Methods for supporting grain intensive and / or energy intensive diets in ruminants with a synthetic bioensemble of microbes

Abstract

20 26 20 47 68 19 J un 2 02 6 A B S T R A C T 2 0 2 6 2 0 4 7 6 8 1 9 J u n 2 0 2 6

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 62 / 491,845, filed April 28, 2017; and U.S. Provisional Application No. 62 / 578,188, filed October 27, 2017; each of which is herein incorporated by reference in its entirety. FIELD

[0002] The present disclosure relates to isolated and biologically pure microorganisms that have applications, inter alia, in the farming of beef cattle. The disclosed microorganisms can be utilized m their isolated and biologically pure states, as well as being formulated into compositions. Furthermore, the disclosure provides a microbial ensemble, containing at least two members of the disclosed microorganisms, as well as methods of utilizing said microbial ensemble. Furthermore, the disclosure provides for methods of modulating the rumen microbiome. STATEMENT REGARDING SEQUENCE LISTING

[0003] The sequence li sting associated with this application is provided in text format in lieu of a paper copy, and is hereby incorporated by reference into the specification. The name of the text file containing the sequence listing is ASBI_005_02WO_ST25.txt. The text file is -4,544 kb, was created on April 27, 2018, and is being submitted electronically via EFS-Web. BACKGROUND

[0004] The global population is predicted to increase to over 9 billion people by the year 2050 with a concurrent reduction in the quantity of land, water, and other natural resources available 2026204768   19 Jun 2026 per capita. Projections indicate that the average domestic income will also increase, with the projected rise in the GDP of China and India. The desire for a diet richer in animal-source proteins rises in tandem with increasing income, thus the global livestock sector wall be charged with the challenge of producing more animal products using fewer resources. The Food and Agriculture Organization of the United Nations predict that 70% more food will have to be produced, yet the area of arable land available will decrease. It is clear that the food output per unit of resource input will have to increase considerably in order to support the rise in population.

[0005] Over recent decades the farm industry' has seen fast growth in the meat sector. As more of the world’s population ascends into the middle class demand for protein -- including beef -- is expected to remain robust for years to come. Worldwide beef production tops 59 million tons per annum.

[0006] Beef and products thereof are predominantly utilized in the preparation of foodstuffs in many different forms. There have been many strategies to improve beef production through nutritional modulations, hormone treatments, changes in animal management, and selective breeding; however, the need for more efficient production of edible beef foodstuffs per animal is required. Current animal feeding and handling practices, for example, often induce microbial dysbiosis in the rumen that ultimately leads to incidences of sub-acute acidosis or bloat, hindering the efficiency of production, increasing feed costs, and / or increasing a reliance on chemistry based treatments, such as antibiotics. 100071 Identifying compositions and methods for sustainably increasing beef production while balancing animal health and wellbeing have become imperative to satisfy the needs of everyday humans in an expanding population. Increasing the worldwide production of beef by scaling up the total number of beef cattle on farms would not only be economically infeasible for many parts of the world, but would further result in negative environmental consequences as the beef sector’s growth and trends towards intensification and concentration have already given rise to a number of environmental concerns, led predominantly by the production of far more waste than can be managed by land disposal. [0008| Population densities of beef cattle, particularly feedlot cattle, on large farms are often accompanied by an increased incidence of microbial pathogens that place the beef yield at risk, 2026204768   19 Jun 2026 and further place the ultimate consumer of the beef at risk in instances of zoonotic pathogens and / or the blooming of organisms in the rumen that lead to incidences of subacute acidosis (ruminal subacute acidosis) or bloat, which further hinders the productivity of feedlot operations. Considering the widespread occurrence of many zoonotic pathogens, it is unlikely that beef can be completely protected from exposure. Research has focused on investigative means of increasing resistance to colonization in beef cattle exposed to these pathogens.

[0009] Thus, meeting global beef yield expectations, by simply scaling up current high-input agricultural systems—utilized in most of the developed world—is simply not feasible.

[0010] There is therefore an urgent need in the art for improved methods of increasing beef production, while also mitigating the colonization and spread of microbial pathogens and further increasing the desirable aspects of beef. SUMMARY OF THE DISCLOSURE

[0011] In some aspects, the present disclosure provides isolated microbes, including novel strains of microbes, presented in Table 1 and / or Table 2.

[0012] In other aspects, the present disclosure provides isolated whole microbial cultures of the microbes identified in Table 1 and Table 2. These cultures may comprise microbes at various concentrations.

[0013] In some aspects, the disclosure provides for utilizing one or more microbes selected from Table 1 and / or Table 2 to increase a phenotypic trait of interest in beef cattle.

