has a bromoform content of more than about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, or about 70% higher than the feed supplement pre-mix.

[0129] In a preferred embodiment, the treated biomass has a bromoform content of more than about 50% higher than the feed supplement pre-mix.

[0130] In a preferred embodiment, the peroxide source is sodium percarbonate.

[0131] In a preferred embodiment, the bromide source is sodium bromide.

[0132] In a preferred embodiment, the carboxylic acid is sodium acetate.

[0133] In a preferred embodiment, the biomass is treated only with a peroxide source.

[0134] In an embodiment, the peroxide source is not hydrogen peroxide.

[0135] In an embodiment, the carboxylic acid is not formic acid or a salt or ester thereof.Enriched Feed supplement pre-mix

[0136] In one embodiment, the present technology provides for a method of enriching the bromoform content of a feed supplement pre-mix comprising: a) providing a feed supplement pre-mix,b) treating the feed supplement pre-mix with synthetic or naturally produced bromoform, and c) obtaining enriched feed supplement pre-mix.Feed supplement mix

[0137] In one embodiment, the feed supplement mix comprises the feed supplement premix and a carrier.

[0138] In another embodiment, the feed supplement mix comprises the feed supplement pre-mix and a palatability enhancer.

[0139] In still another embodiment, the feed supplement mix comprises the feed supplement pre-mix, a carrier, and a palatability enhancer.

[0140] In one embodiment, the present technology provides for a feed supplement mix comprising a feed supplement pre-mix composition as described herein and a carrier product. The carrier product is typically a grain or legume-based product that is high in fiber and / or protein.

[0141] The carrier product is selected from the group consisting of wheat middlings, corn meal, soy hulls, and dried distiller’s grains.

[0142] In one embodiment, the feed supplement mix comprises a binder. Typically, the binder is a vegetable gum or a modified carbohydrate. Non-limiting examples include guar gum, alginate, and xanthan gum. The binder may be the binder that has been added to the feed supplement pre-mix as described herein, or the binder may be added to the feed supplement premix during the feed supplement mix production process. In some cases, the binder may be added during the feed supplement pre-mix production process as well as the feed supplement mix production process.

[0143] In some embodiments, two or more binders may be used in the feed supplement pre-mix, feed supplement mix, or feed supplement shot production processes.

[0144] In a preferred embodiment, the feed supplement mix comprises between about 0.5% and about 10% xanthan gum.

[0145] In a more preferred embodiment, the feed supplement mix comprises between about 1% and about 5% xanthan gum.

[0146] In an even more preferred embodiment, the feed supplement mix comprises between about 1% and about 3% xanthan gum.

[0147] In some embodiments, the binder may be an agent selected from the group consisting of Advanta-Gel™ P75, Batter Bind® S, Crisp Coat UC, Crisp Film®, Crystal Gum, Crystal TeX™ 627, Crystal Tex™ 644, Crystal Tex™ 648, Elastigel™ 1000J, Encapsul 855, Flojel® 60, Flojel® 65, Flojel® G, Hi-Set® 322, Hi-Set® 377, Hi-Set® C, Hi-Set® CHG, Hylon® V, Hylon® VII, Impression™, K4484, Melojel®, Nadex™ 772, National 0280, National 814, N- TACK®, Purity® 21D, Purity® TF, Superset® LV, Ultra-Set® LT, Dry-Tack® 250, Versa- Sheen™, Baka-Plus™, Baka-Snak®, Capsul®, Capsul® TA, Gel N Melt®, H-50, Hi-Cap™ 100, Hi-Cap™ 200, IF 131, Instant ClearGel®, Instant Pure-Flo, Instant Pure-Flo F, Instant Textaid-A, Instant Textra, National 104, National 1215, National 46, National 1517, National 5730, National 711, National 78-1551, N-Creamer 46, N-Flate, N-Lite™ LP, N-LOK®, N-LOK® 1930, Novation® 4600, Novation® 5600, Novation® 9460, Purity Gum 1773, Purity Gum 2000, Purity Gum 539, Purity Gum BE, Purity® HO, Stir-N-Set® FG, Text-Aid-A®, Textra® Plus, Ultra-Crisp CS, Ultra-Sperse® 2000, Ultra-Sperse® 5, Ultra-Sperse® A, Ultra-Sperse® M, Ultra-Tex 1, UltraTex 2, Ultra-Tex 2000, Ultra-Tex 3, Ultra-Tex 4, AbsorboHP, Amioca, Can-Fil®, Dry-Flo, Hoosier 5, National 150, National 1545, National 6912, National 77-1744, National 912, N-Zorbit® M, Purity® 21, Purity® 5, Purity® 825, Purity® 826, Purity® FC, Target brand tapioca, NU Mould™, Purity® 5S, Clearjel®, Clearjel® S, Colflo® 67, Firm-Tex®, Frigex® w, Hi FLO®, National 1333, National 1457, National 1658, National 4012, National 465, National 740, National Frigex, National Frigex HV, National® 320, Novation® 1600, Novation® 1900, Novation® 2300, Novation® 2600, Novation® 2700, Novation® 3300, Novation® 3600, Novation® 9230, Novation® 9260, Novation® 9270, Novation® 9330, Novation® 9360, Pure-Flo®, Purity® 270, Purity® 4, Purity® 420, Purity® 550, Purity® 660, Purity® 69, Purity® 87, Purity® Cloud, Purity® CSC, Purity® D, Purity® HPC, Purity® W, Thermflo®, Thermtex®, WNA. In still other embodiments, the bromoform bearing core may further comprise: Advanta-Gel™ P75, Batter Bind® S, Crisp Coat UC, Crisp Film®, Crystal Gum, Crystal TeX™ 627, Crystal Tex™ 644, Crystal Tex™ 648, Elastigel™ 1000J, Encapsul 855, Flojel® 60, Flojel® 65, Flojel® G, Hi-Set® 322, Hi-Set® 377, Hi-Set® C, Hi-Set® CHG, Hylon® V, Hylon® VII, Impression™, K4484,Melojel®, Nadex™ 772, National 0280, National 814, N-TACK®, Purity® 21D, Purity® TF, Superset® LV, Ultra-Set® LT, Dry-Tack® 250, Versa-Sheen™, Baka-Plus™, Baka-Snak®, Capsul®, Capsul® TA, Gel N Melt®, H-50, Hi-Cap™ 100, Hi-Cap™ 200, IF 131, Instant ClearGel®, Instant Pure-Flo, Instant Pure-Flo F, Instant Textaid-A, Instant Textra, National 104, National 1215, National 46, National 1517, National 5730, National 711, National 78-1551, N- Creamer 46, N-Flate, N-Lite™ LP, N-LOK®, N-LOK® 1930, Novation® 4600, Novation® 5600, Novation® 9460, Purity Gum 1773, Purity Gum 2000, Purity Gum 539, Purity Gum BE, Purity® HO, Stir-N-Set® FG, Text-Aid-A®, Textra® Plus, Ultra-Crisp CS, Ultra-Sperse® 2000, Ultra- Sperse® 5, Ultra-Sperse® A, Ultra-Sperse® M, Ultra-Tex 1 , Ultra-Tex 2, Ultra-Tex 2000, UltraTex 3, Ultra-Tex 4, AbsorboHP, Amioca, Can-Fil®, Dry -Flo, Hoosier 5, National 150, National 1545, National 6912, National 77-1744, National 912, N-Zorbit® M, Purity® 21, Purity® 5, Purity® 825, Purity® 826, Purity® FC, Target brand tapioca, NU Mould™, Purity® 5S, Clearjel®, Clearjel® S, Colflo® 67, Firm-Tex®, Frigex® w, Hi FLO®, National 1333, National 1457, National 1658, National 4012, National 465, National 740, National Frigex, National Frigex HV, National® 320, Novation® 1600, Novation® 1900, Novation® 2300, Novation® 2600, Novation® 2700, Novation® 3300, Novation® 3600, Novation® 9230, Novation® 9260, Novation® 9270, Novation® 9330, Novation® 9360, Pure-Flo®, Purity® 270, Purity® 4, Purity® 420, Purity® 550, Purity® 660, Purity® 69, Purity® 87, Purity® Cloud, Purity® CSC, Purity® D, Purity® HPC, Purity® W, Thermflo®, Thermtex®, xanthan gum, alginate, a modified polysaccharide, guar gum, acacia gum and combinations thereof.

[0148] In a preferred embodiment, the feed supplement mix comprises between about 0.5% and about 10% binder.

[0149] In a more preferred embodiment, the feed supplement mix comprises between about 1% and about 5% binder.

[0150] In an even more preferred embodiment, the feed supplement mix comprises between about 0.5% and about 3% binder.

[0151] In a preferred embodiment, the feed supplement mix comprises between about 0.5% and about 3% xanthan gum.

[0152] In one embodiment, the feed supplement mix comprises a palatability enhancer. Typically, the palatability enhancer is a flavoring agent such as, but not limited to dry molasses, wet molasses, Qualitech Enhance cantaloupe flavor, or other commercially available animal feed flavor enhancers.Enhanced Feed supplement mix

[0153] In some embodiments, the present technology provides for an enhanced feed supplement mix.

[0154] In one embodiment, the present technology provides for a method of enhancing the bromoform content of a feed supplement mix comprising: a) providing a feed supplement mix; b) treating the feed supplement mix with a peroxide source and optionally, a bromide source or a carboxylic acid; and c) obtaining enhanced feed supplement mix.

[0155] In a preferred embodiment, the starting feed supplement mix comprises between about 30-45% water.

[0156] In a preferred embodiment, the enhanced feed supplement shot comprises between about 20-30%, 30-40%, 35-45%, 40-50%, 45-55%, 50-60%, or 55-65% water.

[0157] In another embodiment, the enhanced feed supplement pre-mix has a bromoform content of more than about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, or about 70% higher than the starting feed supplement pre-mix comprising.

[0158] In a preferred embodiment, the enhanced feed supplement pre-mix has a bromoform content of more than about 50% higher than the feed supplement mix.

[0159] In a preferred embodiment, the peroxide source is sodium percarbonate.

[0160] In a preferred embodiment, the bromide source is sodium bromide.

[0161] In a preferred embodiment, the carboxylic acid is sodium acetate.

[0162] In a preferred embodiment, the biomass is treated only with a peroxide source.

[0163] In an embodiment, the peroxide source is not hydrogen peroxide.

[0164] In an embodiment, the carboxylic acid is not formic acid or a salt or ester thereof.Enriched Feed supplement mix

[0165] In one embodiment, the present technology provides for a method of enriching the bromoform content of a feed supplement mix comprising: a) providing a feed supplement mix, b) treating the feed supplement pre-mix with synthetic or naturally produced bromoform, and c) obtaining enriched feed supplement pre-mix.Feed Supplement Shot

[0166] In one embodiment, the present technology provides a feed supplement shot comprising a feed supplement mix as described herein. In some instances, the feed supplement mix is in a flowable solid form.

[0167] In another embodiment the flowable solid form may be a pellet, a crumble, flake, or powder.

[0168] In a preferred embodiment, the shot is a pellet that is formed from the feed supplement mix without steam treatment.

[0169] The feed supplement shot is typically formed from the feed supplement mix by compression or extrusion. Typically, a pellet mill machine may be used to make the feed supplement shot in pellet form. The feed supplement shot in crumble form may be produced by a crumbier machine. The feed supplement shot in granular form may be produced by a granulatormachine. The types of machinery used to provide the desired physical form of the feed supplement shot are known in the art.

[0170] In one embodiment, the present technology provides for a feed supplement shot comprising at least about 25%, 30%, 35%, 45%, 50%, or 60% moisture.

[0171] In one embodiment, the feed supplement shot comprises at least about 25% moisture. In one embodiment, the feed supplement shot comprises at least about 30% moisture. In one embodiment, the feed supplement shot comprises at least about 35% moisture. In one embodiment, the feed supplement shot comprises at least about 40% moisture. In one embodiment, the feed supplement shot comprises at least about 45% moisture.

[0172] In one embodiment, the present technology provides for a feed supplement shot comprising at least between about 55% moisture. In one embodiment, the present technology provides for a feed supplement shot comprising at least between about 65% moisture. In one embodiment, the feed supplement shot comprises between about 35% and about 55% moisture. In one embodiment, the feed supplement shot comprises between about 40% and about 50% moisture.

[0173] In one embodiment, the feed supplement shot comprises between about 10% and about 40% edible oil. In one embodiment, the feed supplement shot comprises between 12.5% and about 35% edible oil. In one embodiment, the feed supplement shot comprises between about 15% and about 35% edible oil. In one embodiment, the feed supplement shot comprises between 17.5% and 27.5% edible oil. In one embodiment, the feed supplement shot comprises between about 20% and about 30% edible oil. In one embodiment, the feed supplement shot comprises between about 15% and about 35% edible oil.

[0174] In one embodiment, the feed supplement shot comprises between about 20% and about 30% carrier. In one embodiment, the feed supplement shot comprises between about 10% and about 20% carrier.

[0175] In one embodiment, the feed supplement shot comprises wheat middlings as the carrier.

[0176] In one embodiment, the feed supplement shot comprises canola oil as the edible oil.

[0177] In one embodiment, the feed supplement shot comprises between about 0.5% and about 10% binder. In one embodiment, the feed supplement shot comprises between about 1% and about 5% binder. In one embodiment, the feed supplement shot comprises between about 0.5% and about 3% binder.

[0178] In one embodiment, the feed supplement shot comprises between about 0.5% and about 3% xanthan gum.

[0179] In one embodiment, the feed supplement shot is between about 4-13 mm in diameter. In another embodiment, the feed supplement shot is between about 4-10 mm in diameter. In a preferred embodiment, the feed supplement shot is between about 6-8 mm in diameter. In one embodiment, the feed supplement shot has a length between 1.5 and 3 times its diameter.

[0180] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0181] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0182] In a preferred embodiment, the present technology provides for a feed supplement mix comprising:a) between about 70% and about 80% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0183] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0184] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0185] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0186] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0187] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0188] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0189] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 5% and about 50% carrier;c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0190] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0191] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0192] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0193] In one embodiment, the present technology provides for a feed supplement mix comprising:a) between about 60% and about 85% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0194] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0195] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 20% t and about o 30% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0196] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0197] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0198] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 15% palatability enhancer.

[0199] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 15% palatability enhancer.

[0200] In a preferred embodiment, the present technology provides for a feed supplement mix comprising; a) between about 70% and about 80% feed supplement pre-mix; b) between about 5% and about 50% carrier;c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 15% palatability enhancer.

[0201] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 15% palatability enhancer.

[0202] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 15% palatability enhancer.

[0203] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 15% palatability enhancer.

[0204] In one embodiment, the present technology provides for a feed supplement mix comprising:a) between about 50% and about 90% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 15% palatability enhancer.

[0205] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 15% palatability enhancer.

[0206] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 15% palatability enhancer.

[0207] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0208] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0209] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0210] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0211] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 15% and about 40% carrier;c) between about 1% and about 5% binder; and d) between about 0.2% and about to 20% palatability enhancer.