[0014] In some embodiments, a microbial composition comprises at least two microbial strains selected from Table 1 and / or Table 2. In another embodiment, a microbial composition is provided, said composition comprising at least one microbial strain selected from Table 1 and / or Table 2. In a further embodiment, a microbial composition comprises at least two microbial strains, wherein the at least two microbial strains comprise a 16S rRNA sequence encoded by sequences selected from SEQ ID NOs: 1-5993

[0015] In some embodiments, the disclosure is drawn to a ruminant supplement capable of treating or preventing acidosis or bloat in a ruminant, comprising: a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at 2026204768   19 Jun 2026 least about 97% identical to any one of SEQ ID NO: 1-5993: and b) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to treat or prevent acidosis or bloat in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0016] In some embodiments, the ruminant supplement capable of treating or preventing acidosis or bloat in a ruminant, comprising: a) a purified population of bacteria selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (li) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to treat or prevent acidosis or bloat in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0017] On some embodiments, the at least one of the bacteria are selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13

[0018] In one embodiment the purified population of bacteria is selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:75,

[0110] (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 13. |0111] In one embodiment, purified population of bacteria is selected from: (i) Succinivibrio bacteria wath a 16S nucleic acid sequence comprising SEQ ID NO:75, (ii) Prevotella bacteria wath a 16S nucleic acid sequence comprising SEQ ID NO:86, and / or (hi) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 13. 2026204768   19 Jun 2026

[0112] In one embodiment, the purified population of bacteria is selected from: (i) Succinivibrio bacteria deposited as B-67550. (ii) Prevotella bacteria deposited as B-67552, and / or (hi) Bacteroides bacteria deposited as B-67555.

[0113] In one embodiment the purified population of bacteria comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0114] In one embodiment, the ruminant supplement further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0115] In one embodiment, the ruminant supplement further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0116] In one embodiment the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, plastic, glass, polysaccharide, lipid, wax, oil, fatty acid, or glyceride.

[0117] In one embodiment, the encapsulated bacteria are vitrified. In one embodiment, the encapsulated bacteria are further encapsulated in a wax. In one embodiment, the purified population of bacteria are in the form of spores. In one embodiment, the spores are spray dried. In one embodiment, the ruminant supplement is formulated as a tablet, capsule, pill, feed additive, food ingredient, food additive, food preparation, food supplement, consumable solution, consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof. [Oil8| In some embodiments, the disclosure is drawn to a method for treating or preventing acidosis or bloat in a ruminant, comprising: administering to a ruminant an effective amount of a ruminant supplement comprising: a) a purified population of bacteria selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the 2026204768   19 Jun 2026 supplement in an amount effective to treat or prevent acidosis or bloat in a ruminant administered the supplement, as compared to a ruminant not administered the supplement. |0119] In some embodiments, at least one of the bacteria are selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

[0120] In some embodiments, the purified population of bacteria is selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 13.

[0121] In some embodiments, the purified population of bacteria is selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence comprising SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 13.

[0122] In some embodiments, the purified population of bacteria is selected from: (i) Succinivibrio bacteria deposited as B-67550. (ii) Prevotella. bacteria deposited as B-67552, and / or (hi) Bacteroides bacteria deposited as B-67555.

[0123] In some embodiments, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0124] In some embodiments, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0125] In some embodiments, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0126] In some embodiments, the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, sugar alcohol, surfactant, plastic, glass, polysaccharide, lipid, wax, oil, fatty acid, ammo acid, or glyceride. 2026204768   19 Jun 2026

[0127] In one embodiment, the encapsulated bacteria are vitrified. In one embodiment, the encapsulated bacteria are further encapsulated in a wax, fat, fatty acid, fatty alcohol, or glyceride. In one embodiment, the purified population of bacteria are in the form of spores. In one embodiment, the spores are spray dried. In one embodiment, the supplement is formulated as a tablet, capsule, pill, feed additive, food ingredient, food additive, food preparation, food supplement, consumable solution, consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof.

[0128] In one embodiment, the ruminant is administered to a ruminant orally. In one embodiment, the ruminant is a cow or a steer. In one embodiment, the ruminant is fed a step-up diet. In one embodiment, the ruminant is fed a finishing diet.

[0129] The ruminant supplement of claim 36, wherein the purified population of bacteria is selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 13.

[0130] In some embodiments, the purified population of bacteria is selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence comprising SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 13.

[0131] In some embodiments, the purified population of bacteria is selected from (i) Succinivibrio bacteria deposited as B-67550, (ii) Prevotella bacteria deposited as B-67552, and / or (iii) Bacteroides bacteria deposited as B-67555.

[0132] In some embodiments, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0133] In some embodiments, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0134] In some embodiments, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993. 2026204768   19 Jun 2026 10135] In some embodiments, the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, sugar alcohol, surfactant, plastic, glass, polysaccharide, lipid, wax, oil, fatty acid, amino acid, or glyceride. In some embodiment, the encapsulated bacteria are vitrified. In one embodiment, the encapsulated bacteria are further encapsulated in a wax, fat, fatty acid, fatty alcohol, or glyceride.

[0136] In one embodiment, the purified population of bacteria are in the form of spores. In one embodiment, the spores are spray dried. In one embodiment, the ruminant supplement is formulated as a tablet, capsule, pill, feed additive, food ingredient, food additive, food preparation, food supplement, consumable solution, consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof.