[0212] In a preferred embodiment, the present technology provides for a feed supplement mix comprising; a) between about 70% and about 80% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0213] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0214] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0215] In a preferred embodiment, the present technology provides for a feed supplement mix comprising:a) between about 70% and about 80% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0216] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0217] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0218] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0219] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0220] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0221] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0222] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 20% and about 30% carrier;c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0223] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0224] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 0.5% and about 10% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0225] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 20% palatability enhancer.

[0226] In one embodiment, the present technology provides for a feed supplement mix comprising:a) between about 60% and about 85% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0227] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 5% and about 50% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0228] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0229] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0230] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 15% and about 40% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0231] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 10% and about 20% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0232] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 10% and about 20% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0233] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 10% and about 20% carrier;c) between about 1% and about 5% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0234] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 50% and about 90% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0235] In one embodiment, the present technology provides for a feed supplement mix comprising: a) between about 60% and about 85% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0236] In a preferred embodiment, the present technology provides for a feed supplement mix comprising: a) between about 70% and about 80% feed supplement pre-mix; b) between about 20% and about 30% carrier; c) between about 1% and about 5% binder; and d) between about 0.2% and about 10% palatability enhancer.

[0237] In some embodiments, the feed supplement shot may comprise a binder agent selected from the group consisting of Advanta-Gel™ P75, Batter Bind® S, Crisp Coat UC, CrispFilm®, Crystal Gum, Crystal TeX™ 627, Crystal Tex™ 644, Crystal Tex™ 648, Elastigel™ 1000J, Encapsul 855, Flojel® 60, Flojel® 65, Flojel® G, Hi-Set® 322, Hi-Set® 377, Hi-Set® C, Hi-Set® CHG, Hylon® V, Hylon® VII, Impression™, K4484, Melojel®, Nadex™ 772, National 0280, National 814, N-TACK®, Purity® 21D, Purity® TF, Superset® LV, Ultra-Set® LT, Dry-Tack® 250, Versa-Sheen™, Baka-Plus™, Baka-Snak®, Capsul®, Capsul® TA, Gel N Melt®, H-50, Hi- Cap™ 100, Hi-Cap™ 200, IF 131, Instant ClearGel®, Instant Pure-Flo, Instant Pure-Flo F, Instant Textaid-A, Instant Textra, National 104, National 1215, National 46, National 1517, National 5730, National 711, National 78-1551, N-Creamer 46, N-Flate, N-Lite™ LP, N-LOK®, N-LOK® 1930, Novation® 4600, Novation® 5600, Novation® 9460, Purity Gum 1773, Purity Gum 2000, Purity Gum 539, Purity Gum BE, Purity® HO, Stir-N-Set® FG, Text-Aid-A®, Textra® Plus, Ultra-Crisp CS, Ultra-Sperse® 2000, Ultra-Sperse® 5, Ultra-Sperse® A, Ultra-Sperse® M, Ultra-Tex 1, UltraTex 2, Ultra-Tex 2000, Ultra-Tex 3, Ultra-Tex 4, AbsorboHP, Amioca, Can-Fil®, Dry-Flo, Hoosier 5, National 150, National 1545, National 6912, National 77-1744, National 912, N-Zorbit® M, Purity® 21, Purity® 5, Purity® 825, Purity® 826, Purity® FC, Target brand tapioca, NU Mould™, Purity® 5S, Clearjel®, Clearjel® S, Colflo® 67, Firm-Tex®, Frigex®w, Hi FLO®, National 1333, National 1457, National 1658, National 4012, National 465, National 740, National Frigex, National Frigex HV, National® 320, Novation® 1600, Novation® 1900, Novation® 2300, Novation® 2600, Novation® 2700, Novation® 3300, Novation® 3600, Novation® 9230, Novation® 9260, Novation® 9270, Novation® 9330, Novation® 9360, Pure-Flo®, Purity® 270, Purity® 4, Purity® 420, Purity® 550, Purity® 660, Purity® 69, Purity® 87, Purity® Cloud, Purity® CSC, Purity® D, Purity® HPC, Purity® W, Thermflo®, Thermtex®, WNA.

[0238] In still other embodiments, the bromoform bearing core may further comprise: Advanta-Gel™ P75, Batter Bind® S, Crisp Coat UC, Crisp Film®, Crystal Gum, Crystal TeX™ 627, Crystal Tex™ 644, Crystal Tex™ 648, Elastigel™ 1000J, Encapsul 855, Flojel® 60, Flojel® 65, Flojel® G, Hi-Set® 322, Hi-Set® 377, Hi-Set® C, Hi-Set® CHG, Hylon® V, Hylon® VII, Impression™, K4484, Melojel®, Nadex™ 772, National 0280, National 814, N-TACK®, Purity® 21D, Purity® TF, Superset® LV, Ultra-Set® LT, Dry-Tack® 250, Versa-Sheen™, Baka-Plus™, Baka-Snak®, Capsul®, Capsul® TA, Gel N Melt®, H-50, Hi-Cap™ 100, Hi-Cap™ 200, IF 131, Instant ClearGel®, Instant Pure-Flo, Instant Pure-Flo F, Instant Textaid-A, Instant Textra, National 104, National 1215, National 46, National 1517, National 5730, National 711, National 78-1551, N-Creamer 46, N-Flate, N-Lite™ LP, N-LOK®, N-LOK® 1930, Novation® 4600, Novation®5600, Novation® 9460, Purity Gum 1773, Purity Gum 2000, Purity Gum 539, Purity Gum BE, Purity® HO, Stir-N-Set® FG, Text-Aid-A®, Textra® Plus, Ultra-Crisp CS, Ultra-Sperse® 2000, Ultra- Sperse® 5, Ultra-Sperse® A, Ultra-Sperse® M, Ultra-Tex 1, Ultra-Tex 2, Ultra-Tex 2000, Ultra-Tex 3, Ultra-Tex 4, AbsorboHP, Amioca, Can-Fil®, Dry-Flo, Hoosier 5, National 150, National 1545, National 6912, National 77-1744, National 912, N-Zorbit® M, Purity® 21, Purity® 5, Purity® 825, Purity® 826, Purity® FC, Target brand tapioca, NU Mould™, Purity® 5S, Clearjel®, Cleaijel® S, Colflo® 67, Firm-Tex®, Frigex® w, Hi FLO®, National 1333, National 1457, National 1658, National 4012, National 465, National 740, National Frigex, National Frigex HV, National® 320, Novation® 1600, Novation® 1900, Novation® 2300, Novation® 2600, Novation® 2700, Novation® 3300, Novation® 3600, Novation® 9230, Novation® 9260, Novation® 9270, Novation® 9330, Novation® 9360, Pure-Flo®, Purity® 270, Purity® 4, Purity® 420, Purity® 550, Purity® 660, Purity® 69, Purity® 87, Purity® Cloud, Purity® CSC, Purity® D, Purity® HPC, Purity® W, Thermflo®, Thermtex®, xanthan gum, alginate, a modified polysaccharide, guar gum, acacia gum and combinations thereof.

[0239] In one embodiment, the present technology provides for a feed supplement mix or shot made from the ingredients, percentage composition and in the ratios of those ingredients as shown in Tables 1 and 2 wherein the ratios may vary by about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10%.

[0240] In one embodiment, the present technology provides for a feed supplement mix or shot made from the ingredients, percentage compositions and in the ratios of those ingredients as shown in Tables 1 and 2 wherein the ratios may vary by between about 1% and about 5%, between about 2% and about 6%, between about 3% and about 7%, between bout 4% and about 8%, between about 5% and about 9%, or between about 6% and about 10%.

[0241] In one embodiment, the present technology provides for a feed supplement mix or shot comprising the ratios of bromoform, iodine, and moisture as shown in Tables 1 and 2 wherein the ratios may vary by about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10%.

[0242] In one embodiment, the present technology provides for a feed supplement mix or shot comprising the ratios of bromoform, iodine, and moisture as s shown in Tables 1 and 2 wherein the ratios may vary by between about 1% and about 5%, 2% and about 6%, 3% and about 7%, 4% and about 8%, 5% and about 9%, or 6% and about 10%.

[0243] In one embodiment, the present technology provides for a feed supplement premix, mix or shot having a bromoform to iodine ratio of at least 5 wt / wt.

[0244] In one embodiment, the present technology provides for a feed supplement premix, mix or shot having a bromoform to iodine ratio of at least 9 wt / wt.

[0245] In one embodiment, the present technology provides for a kit comprising a feed supplement pre-mix, mix, or shot as described herein, a iodine feed supplement, and instructions for feeding a combination of the feed supplement pre-mix, mix, or shot as described herein along with the iodine feed supplement.

[0246] In some embodiments, the present technology provides for a Asparagopsis based feed supplement pre-mix that shows less than 1%, 2%, 3%, 4% or 5% bromoform loss when stored without refrigeration for over 10 weeks, 15 weeks, 20 weeks or 25 weeks.

[0247] In another embodiment, the present technology provides for a bromoform producing yeast-based feed supplement mix that shows less than 1%, 2%, 3%, 4% or 5% bromoform loss when stored without refrigeration for over 10 weeks, 15 weeks, 20 weeks or 25 weeks.

[0248] In another embodiment, the present technology provides for a bromoform supplemented based feed supplement shot that shows less than 1%, 2%, 3%, 4% or 5% bromoform loss when stored without refrigeration for over 10 weeks, 15 weeks, 20 weeks or 25 weeks.

[0249] some embodiments, the present technology provides for a Asparagopsis based feed supplement pre-mix that shows less than 5000 cfu / g of aerobic plate count when stored without refrigeration for over 10 weeks, 15 weeks, 20 weeks or 25 weeks.

[0250] In another embodiment, the present technology provides for a bromoform producing yeast-based feed supplement mix that shows less than 5000 cfu / g of aerobic plate count when stored without refrigeration for over 10 weeks, 15 weeks, 20 weeks or 25 weeks.

[0251] In another embodiment, the present technology provides for a bromoform supplemented biomass-based feed supplement shot that shows less than 5000 cfu / g of aerobic plate count when stored without refrigeration for over 10 weeks, 15 weeks, 20 weeks or 25 weeks.

[0252] In still another embodiment, the present technology provides for substitution of a bromoform producing yeast-based feed supplement pre-mix, feed supplement mix or shot having the stability properties described herein for the corresponding Asparagopsis based formulations.

[0253] In still another embodiment, the present technology provides for substitution of a bromoform producing organism-based feed supplement pre-mix, feed supplement mix or shot having the stability properties described herein for the corresponding Asparagopsis based formulations.

[0254] In one embodiment, the bromoform enriched biomass-based feed supplement mix comprises about Img / g to 20 mg / g bromoform.

[0255] In a preferred embodiment, the bromoform enriched biomass-based feed supplement mix comprises about 5mg / g to 15 mg / g bromoform.

[0256] In one embodiment, the bromoform producing yeast-based feed supplement mix comprises about Img / g to 20 mg / g bromoform.

[0257] In a preferred embodiment, the bromoform producing yeast-based feed supplement mix comprises about 5mg / g to 15 mg / g bromoform.

[0258] In one embodiment, the Asparagopsis ssp. -based feed supplement mix comprises about Img / g to 20 mg / g bromoform.Enhanced Feed Supplement Shot

[0259] In some embodiments, the present technology provides for enhanced feed supplement pre-mixes, enhanced feed supplement mixes, and enhanced feed supplement shot.

[0260] In one embodiment, the present technology provides for a method of enhancing the bromoform content of a feed supplement shot comprising: a) providing a feed supplement shot; b) treating the feed supplement shot with a peroxide source and optionally, a bromide source or a carboxylic acid; andc) obtaining enhanced feed supplement shot.

[0261] In a preferred embodiment, the starting feed supplement shot comprises between about 35 and about 45% water.

[0262] In a preferred embodiment, the starting feed supplement shot comprises between about 20-30%, 25-35%, 30-40%, 35-45%, 40-50%, 45-55%, 50-60%, 55-65%, or 60-70% water.

[0263] In another embodiment, the enhanced feed supplement pre-mix has a bromoform content of more than 10%, 20%, 30%, 40%, 50%, 60%, or 70% higher than the starting feed supplement pre-mix comprising.

[0264] In a preferred embodiment, the enhanced feed supplement pre-mix has a bromoform content of more than 50% higher than the feed supplement pre-mix.

[0265] In a preferred embodiment, the peroxide source is sodium percarbonate.

[0266] In a preferred embodiment, the bromide source is sodium bromide.

[0267] In a preferred embodiment, the carboxylic acid is sodium acetate.

[0268] In a preferred embodiment, the biomass is treated only with a peroxide source.

[0269] In an embodiment, the peroxide source is not hydrogen peroxide.

[0270] In an embodiment, the carboxylic acid is not formic acid or a salt or ester thereof.Enriched Feed supplement shot

[0271] In one embodiment, the present technology provides for a method of enriching the bromoform content of a feed supplement shot comprising: a) providing a feed supplement shot, b) treating the feed supplement shot with synthetically or naturally produced bromoform, and c) obtaining enriched feed supplement shot.

[0272] Asparagopsis feed supplement shot pellets maintain the bromoform stability and long shelflife of the feed supplement mix base product and are a palatable & user friendly method for delivery of the naturally occurring bioactive compounds within Asparagopsis Taxiformis / Armaia into the diet of ruminant livestock. Inclusion rates depend on the concentration of bromoform (the primary bioactive compound) within the final Asparagopsis feed supplement shot pellets product, the type and life-stage of the ruminant livestock, and the emission reduction targets of the user. Once a target inclusion rate is selected for a particular livestock application, Asparagopsis feed supplement shot pellets are then added to (or top-dressed onto) the Total Mixed Ration (TMR) of the livestock and consumed as part of the animals normal daily feeding activities. The bioactive compounds then inhibit the methanogenic metabolic pathways within the rumen of the animal to prevent methanogenesis and reduce rumen related methane emissions by 50 - 90%

[0273] In one embodiment, the feed supplement pre-mix is shelf stable under typical farm storage conditions with respect to bromoform for at least 6 month.

[0274] It is understood that the present technology can be applied to any biomass derived from a bromoform producing organism and that in any embodiment or example disclosed herein, Asparagopsis spp. biomass can be interchanged with bromoform producing yeast, bacteria, other algae, fungi, or plant.

[0275] Table 1 : Examples of Asparagopsis feed supplement shot pellets Formulations made from dewatered Asparagopsis comprising 75% moisture. The example includes the ingredients used to make the recipes and the calculated final composition of feed supplement shot pellets, together with measured parameters and results from stability tests. Stability tests examined bromoform retention through pelletization and storage (4 weeks at room temperature) of feed supplement shot pellets.

[0276] Table 2: Additional examples of Asparagopsis feed supplement shot pelletsFormulations made from dewatered Asparagopsis comprising 75% moisture.

[0277] Table 3: Average Bromoform concentration in feed supplement-pre-mix (mg / g) fromExample 1.

[0278] Table 4: Average Bromoform retention in feed suppl ement-pre-mix (% from bromoform in dewatered biomass) from Example 1.