[0137] In some embodiments, the disclosure is drawn to a method of decreasing the amount of carbon dioxide and / or carbonic acid in the rumen of a ruminant, comprising: administering to a ruminant an effective amount of a ruminant supplement comprising: a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to decrease the amount of carbon dioxide and / or carbonic acid in the rumen of a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0138] In some embodiments, the disclosure is drawn to a method of decreasing the amount of carbon dioxide and / or carbonic acid in the rumen of a ruminant, comprising: administering to a ruminant an effective amount of a ruminant supplement comprising: a) a purified population of bacteria selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 86, and / or (Hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to decrease the amount of carbon dioxide and / or carbonic acid in the rumen of a ruminant administered the supplement, as compared to a ruminant not administered the supplement. 2026204768   19 Jun 2026

[0139] In one embodiment, at least one of the bacteria are selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

[0140] In one embodiment, thepurified population of bacteria is selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 13.

[0141] In one embodiment, the purified population of bacteria is selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence comprising SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 13.

[0142] In one embodiment, the purified population of bacteria is selected from : (i) Succinivibrio bacteria deposited as B-67550. (ii) Prevotella bacteria deposited as B-67552, and / or (iii) Bacteroides bacteria deposited as B-67555.

[0143] In one embodiment, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993

[0144] In one embodiment, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0145] In one embodiment, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0146] In one embodiment, the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, sugar alcohol, surfactant, plastic, glass, polysaccharide, lipid, wax, oil, fatty acid, ammo acid, or glyceride.

[0147] In one embodiment, the encapsulated bacteria are vitrified. In one embodiment, the encapsulated bacteria are further encapsulated in a wax, fat, fatty acid, fatty alcohol, or 2026204768   19 Jun 2026 glyceride.The method of claim 51, wherein the purified population of bacteria are in the form of spores. The method of claim 62, wherein the spores are spray dried. 10148] In one embodiment, the ruminant supplement is formulated as a tablet, capsule, pill, feed additive, food ingredient, food additive, food preparation, food supplement, consumable solution, consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof.

[0149] In one embodiment, the ruminant is administered to a ruminant orally. In one embodiment, the ruminant is a cow or a steer. In one embodiment, the ruminant is fed a step-up diet. In one embodiment, the ruminant is fed a finishing diet.

[0150] In one embodiment, the disclosure is drawn to a ruminant supplement capable of increasing the amount of meat marbling in a ruminant, comprising: a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993: and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase the amount of meat marbling in the ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0151] In one embodiment, the disclosure is drawn to a ruminant supplement capable of increasing the amount of meat marbling in a ruminant, comprising: a) a purified population of bacteria selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (li) Prevolella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (lii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase the amount of meat marbling in the ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0152] In one embodiment, the least one of the bacteria are selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (li) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13. 2026204768   19 Jun 2026 10153] In one embodiment, the purified population of bacteria is selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:86, and / or (hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 13.

[0154] In one embodiment, the purified population of bacteria is selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence comprising SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 13.

[0155] In one embodiment, the purified population of bacteria is selected from :(i) Succinivibrio bacteria deposited as B-67550, (ii) Prevotella bacteria deposited as B-67552, and / or (iii) Bacteroides bacteria deposited as B-67555.

[0156] In one embodiment, the rumen supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0157] In one embodiment, the rumen supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993. [015§] In one embodiment, the rumen supplement further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0159] In one embodiment, the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, sugar alcohol, surfactant, plastic, glass, polysaccharide, lipid, wax, oil, fatty acid, amino acid, or glyceride.

[0160] In one embodiment, the encapsulated bacteria are vitrified. In one embodiment, the encapsulated bacteria are further encapsulated in a wax, fat, fatty acid, fatty alcohol, or glyceride. In one embodiment, the purified population of bacteria are in the form of spores. In one embodiment, the spores are spray dried.

[0161] In one embodiment, the rumen supplement is formulated as a tablet, capsule, pill, feed additive, food ingredient, food additive, food preparation, food supplement, consumable solution, 2026204768   19 Jun 2026 consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof. 10162] In some embodiments, the disclosure is drawn to a method increasing the amount of meat marbling in a ruminant, comprising: administering to a ruminant an effective amount of a ruminant supplement comprising: a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase the amount of meat marbling in the ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0163] In some embodiments, the disclosure is drawn to a method increasing the amount of meat marbling in a ruminant, comprising: administering to a ruminant an effective amount of a ruminant supplement comprising: a) a purified population of bacteria selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; and b) a earner suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase the amount of meat marbling in the ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0164] In one embodiment, at least one of the bacteria are selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about

[0165] 97% identical to SEQ ID NO: 13.

[0166] In one embodiment, the purified population of bacteria is selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 2026204768   19 Jun 2026

[0167] 99% identical to SEQ ID NO: 13.