[0279] Table 5: Average bromoform concentration from week 0 to week 22 from Example 2.

[0280] Table 6: Average pathogen count from week 11 and week 22 from the compositions ofExample 2

[0281] Table 7: Bromoform Retention through pelletization for Asparagopsis feed supplement shot pellets from example 3.

[0282] Table 8: Bromoform Retention through storage for Asparagopsis feed supplement shot pellets from example 3.

[0283] Table 9: Formulation of pellets used for storage stability test in example 4. FSP = FeedSupplement Pre-Mix

[0284] Table 10: Table Formulation of control pellets used for storage stability test in example

[0285] Table 11 : Average bromoform concentration in Brominata Pellets from week 0 to week8 from example 4

[0286] Table 12: Microbial test results for Brominata and Control Pellets from week 0 to week8. Due to high pathogen levels measured by week 4, control pellets were discarded and not sampled at week 8 from example 4

[0287] Table 13 : Bromine, iodine, and bromoform content measured in Brominata pellets 0 and4 weeks after pelletization from example 4.

[0288] Table 14: Exemplary inclusion rates for Asparagopsis feed supplement shot pellets based on bromoform concentration in pellets for at least a 50% reduction in methane production.

[0289] Table 15: Formulas and final composition of Asparagopsis feed supplement shot andControl pelletsDM = Dry MatterTable 16: Feed supplement pre-mix formulations tested for post-harvest bromoform enhancement from Example 9

[0290] Table 17: Brominata Feed Supplement (BFS) Pre-Mix formulations tested for postharvest bromoform enhancement from Example 10EXAMPLESExample 1. Feed Supplement Pre-Mix

[0291] Example 1 shows the preparation of the feed supplement pre-mix according to the recipes shown in Tables 1 and 2. Its stability with respect to bromoform, is shown in Figure 1 and2 and Tables 3 and 4.

[0292] Asparagopsis taxiformis tetrasporophyte biomass was dewatered by centrifugation to a 75% residual moisture content. Samples of this dewatered biomass were freeze dried to provide samples for bromoform content. The dewatered biomass having 75% residual moisture content was mixed with canola oil in three different batches corresponding to 80%:20%, 65%:35% and 50%:50% dewatered biomass to oil content ratios and the batches were individually homogenized using a mastification juicer to obtain batches of feed supplement pre-mix compositions. The stability of the feed supplement pre-mixes exemplified here are shown in Figures 1 and 2 and Tables3 and 4.Example 2. Feed Supplement Pre-Mix Stability Test

[0293] Example 2 shows the extended bromoform stability properties and pathogen resistance of the feed supplement pre-mix over a period of 22 weeks under typical farm conditions for a composition comprising an initial 25% dry matter and 32% oil. The bromoform stability is shown in Figure 3 and Table 5. The pathogen analysis is shown in Table 6.

[0294] To examine the stability of Asparagopsis-based feed supplement pre-mix, a storage stability test was conducted by Blue Ocean Barns, Erendira, MX, to track the change in bromoform concentration within the product over the course of five months. The results indicatedno loss of bromoform in Asparagopsis supplement pre-mix stored under farm conditions for five months (22 weeks).

[0295] For the storage stability test, 15kg of Asparagopsis taxiformis tetrasporophyte algae was dewatered using a screw press equipped with oil injection nozzles, to reach dry matter content (DM%) of 25%, and canola oil content of 32% to yield the desired feed supplement premix. Oil addition was performed during the dewatering process using a screw press equipped with oil injection nozzles where biomass was masticated with oil to create the overall 32% oil mixture. Oil percentage was further verified based on DM% measurements.

[0296] The feed supplement pre-mix was sampled for bromoform and distributed into two 1 -gallon containers that were sealed and only opened for sampling throughout the storage stability test. All containers were stored in a non-temperature controlled covered area. The containers were opened at week 2, 4, 8, 11, 17, and 22 to measure bromoform retention over time. Bromoform was sampled from the feed supplement pre-mix after it had been well mixed at each time point. Pathogen analysis was also sampled at week 11, 17, and 22.

[0297] Bromoform Concentration (mg'g): Samples were taken in a 50ml conical tube in Erendira, MX and then sent to the Blue Ocean Barns Analytical Lab in San Diego, CA. Once received in San Diego, the samples were incubated at 60°C for 4 days in order to speed up bromoform extraction into the oil in the pesto. For each bromoform sample, 2 g of the feed supplement pre-mix was placed in a 2 ml Eppendorf tube and centrifuged for 10 minutes at 12000 RPM to separate the oil. Oil samples (25 mg) were taken from each sample and then extracted and measured using a Gas Chromatography -Mass Spectrometer. Samples were run on the GCMS within two hours of being incubated.

[0298] Pathogen Detection: To determine the presence of pathogens, a sample (50g) was taken from each bucket at weeks 11 and 22 and sent to Midwest Laboratories for analysis. The lab tested for the presence and concentration of E. coli (cfu / g), total coliforms (cfu / g), Salmonella (organisms / 25g), Staphylococcus aureus (cfu / g), Yeast (cfu / g), mold count (cfu / g), and Aerobic Plate Count (cfu / g).

[0299] Results'. The initial bromoform concentration within the feed supplement pre-mix was 7.28 + / - 0.20mg / g. The average bromoform concentration at 22 weeks for the feed supplementpre-mix was 7.63 + / - 0.17 mg / g, Data showed that there was no statistically significant bromoform loss during the 22-week storage of the feed supplement pre-mix (Fig. 3, Table 5).

[0300] Samples were taken at the end of the 11 and 22 weeks to measure pathogen presence within the feed supplement pre-mix. The results of the pathogen test found no measurable amounts ofE. coli, Total coliforms, Salmonella, Staphylococcus aureus, Yeast, or Mold (Table 6).

[0301] Conclusions: The feed supplement pre-mix was stable over a 22-week period when stored under farm conditions, with no decrease in bromoform over that time period. No specific pathogens were detected after 22 weeks and there was no increase in the aerobic plate count at the end of the trial. The aerobic plate count was significantly under the 5000cfu / g specification at all times. Feed Supplement Pre-Mix produced from pesto stored for over three months have been included in several months-long feeding trials with cows, showing no signs of pellet refusal or negative effect on cattle health.Example 3. Feed Supplement Shot and Bromoform Stabdity

[0302] Example 3 shows the preparation of the feed supplement shot pellets from various formulations (recipes) of feed supplement pre-mix and feed supplement mix. These formulations (recipes) are illustrated in Tables 1 and 2. Table 1 also shows the final composition parameters and bromoform retention upon room temperature storage. Table 2 illustrates alternative formulations (recipes), as well as the final feed supplement shot pellet compositions that are the subject of the present technology. The bromoform stability of these compositions is shown in Figures 4 and 5 and Tables 7 and 8.Preparation of Feed Supplement Mix Based Pellets

[0303] Live material: Asparagopsis biomass was harvested from a production tank at Blue Ocean Barns’ production facility in Kona, Hawaii.

[0304] Dewatering: Freshly harvested Asparagopsis biomass was dewatered by centrifugation to 75% final moisture content. Bromoform samples were taken from the freeze dried dewatered biomass, and measured at Blue Ocean Barns’ GCMS facility to represent initial bromoform levels in biomass.

[0305] Feed supplement pre-mix: Dewatered biomass was mixed with canola oil at final oil concentration of 30%. Oil and biomass were homogenized using a masticating juicer to get a homogenic Asparagopsis-oil mixture. Bromoform samples were taken from feed supplement premix and measured at Bigelow Analytical Services’ GCMS facility to represent initial bromoform levels in the feed supplement mix.

[0306] Feed supplement mix: Feed supplement mix mixtures for feed supplement shot pellets were made following the formulations shown in Example 1 (Table 1). To make IKg of feed supplement shot pellet formulation #1. 750g of Asparagopsis feed supplement pre-mix, 237.5g of carrier (wheat middling), 2.5 g of palatability enhancer (Qualitech Accelerate), and 10 g of binder (xanthan gum), were mixed in a feed processor to make a homogenous feed supplement mix. To make IKg of feed supplement shot pellet formulation #2. 850g of Asparagopsis feed supplement pre-mix, 137.5g of carrier (wheat middling), 2.5 g of palatability enhancer (Qualitech Accelerate), and 10 g of binder (xanthan gum), were mixed in a feed processor to make a homogenous feed supplement mix mixture.

[0307] Pelletization: Feed supplement mixes were run through a cold press pellet mill die to create 6 mm diameter pellets. Bromoform samples were taken from Pellets and measured at Bigelow Analytical Services’ GCMS facility to represent initial bromoform levels in the pellets.

[0308] Bromoform Retention through pelletization: Bromoform retention through pelletization was calculated as follows:

[0309] Bromoform retention = bromoform in pellets / (mix inclusion in pellets X bromoform in mix)

[0310] Bromoform results showed no significant bromoform loss through pelletization. On average, bromoform retention was better for pellets made with 85% feed supplement mix base as shown in graphical and tabular form in Figures 4 and 5 and Tables 7 and 8.Example 4. Feed Supplement Shot and Bromoform Extended Stability Test

[0311] Example 4 shows the bromoform stability and pathogen resistance of an Asparagopsis-based feed supplement shot pellet composition that is the subject of the present technology. The composition of this feed supplement shot is shown in Table 9 and the kelp-basedcontrol pellets in Table 10. Table 11 and Figure 6 shows the bromoform stability over an 8-week period of the Asparagopsis based composition, while Table 12 shows the pathogen resistance versus the kelp-based pellets. Table 13 additionally shows bromoform and iodine content over a 4-week period.

[0312] To evaluate the stability of Asparagopsis based feed supplement shot, Blue Ocean Barns in Erendira, MX, conducted a three-month storage stability test, the study monitored changes in bromoform concentration and performed microbial assessments of the product. Over a two- month period, the data revealed no significant bromoform loss and maintained acceptable microbial levels. In contrast, control pellets - formulated similarly to the Asparagopsis based feed supplement shot but substituting Asparagopsis biomass with kelp biomass - exhibited significant bacterial contamination and molding within the first two weeks, underscoring the distinctive stability of Asparagopsis based feed supplement shot.

[0313] Asparagopsis biomass was harvested from a production tank at Blue Ocean Barns’ production facility in Erendira, Baja California, Mexico. The freshly harvested Asparagopsis biomass was dewatered by a screw press to 73% final moisture content (DM = 27%). Bromoform samples were taken from the dewatered biomass, freeze dried and measured at Blue Ocean Barns’ GCMS facility to represent initial bromoform levels in biomass. The dewatered biomass was mixed with canola oil at final oil concentration of 30% to get a homogenic feed supplement pre-mix.

[0314] Feed supplement mix formulations for pelleting were made following the formulations shown in Tables 8 and 9. Mash mixtures were run through a cold press pellet mill die to create 6 mm diameter pellets and Bromoform samples were taken from the pellets and measured at Blue Ocean Bams GCMS facility to represent initial bromoform levels in the pellets.

[0315] The pellets were packaged in heat-sealed PE bags, lOKg pellets in each bag, 3 bags of Asparagopsis pellets and 2 bags of control pellets. All bags were stored in the harvest hut, which is covered but not temperature controlled.

[0316] Bromoform sampling was conducted from Asparagopsis pellet bags at weeks 2, 4, and 8. Each bag was only opened during sampling. To avoid potential artifacts caused by variability within the bags, bromoform samples were taken from multiple locations in each bag.

[0317] Samples for microbial tests (Aerobic plate count, E. Coli, Total coliforms, Salmonella, Staphylococcus aureus, Yeast, and Mold count) were taken from Asparagopsis pellets and control pellets at weeks 0, 2, 4, and 8 to monitor pellet stability

[0318] Samples for ICP-MS analysis of bromine and iodine were taken from Asparagopsis pellets at weeks 0 and 4.

[0319] Bromoform Concentration (mg / g): Samples were taken in a 50ml conical tube in Erendira, MX and then sent to the BOB Analytical Lab in San Diego, CA. For each bromoform sample, 2 g of Asparagopsis pellets were placed in a 2 ml Eppendorf tube and centrifuged for 10 minutes at 12000 RPM to separate the oil from the biomass. Oil samples (25 mg) were taken from each sample and then extracted and measured using a Gas Chromatography-Mass Spectrometer. Samples were run on the GCMS within two hours of being incubated.

[0320] Pathogen Detection: To determine the presence of pathogens, a sample (50g) was taken from each bag at weeks 0, 2, 4, and 8 and sent to Midwest Laboratories for analysis. The lab tested for the presence and concentration of E. coli (cfu / g), total coliforms (cfu / g), Salmonella (organisms / 25g), Staphylococcus aureus (cfu / g), Yeast (cfu / g), mold count (cfu / g), and Aerobic Plate Count (cfu / g).

[0321] Bromine and iodine content: To determine bromine and iodine concentrations in Brominata pellets, a sample (50g) was taken from each bag at weeks 0 and 4 and sent to Eurofins Laboratories for Inductively Coupled Plasma Mass Spectrometry (ICP-MS) analysis.

[0322] The initial bromoform concentration in Brominata Pellets was 4.78 ± 0.07 mg / g. After 8 weeks of storage, the average concentration decreased slightly to 4.52 ± 0.09 mg / g (Fig. 6, Table 11). However, Mixed Linear Model Regression analysis indicated no statistically significant reduction in bromoform levels over time (p = 0.991). Microbial testing revealed no detectable presence of E. coli, Total coliforms, Salmonella, Staphylococcus aureus, yeast, or mold, while aerobic plate counts remained within acceptable limits (Table 12). Asparagopsis pellets demonstrated stability over a 8-week period, with no significant loss of bromoform, and acceptable microbial counts during this time. No specific pathogens were detected after 8 weeks and there was no increase in the aerobic plate count at the end of the trial. The aerobic plate count was acceptable for all conditions as shown in Table 11.Example 5. Inclusion of Asparagopsis feed supplement shot pellets in dairy cow diet.

[0323] Example 5 shows representative target feed inclusion rates of the feed supplement mix or shot compositions of the present technology in order to achieve at least a 50% reduction in methane emissions from dairy cows based on the bromoform content of the feed supplement mix or shot as shown in Table 14.

[0324] Asparagopsis feed supplement shot pellets may be included in cattle feed to reduce methane emissions by 50 - 90%. Pellet inclusion rate is calculated based on the active ingredient (bromoform) concentration in pellets to achieve bromoform inclusion rate of 16 mg / KgDMI (Table 14).Example 6. Asparagopsis Feed Supplement Shot Mold Stability Test

[0325] Example 6 shows the shelf stability of an example of the feed supplement shot of the present technology. Stability testing of a typical example of the feed supplement shot of the present technology has shown that the shot is shelf stable with respect to molding for a period of at least 2 weeks at room temperature when stored in a closed container. In one embodiment, the feed supplement shot is shelf stable with respect to molds for at least 3, 4, 6, and 8 weeks at room temperature when stored in a closed container. Figure 7 shows the visual difference between control kelp-based pellets and the Asparagopsis based pellets after storage for 2 weeks at room temperature.