[0168] In one embodiment, the purified population of bacteria is selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence comprising SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 86, and / or (in) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 13.

[0169] In one embodiment, the purified population of bacteria is selected from : (i) Succinivibrio bacteria deposited as B-67550, (ii) Prevotella bacteria deposited as B-67552, and / or (iii) Bacteroides bacteria deposited as B-67555.

[0170] In one embodiment, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO:5993.

[0171] In one embodiment, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO:5993.

[0172] In one embodiment, the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

[0173] In one embodiment, the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, sugar alcohol, surfactant, plastic, glass, polysaccharide, lipid, wax, oil, fatty acid, amino acid, or glyceride.

[0174] In one embodiment, the encapsulated bacteria are vitrified. In one embodiment, the encapsulated bacteria are further encapsulated in a wax. On one embodiment, the purified population of bacteria are in the form of spores. In one embodiment, the spores are spray dried.

[0175] In one embodiment, the ruminant supplement is formulated as a tablet, capsule, pill, feed additive, food ingredient, food additive, food preparation, food supplement, consumable solution, consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof. In one embodiment, the ruminant supplement is administered to a ruminant orally. In one embodiment, the ruminant is a cow or a steer. In one embodiment, the ruminant is fed a step-up diet. In one embodiment the ruminant is fed a finishing diet. In one embodiment, the increase in meat marbling is an increase of at least 10%. 2026204768   19 Jun 2026

[0176] In some embodiments, the disclosure is drawn to a ruminant supplement capable of increasing feed efficiency in a ruminant, comprising: a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993: and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase feed efficiency in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0177] In some embodiments, the disclosure is drawn to a ruminant supplement capable of increasing feed efficiency in a ruminant, comprising: a) a purified population of bacteria selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase feed efficiency in a ruminant administered the supplement's compared to a ruminant not administered the supplement.

[0178] In some embodiments, the at least one of the bacteria are selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

[0179] In some embodiments, the disclosure is drawn to a method for increasing feed efficiency in a ruminant, comprising: administering to a ruminant an effective amount of a ruminant supplement comprising: a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase feed efficiency in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0180] In some embodiments, the method increasing feed efficiency in a ruminant comprises: administering to a ruminant an effective amount of a ruminant supplement comprising: a) a 2026204768   19 Jun 2026 purified population of bacteria selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (li) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 86, and / or (in) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase feed efficiency in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0181] In some embodiments, the at least one of the bacteria are selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

[0182] In some embodiments, the ruminant supplement capable of increasing performance in a ruminant, comprising: a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993: and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase performance in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0183] In some embodiments, the ruminant supplement capable of increasing performance in a ruminant, comprising: a) a purified population of bacteria selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase performance in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0184] In some embodiments, at least one of the bacteria are selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, 2026204768   19 Jun 2026 (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

[0185] In some embodiments, the disclosure is drawn to a ruminant supplement capable of reducing methane production and emission in a ruminant, comprising: a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993: and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to reduce methane production and emission in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0186] In some embodiments, the disclosure is drawn to a ruminant supplement capable of reducing methane production and emission in a ruminant, comprising: a) a purified population of bacteria selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 86, and / or (hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in anamount effective to reduce methane production and emission in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0187] In some embodiments, at least one of the bacteria are selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or (hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

[0188] In some embodiments, the disclosure is drawn to a ruminant supplement capable of reducing methane production and emission in a ruminant, comprising: a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993: and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the 2026204768   19 Jun 2026 supplement in an amount effective to reduce methane production and emission in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

[0189] In some embodiments, the disclosure is drawn to a ruminant supplement capable of reducing methane production and emission in a ruminant, comprising: a) a purified population of bacteria selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 86, and / or (hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; and b) a carrier suitable for ruminant administration; wherein the purified population of bacteria of a) is present in the supplement in an amount effective to reduce methane production and emission in a ruminant administered the supplement,as compared to a ruminant not administered the supplement.

[0190] In some embodiments, the at least one of the bacteria are selected from: (i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75, (ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 86, and / or (iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

[0191] In some embodiments, the microbes are administered with a prebiotic, a vitamin, or a mineral. In some embodiments, the microbes are administered with vitamin B or a precursor thereof. BUDAPEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR THE PURPOSE OF PATENT PROCEDURES

[00192] The microorganisms described in this Application were deposited with (1) the American Type Culture Collection (ATCC4), located at 10801 University Blvd., Manassas, VA 20110, USA; and the United States Department of Agriculture (USDA) Agricultural Research Service (ARS) Culture Collection (NRRl. ). located at 1815 N. University St., Peoria, IL 61604, USA

[00193] The deposits were made under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. The ATCC and NRRL accession numbers for the aforementioned Budapest Treaty 2026204768   19 Jun 2026 deposits are provided in Table 1. The Accession numbers and corresponding dates of deposit for the microorganisms described in this Application are separately provided in Table 2.