[0326] Asparagopsis feed supplement shot pellets were compared to control pellets with similar oil and water content that did not consist of Asparagopsis. Asparagopsis feed supplement shot and control pellets were made following the formulas in Table 15. In the control pellets, Asparagopsis feed supplement shot was replaced with dried Kelp powder added with canola oil and water to reach final oil and water inclusion similar to Asparagopsis feed supplement shot pellets. Both pellets were stored together in a Ziplock bag at room temperature for 14 days. About 7 days through storage mold was evidenced on control pellets. At the end of the 14 days trial, control pellets were heavily molded while no mold was apparent on Asparagopsis feed supplement shot pellets (Figure 7).Example 7. Yeast-based feed supplement pre-mix, mix and shot.

[0327] Example 7 discloses a method of making a feed supplement shot with a bromoform producing yeast biomass instead of bromoform producing algal biomass.

[0328] Bromoform producing yeast is grown in a fermentation vessel resulting in a slurry of the biomass in the aqueous culture medium, wherein the biomass comprises 10 mg / g to 50 mg / g bromoform on a per dry weight basis. In some cases, the culture medium may be supplemented with a canola oil overlay comprising between 0 to 35% of the total cultivation volume. The slurry is harvested and the moisture content of the slurry is adjusted to about 45% by either adding water or removing water via centrifugation or decantation.

[0329] Canola oil is added to adjust the total oil content to about 30% by volume, and the mixture is homogenized using a screw press to yield a yeast-based feed supplement pre-mix. Bromoform content of the yeast-based feed supplement pre-mix is determined by GCMS.

[0330] Yeast-based feed supplement mix is prepared by mixing 750 g of the yeast-based feed supplement pre-mix, 237.5 g of a carrier, such as wheat middling, 2.5 g of a palatability enhancer such as Qualitech Accelerate, and 10 g of a binder, such as xanthan gum. The mixing is performed in a dough mixer until a homogeneous mass is achieved.

[0331] Another formulation of yeast-based feed supplement mix is prepared by mixing 850 g of the yeast-based feed supplement pre-mix, 137.5 g of a carrier, such as wheat middling, 2.5 g of a palatability enhancer such as Qualitech Accelerate, and 10 g of a binder, such as xanthan gum. The mixing is performed in a dough mixer until a homogeneous mass is achieved.

[0332] Either yeast-based feed supplement mix is pelletized by running the mix through a cold press mill die to create 6mm diameter pellets. Bromoform content of the pelletized yeastbased feed supplement pre-mix (yeast-based feed supplement shot) is determined by GCMS.Example 8. Bromoform enriched feed supplement pre-mix, mix and shot.

[0333] Example 8 discloses how the feed supplement shot may be enriched in bromoform using synthetic or naturally derived bromoform.

[0334] Bromoform producing biomass is grown in a culture vessel resulting in a slurry of the biomass in the aqueous culture medium. In some cases, the culture medium may besupplemented with a canola oil overlay comprising between 0 to 35% of the total cultivation volume. The slurry is harvested and the moisture content of the slurry is adjusted to about 20% to30% by either adding water or removing water via centrifugation or decantation.

[0335] Canola oil is added to adjust the total oil content to about 30% by volume, and the mixture is homogenized using a screw press to yield a biomass-based feed supplement pre-mix. Bromoform content of the biomass-based feed supplement pre-mix is determined by GCMS.

[0336] The bromoform concentration of the bromoform supplemented biomass-based feed supplement pre-mix is adjusted to a desired level by the addition of synthetic or organically derived bromoform to between about Img / g to 20 mg / g bromoform.

[0337] In a preferred embodiment, the Asparagopsis ssp. -based feed supplement mix comprises about 5mg / g to 15 mg / g bromoform.

[0338] Biomass-based feed supplement mix is prepared by mixing 750 g of the enriched biomass-based feed supplement pre-mix, 237.5 g of a carrier, such as wheat middling, 2.5 g of a palatability enhancer such as Qualitech Accelerate, and 10 g of a binder, such as xanthan gum. The mixing is performed in a dough mixer until a homogeneous mass is achieved.

[0339] Another formulation of biomass-based feed supplement mix is prepared by mixing 850 g of the enriched biomass-based feed supplement pre-mix, 137.5 g of a carrier, such as wheat middling, 2.5 g of a palatability enhancer such as Qualitech Accelerate, and 10 g of a binder, such as xanthan gum. The mixing is performed in a dough mixer until a homogeneous mass is achieved.

[0340] Either biomass-based feed supplement mix is pelletized by running the mix through a cold press mill die to create 6mm diameter pellets. Bromoform content of the pelletized biomass-based feed supplement pre-mix (biomass-based feed supplement shot) is determined by GCMS.

[0341] The resulting supplemented mix or shot shows less than 2% bromoform loss even after storing in a sealed light proof container for at least 2 months.Example 9. Bromoform enhanced feed supplement pre-mix, mix and shot.

[0342] Example 9 describes the production of enhanced feed supplement pre-mix from feed supplement pre-mix. Asparagopsis taxiformis biomass and Canola oil was added with bromoform enhancement ingredients to facilitate the post-harvest production of bromoform during early storage (4 weeks). Inclusion of sodium percarbonate whether with either acetate or formate resulted in up to 71% increase in bromoform compared to initial bromoform levels, and 53% increase in comparison to non-enhanced feed supplement pre-mix.

[0343] Asparagopsis taxiformis biomass-based feed supplement pre-mix was produced from A. taxiformis biomass cultivated at Blue Ocean Bams San Diego R&D facility. Initial bromoform levels in biomass was measured as 54 mg / gDW.

[0344] The biomass was dewatered using a Panda 3200 RPM Portable Spin Dryer to reach30% dry matter (DM%) and mixed with canola oil to create 30% oil / biomass feed supplement premix. The DM% was measured by drying at 65 deg. C for 48 hours to confirm initial DM% and post-mixing oil inclusion.

[0345] The feed supplement pre-mix was treated with various combinations of sodium acetate, sodium formate, sodium bromide, and sodium percarbonate as shown in Table 16.

[0346] All the control and enhanced feed supplement pre-mix formulations other than formulation #10 were placed in black mylar bags, 2 bags per formulation, and stored at room temperature. 2 samples from formulation #10 were placed in mylar bags and stored in a -80 deg. C deep freezer.

[0347] Bromoform was sampled from each bag at weeks 0 and 4. In week 0, the control and enhanced feed supplement pre-mix samples were incubated at 60 deg. C for 4 days before GCMS analysis to allow bromoform to be extracted into the oil phase.

[0348] For each bromoform sample, 2 g of the control and enhanced feed supplement premix was placed in a 2 ml Eppendorf tube and centrifuged for 10 minutes at 6000 RPM to separate the oil. Oil samples (25 mg) were taken from each tube and then extracted in MeOH and measured using a Gas Chromatography -Mass Spectrometer. Samples were run on the GCMS within two hours of being incubated.

[0349] All supplemented formulations (formulations # 2-9) showed an increase of 11% or more in bromoform compared to non-enhanced feed supplement pre-mix. Formulations which included sodium percarbonate addition (formulations # 2-5) showed higher bromoform increase, resulting in 38-53% higher concentrations compared to non-enhanced feed supplement pre-mix after 4 weeks.Example 10. Bromoform enhanced feed supplement pre -mix, mix and shot.

[0350] Example 10 describes the production of enhanced feed supplement pre-mix from feed supplement pre-mix. Asparagopsis taxiformis biomass and Canola oil was added with bromoform enhancement ingredients to facilitate the post-harvest production of bromoform during early storage (4 weeks). Inclusion of sodium percarbonate whether with or without sodium acetate resulted in up to 49% increase in bromoform compared to initial bromoform levels, and 61% increase in comparison to non-enhanced feed supplement pre-mix.