[00194] The strains designated in the below table have been deposited in the labs of Ascus Biosciences, Inc. since at least April 22, 2017, and August 2017.

[00195] In Table 1, the closest predicted hits for taxonomy of the microbes are listed in columns 2, and 5. Column 2 is the top taxonomic hit predicted by BLAST, and column 5 is the top taxonomic hit for genus + species predicted by BLAST. The strains designated in the below table have been deposited in the labs of Ascus Biosciences, Inc. since at least April 22, 2017, and August,                                                                             2017. 2026204768   19 Jun 2026 Table 1: Microbes of the present disclosure, including bacteria (1-190). Predicted Closest Taxa of Isolated Microbes BLAST Taxonomic Top Hit w / Genus + Species BLAST Ident. Query Cover Strain Designation Sequence Identifier for Associated Marker MIC Score 1. Prevotella (genus) Prevotella rumimcola 93% 98% Ascusbbf 6176 SEQID NO: I 1 2. Prevotella (genus) Prevotella loescheii 88% 99% Ascusbbf 22143 SEQID NO :2 1 3. Prevotella (genus) Prevotella rumimcola 91% 100% Ascusbbf 4883 SEQID NO :3 0.97095 4. Selenomonas (genus) Selenomonas ruminantium 93% 95% Ascusbbf 13543 SEQID NO :4 0.97095 5. Clostridium XlVa (Cluster) Oscillibacter valericigenes 92% 100% Ascusbbf 152 SEQID NO :5 0.88129 6. Clostridium XlVa (Cluster) Oscillibacter valericigenes Ascusbbf 152A SEQID NO:5398 0.88129 7. Prevotella (genus) Prevotella ruminicola 94% 100% Ascusbbf 707 SEQID NO :6 0.88129 8. Fibrobacter (genus) Fibrobacter intestinalis 99% 100% Ascusbbf 1238 SEQID NO :7 0.88129 9. Prevotella (genus) Prevotella rumimcola 89% 100% Ascusbbf 5588 SEQID NO :8 0.88129 10. Saccharofermentans (genus) Saccharofermentans acetigenes 86% 100% Ascusbbf 4691 SEQID NO :9 0.88129 11. Saccharofermentans (genus) Saccharofermentans acetigenes Ascusbbf 469IC SEQID NO:5425 0.88129 12. Saccharofermentans (genus) Intestinimonas b utyri ci producens 86% 100% Ascusbbf 59499 SEQID NOTO 0.88129 13. Bacillus (genus) Brevibacillus brevis 86% 89% Ascusbbf 9770 SEQID NO: 11 0.88129 2026204768   19 Jun 2026 14. Spirochaeta (genus) Treponema parvum | 88% 92% I SEQ ID Ascusbbf 123632 NO: 12 0.88129 15. Bacleraides (genus) Baderoides     | xylanisolvens     | 99% 97% SEQ ID Ascusbbf 14146 | NO:13 0.78606 16. Lachnospiracea incertae seais (genus) Desulfotomacuhim sp. | 94% 100% I SEQ ID Ascusbbf 1103      NO: 14 0.67032 17. Clostridium XlVa (Cluster) Lachnoclostridium i pacaense       1 89% 100% i SEQ ID Ascusbbf 498      NO: 15 0.66823 18. Prevotella (genus) Prevotella oralis 1 89% 100% i SEQ ID Ascusbbf 13717     NO: 16 0.65415 19. Prevotella (genus) Prevotella oralis 1 i SEQ ID Ascusbbf 13717A   NO:5450 0.65415 20. Clostridium XlVa (Cluster) Coprococcus catus 1 90% 97% SEQ ID Ascusbbf 876   | NO: 17 0.65106 21. Bacleroides (genus) Bacteroides unifonnis 1 89% 100% i SEQ ID Ascusbbf 612      NO: 18 0.65002 22. Selenomonas (genus) Selenomonas     | niminantium     1 97% 100% i SEQ ID Ascusbbf 4936      NO: 19 0.63816 23. Selenomonas (genus) Selenomonas     | niminaiitium      1 SEQ ID Ascusbbf 4936A | NO:5460 0.63816 24. Prevotella (genus) Prevotella oulorum 1 92% 100% i SEQ ID Ascusbbf 6809     NO:20 0.6337 25. Clostridium XlVa (Cluster) Clostridium      | aminophilum    1 91% 100% i SEQ ID Ascusbbf 113152 | NO:21 0.63008 26. Clostridium XlVa (Cluster) Clostridium      | aminophilum     1 i SEQ ID Ascusbbf 113152A  NO:5462 0.63008 27. Ruminococcus (genus) Ruminococcus bromii 1 