[0351] Asparagopsis taxiformis biomass feed supplement pre-mix was produced from A. taxiformis biomass cultivated at Blue Ocean Bams San Diego R&D facility. Initial bromoform levels in biomass was measured as 26 mg / gDW.

[0352] The biomass was dewatered using a Panda 3200 RPM Portable Spin Dryer to reach 30% dry matter (DM%) and mixed with canola oil to create 30% oil / biomass feed supplement premix. The DM% was measured by drying at 65 deg. C for 48 hours to confirm initial DM% and post-mixing oil inclusion.

[0353] The feed supplement pre-mix was added with various combinations of sodium acetate and sodium percarbonate as shown in Table 17.

[0354] All the feed supplement pre-mix formulations other than formulation #10 were placed in 50 ml centrifuge tubes, 3 tubes per formulation, and stored at room temperature. 3 samples from formulation #10 were placed in 50 ml centrifuge tubes and stored in a -80 deg. C deep freezer.

[0355] Bromoform was sampled from each bag at weeks 0 and 4. In week 0, control and enhanced feed supplement pre-mix samples were incubated at 60 deg. for 4 days before GCMS analysis to allow bromoform to be extracted into the oil phase.

[0356] For each bromoform sample, 2 g of either the control or the enhanced feed supplement pre-mix was placed in a 2 ml Eppendorf tube and centrifuged for 10 minutes at 6000 RPM to separate the oil. Oil samples (25 mg) were taken from each tube and then extracted in MeOH and measured using a Gas Chromatography-Mass Spectrometer. Samples were run on the GCMS within two hours of being incubated.

[0357] All supplemented formulations (formulations # 2-9) showed an increase of 28% or more in bromoform compared to non-supplemented Brominata Feed Supplement Pre-Mix. Sodium percarbonate addition at 2% level, with or without sodium acetate, (formulations # 3 and #8) showed bromoform increase of 52-61% compared to non-supplemented Brominata Feed Supplement Pre-Mix after 4 weeks.

[0358] Any element of any embodiment may be used in any embodiment. Although the invention has been described with reference to specific embodiments, it will be understood by those skilled in the art that various changes may be made, and equivalents may be substituted for elements thereof without departing from the true spirit and scope of the invention. In addition, modifications may be made without departing from the essential teachings of the invention. Identification of equivalent compositions, methods and kits are well within the skill of the ordinary practitioner and would require no more than routine experimentation, in light of the teachings of the present disclosure. Practice of the disclosure will be still more fully understood from the following examples, which are presented herein for illustration only and should not be construed as limiting the disclosure in any way.

[0359] All references cited in this specification, and their references, are incorporated by reference herein in their entirety where appropriate for teachings of additional or alternative details, features, and / or technical background.

[0360] While the disclosure has been particularly shown and described with reference to particular embodiments, it will be appreciated that variations of the above-disclosed and other features and functions, or alternatives thereof, may be desirably combined into many other different systems or applications. Also, that various presently unforeseen or unanticipated alternatives, modifications, variations or improvements therein may be subsequently made by those skilled in the art which are also intended to be encompassed by the following claims.

Claims

CLAIMS:

1. A feed supplement pre-mix comprising: a) between about 50% and about 90% dewatered Asparagopsis spp. biomass; and b) between about 15% and about 50% edible oil, wherein the dewatered Asparagopsis spp. has a moisture content of between about 50% and about 80%.

2. The feed supplement pre-mix of claim 1, wherein the dewatered Asparagopsis spp. biomass has a moisture content of between about 60% and about 80%.

3. The feed supplement pre-mix of claim 1, wherein the dewatered Asparagopsis spp. biomass has a moisture content of between about 65% and about 75%.

4. The feed supplement pre-mix of claim 3, comprising between about 60% and about 80% dewatered Asparagopsis spp biomass.

5. The feed supplement pre-mix of claim 3, comprising between about 65% and about 75% dewatered Asparagopsis spp biomass.

6. The feed supplement pre-mix of any one of claims 1 to 5, further comprising between about 0.1% and about 5% xanthan gum.

7. A feed supplement mix comprising: a) between about 50% and about 90% of a feed supplement pre-mix, b) between about 5% and about 50% carrier, c) between about 0.5% and about 10% binder, and d) between about 0.2% and about 20% palatability enhancer, wherein the feed supplement pre-mix comprises between about 50% and about 90% dewatered Asparagopsis biomass and between about 50% and about 10% edible oil; andwherein the dewatered Asparagopsis biomass comprises between about 50% and about 80% residual moisture.

8. The feed supplement mix of claim 7, wherein the feed supplement mix comprises between about 60% and about 85% of the feed supplement pre-mix.

9. The feed supplement mix of claim 7, wherein the feed supplement mix comprises between about 70% and about 80% of the feed supplement pre-mix.

10. The feed supplement mix of claim 7, wherein the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

11. The feed supplement mix of claim 7, wherein the feed supplement mix comprises between about 60% and about 85% of the feed supplement pre-mix and the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

12. The feed supplement mix of claim 7, wherein the feed supplement mix comprises between about 60% and about 85% of the feed supplement pre-mix and the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

13. The feed supplement mix of claim 7, wherein the feed supplement mix comprises between about 70% and about 80% of the feed supplement pre-mix and the dewatered Asparagopsis spp. biomass comprises between about 60% and about 80% residual moisture.

14. The feed supplement mix of claim 7, wherein the feed supplement mix comprises between about 70% and about 80% of the feed supplement pre-mix and the dewatered Asparagopsis spp. biomass comprises between about 65% and about 75% residual moisture.

15. The feed supplement mix of claim 7, wherein the feed supplement mix comprises between about 70% and about 80% of the feed supplement pre-mix and the feed supplement pre-mix comprises between about 20% and about 40% edible oil.

16. The feed supplement mix of claim 7, wherein the feed supplement mix comprises between about 70% and about 80% of the feed supplement pre-mix and the feed supplement pre-mix comprises between about 25% and about 35% edible oil.

17. The feed supplement mix of claim 7, wherein the feed supplement mix comprises between about 60% and about 85% of the feed supplement pre-mix and the feed supplement pre-mix comprises between about 20% and about 40% edible oil.

18. The feed supplement mix of claim 7, wherein the feed supplement mix comprises between about 60% and about 85% of the feed supplement pre-mix and the feed supplement pre-mix comprises between about 25% and about 35% edible oil.

19. A feed supplement shot comprising: a) between about 40% and about 60% Asparagopsis spp. biomass, b) between about 17.5% and about 27.5% edible oil, c) between about 20% and about 30% carrier, d) between about 0.5% and about 3% xanthan gum, and e) between about 0.1% and about 2% palatability enhancer, wherein the feed supplement shot is formed by extrusion or pressing, wherein the Asparagopsis spp. biomass has a water content of between about 70% and about 80%; and, wherein the feed supplement shot has a moisture content of between about 30% and about 45%.

20. A feed supplement shot of claim 19 comprising: a) between about 65% and about 75% Asparagopsis spp. biomass, b) between about 20% and about 30% edible oil, c) between about 10% and about 20% carrier, c) between about 0.5% and about 3% xanthan gum, and d) between about 0.1% and about 2% palatability enhancer,wherein the feed supplement shot is formed by extrusion or pressing, wherein the Asparagopsis spp. biomass has a water content of between about 70% and about%; and, wherein the feed supplement shot has a moisture content of between about 40% and about%.

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