99% 96% SEQ ID Ascusbbf 18    1 NO:22 0.62713 28. Prevotella (genus) Prevotella mminicola I 94% 100% i SEQ ID Ascusbbf 9031   1 NO:23 0.62075 29. Spirochaeta (genus) Treponema      | brennaborense    | 86% 98% i SEQ ID Ascusbbf 11823     NO:24 0.61287 2026204768   19 Jun 2026 30. Butyricimonas (genus) Porphyromonadaceae 1 87% 98% I SEQ ID Ascusbbf 1007      NO:25 0.60495 31. Butyricimonas (genus) Porphyromonadaceae 1 SEQ ID Ascusbbf 1007A | NO:5473 0.60495 32. Prevotella (genus) Prevotella baroniae 1 87% 99% SEQ ID Ascusbbf 24422 | NO:26 0.59156 33. Prevotella (genus) Prevotella baroniae 1 i SEQ ID Ascusbbf 24422A   NO:5474 0.59156 34. OlsenePia (genus) Olsenella umbonata 1 99% 100% SEQ ID Ascusbbf 951   | NO:27 0.59007 35. Clostridium XlVa (Cluster) [Clostridium]      | symbiosum     1 96% 100% i SEQ ID Ascusbbf 80169     NO:28 0.58852 36. Spirochaeta (genus) Treponema biyantii 1 90% 97% SEQ ID Ascusbbf 5699   | NO:29 0.58423 37. Spirochaeta (genus) Treponema biyantii 1 i SEQ ID Ascusbbf 5699B    NO:5483 0.58423 38. Prevotella (genus) Prevotella ruminicola | 91% 100% i SEQ ID Ascusbbf 130      NO:30 0.58333 39. Acidaminococcus (genus) Acidaminococcus | fermentans      1 95% 100% i SEQ ID Ascusbbf 10109     NO:31 0.58267 40. Parabacteroides (genus) Culturomica      | massiliensis      1 86% 89% i SEQ ID Ascusbbf 29797     NO:32 0.58241 41. Parabacteroides (genus) Culturomica      | massiliensis       1 i SEQ ID Ascusbbf 29797A   NO:5502 0.58241 42. Clostridium sensu stricto (genus) Cliristensenella     I limonensis      1 86% 99% i SEQ ID Ascusbbf 24410     NO:33 0.58142 43. Oribacterium (genus) Oribacterium sinus 1 91% 99% i SEQ ID Ascusbbf 54068 1 NO:34 0.58113 44. Clostridium XI Va (Cluster) [Clostridium] bolteae 1 93% 100% i SEQ ID Ascusbbf 7003   1   NO:3.5 0.58059 4 5. Pseudoflavonifractor (genus) Intestinimonas | butyticiproducens I 89% 100% i SEQ ID Ascusbbf 23       NO:36 0.57785 2026204768   19 Jun 2026 46. Prevoiella (genus) Prevoiella ruminicola 1 90% 100% I SEQ ID Ascusbbf 1697   1 NO:37 0.57337 47. Prevoiella (genus) Prevoiella ruminicola 1 i SEQ ID Ascusbbf 1697B    NO:5511 0.57337 48. Treponema (genus) Treponema zioleckii | 99% 99% I SEQ ID Ascusbbf 24513     NO:38 0.5696 49. Prevoiella (genus) Prevoiella oralis       89% 100% i SEQ ID Ascusbbf 7586     NO:39 0.56896 50. Butyricimonas (genus) Bamesiella viscericola | 85% 91% i SEQ ID Ascusbbf 27854     NO:40 0.56657 51. Saccharofermentans (genus) Oscillibacter      | valericigenes     1 87% 100% i SEQ ID Ascusbbf 1034      NO:41 0.56476 52. Saccharofermentans (genus) Oscillibacter      | valericigenes      | SEQ ID Ascusbbf 1034A | NO:5517 0.56476 53. Butyricimonas (genus) Butyricimonas virosa 1 82% 100% i SEQ ID Ascusbbf 23134     NO:42 0.56219 54. Butyricimonas (genus) Butyricimonas virosa 1 i SEQ ID Ascusbbf 23134A   NO:5519 0.56219 55. Rhodobacter (genus) Gemmobacter    | intermedins      1 99% 87% SEQ ID Ascusbbf 7027   | NO:43 0.56127 56. Prevoiella (genus) Butyricimonas virosa 1 84% 92% i SEQ ID Ascusbbf 43679     NO:44 0.56056 57. Fluviicola. (genus) Anaerocella delicate 85% 87% i SEQ ID Ascusbbf 63954     NO:45 0.55952 58. Fluviicola (genus) Anaerocella delicate 1 SEQ ID Ascusbbf 63954A | NO:5526 0.55952 59. Succiniclasticum (genus) Succiniclasticum | ruminis       1 95% 95% i SEQ ID Ascusbbf 1517   | NO:46 0.55908 60. Solobacterium (genus) Solobacterium moorei 1 91% 99% i SEQ ID Ascusbbf 104   1 NO:47 0.55759 61. Clostridium XlVa (Cluster) [Clostridium]      | lavalense       | 90% 100% i SEQ ID Ascusbbf 148      NO:48 0.55551 2026204768   19 Jun 2026 62. Brevotella (genus) Prevotella bryantii 1 99% 100% Ascusbbf 944 I SEQ ID 1 NO:49 0.55265 63. Lachnospiracea incertae Eubacterium     । i SEQ ID seals (.genus) oxidoreducens 1 90% 100% Ascusbbf 76009 NO:50 0.55253 64. Veillonella (genus) Holdemania filiformis 1 84% 96% Ascusbbf 23033 I SEQ ID NO:51 0.55253 65. Cellulosimicrobium Cellulosimicrobium । i SEQ ID (genus) cellulans         1 95% 100% Ascusbbf 20389 NO:52 0.55131 66. Cupriavidus (genus) Sutterella         | wads worthensis    1 92% 100% Ascusbbf 2600 i SEQ ID NO:53 0.54892 67. Bacteroides (genus) Paraprevotella     | xylaniphila       | 86% 92% Ascusbbf 8118 i SEQ ID NO:54 0.54888 68. Brevotella (genus) Prevotella ruminicola | 92% 100% Ascusbbf 201 i SEQ ID NO:55 0.54656 69. Brevotella (genus) Prevotella ruminicola | Ascusbbf 20IK i SEQ ID NO:5576 0.54656 70. Spirochaeta (genus) Treponema      | saccharophilum    | 88% 100% Ascusbbf 6315 i SEQ ID NO:56 0.54535 71. Megasphaera (gemis) Megasphaera elsdenii | 99% 100% Ascusbbf 10712 i SEQ ID NO:57 0.54494 72. Megasphaera (genus) Megasphaera elsdenii | Ascusbbf 10712E i SEQ ID NO:5582 0.54494 73. Succinivibrio (genus) Succinivibrio      | dextrinosolvens    | 90% 99% Ascusbbf 6012 i SEQ ID 1 NO:58 0.54428 74. Succinivibrio (genus) Succinivibrio      | dextrinosolvens    | Ascusbbf 60I2C i SEQ ID NO:5589 0.54428 75. Spirochaeta (genus) Treponema bryantii 1 98% 99% Ascusbbf 2297 i SEQ ID 1 NO:59 0.54413 76. Spirochaeta (genus) Treponema bryantii 1 Ascusbbf 2297G i SEQ ID 1 NO:5598 0.54413 77 Bacteroides (genus) Bacteroides uniformis | 89% 100% Ascusbbf 9540 i SEQ ID NO:60 0.54383 2026204768   19 Jun 2026 78. Oscillibacter (genus) Oscillibacter      | valericigenes      | 94% 100% Ascusbbf 873    1 SEQID NO:61 0.54374 79. Prevotella (genus) Prevotella dentalis 1 83% 95% Ascusbbf 87102 | SEQID NO:62 0.54356 80. Pseudomonas (genus) Pseudomonas     | pertucinogena     | 98% 99% Ascusbbf 77105 | SEQID NO:63 0.54356 81. Corynebacterium (genus) Corynebacterium | marinum       | 99% 100% Ascusbbf 269   | SEQID NO:64 0.54206 82. Adlercreutzia (genus) Raoultibacter     | massiliensis       | 92% 100% Ascusbbf 41015 1 SEQID NO:65 0.54192 83. Adlercreutzia (genus) Raoultibacter     | massiliensis       | Ascusbbf 41015A | SEQID NO:5614 0.54192 84. Acidaminococcus (genus) Acidaminococcus | fermentans       | 98% 97% Ascusbbf 32877 1 SEQID NO:66 0.54166 85. Acidaminococcus (genus) Acidaminococcus | fermentans       | Ascusbbf 32877A | SEQID NO:5619 0.54166 86. Dorea (genus) Dorea longicatena | 99% 100% Ascusbbf 57294 | SEQID NO:67 0.53443 87. Dorea (genus) Dorea longicatena | Ascusbbf 57294B | SEQID NO:5621 0.53443 88. Roseburia (genus) Howardella ureilvtica 1 88% 98% Ascusbbf 27932 | SEQID NO:68 0.53375 89. Anaerovibrio (genus) Anaerovibrio      | lipolyticus        | 95% 97% Ascusbbf 22558 | SEQID NO:69 0.53353 90. Anaerovibrio (genus) Anaerovibrio      | lipolyticus        | Ascusbbf 22558B 1 SEQID NO:5627 0.53353 91. Bacteroides (genus) Bacteroides      | helcogenes       1 88% 100% Ascusbbt 983757 | SEQID NO:70 0.5317 92. Bacteroides (genus) Bacteroides      | helcogenes       1 Ascusbbf 983757B I SEQID NO:5629 0.5317 93. Clostridium XlVa (Cluster) Clostridium      | aminophilum     | 98% 100% Ascusbbf 52330 1 SEQID NO:71 0.53133 2026204768   19 Jun 2026 94. Clostridium XI Va (Cluster) Clostridium      | aminophilum     1 Ascusbbf 52330A I SEQ ID 1 NO:5631 0.53133 95. Sporosarcina (genus) 96. Slreptomyces (genus) Lactobacillus floricola 1 -----------------------------------------------+-- Streptomyces albus 1 79% 99% 97% 100% Ascusbbf_88445 Ascusbbf 4111 i SEQ ID NO:72 I SEQ ID NO:73 0.53069 0.53006 97. Syntrophococcus (genus) Syntrophococcus   | sucromutans      1 93% 100% Ascusbbf 1085 i SEQ ID NO:74 0.5294 98. Succinivibrio (genus) Succinivibrio      | dextrinosolvens    1 99% 99% Ascusbbf 154 i SEQ ID NO:75 0.52737 99. Selenomonas (genus) Selenomonas bovis 1 99% 100% Ascusbbf 1010 i SEQ ID NO:76 0.527 100. Parabacteroides (genus) Megasphaera indica | 99% 99% Ascusbbf 5575 i SEQ ID NO:77 0.52675 101. Parabacteroides (genus) Megasphaera indica | Ascusbbf 5575B i SEQ ID NO:5663 0.52675 102. Prevotella (genus) Prevotella oris | 82% 100% Ascusbbf 775 i SEQ ID 1 NO:78 0.52672 103. Prevotella (genus) Prevotella oris | Ascusbbf 775A i SEQ ID NO:5670 0.52672 104. Butyrivibrio (genus) Butyrivibrio       | fibrisolvens       1 96% 100% Ascusbbf 19348 i SEQ ID NO:79 0.52608 105. Clostridium sensu stricto (genus) Clostridium      I beijerinckii        | 99% 100% Ascusbbf 24302 i SEQ ID NO :80 0.52361 106. Succinivibrio (genus) Succinivibrio      | dextrinosolvens    1 99% 97% Ascusbbf 1 i SEQ ID NO:81 0.51924 107. Lachnobacterium (genus) Lachnobacterium | bovis          1 99% 99% Ascusbbf 52548 i SEQ ID 1 NO :82 0.51683 108. Clostridium IV (Cluster) Closlridiales bacterium 1 9.3% 100% Ascusbbf 50658 i SEQ ID 1 NO:83 0.51263 109. Lachnospiracea incertae sedis (genus) Lachnospira      | pectinoschiza     I 89% 100% Ascusbbf 850 i SEQ ID NO :84 0.5088 2026204768   19 Jun 2026 110. Parabacteroides (genus) Parabacteroides    | distasonis       1 84% 100% I SEQ ID Ascusbbf 25259 1 NO:85 0.50691 111,   Prevote Ila (genus) Prevotella albensis | 98% 100% SEQ ID Ascusbbf 4    | NO :86 0.50464 112, Bacteroides (genus) Bacteroides umfomris | 89% 100% I SEQ ID Ascusbbf 5131      NO :87 0.49238 113. Clostridium IV (Cluster) Caproiciproducens | galactitolivorans 1 89% 95% i SEQ ID Ascusbbf 8600     NO:88 0.47814 114. Clostridium IV (Cluster) Caproiciproducens | galactitolivorans 1 i SEQ ID Ascusbbf 8600B    NO:5726 0.47814 115. Pyramidobacter (genus) Rarimicrobium    I hominis       1 92% 94% i SEQ ID Ascusbbf 1273      NO:89 0.46972 116. Ruminococcus (genus) Ruminococcus    | flavefaciens      1 98% 99% i SEQ ID Ascusbbf 39159     NO:90 0.46727 117. Coprococcus (genus) Eubacterium     | oxidoreducens    1 89% 100% SEQ ID Ascusbbf 9751   | NO:91 0.4618 118. Ruminobacter (genus) Ruminobacter    | amylophilus        99% 100% i SEQ ID Ascusbbf 318      NO :92 0.45953 119. Thermobifida (genus) Thermobifida fusca 1 99% 100% SEQ ID Ascusbbf 7046   | NO:93 0.45752 120. Papillibacter (genus) Oscillibacter      | valericigenes     1 86% 100% i SEQ ID Ascusbbf 25993     NO:94 0.45023 121. Rhodobacter (genus) Rhodobacter     | gluconicum     1 95% 99% Ascusbbi o,    ।  N0.95 0.56127 122.   Prevote Ila (genus) Gabonibacter     | massiliensis      | 87% 76% ' u.r mines i SEQ ID Ascusbbf 13 72985 i .,2-.. ~         ।  NO:96 0.55953 123.   Prevote Ila (genus) Gabonibacter     | massiliensis       | 0.55953 124. Aquamarina atlantica (genus + species) Gabonibacter     | massiliensis      1 89% 76% 1 SEQ ID Ascusbbf 23253 1 NO:97 0.50683 125. Aquamarina pacifica (genus + species) Gabonibacter     | massiliensis      | 86% 87% 1 SEQ ID Ascusbbf 121971     NO:98 0.42 2026204768   19 Jun 2026 126. Aquamarina pacifica (genus + species) Gabonibacter     | massiliensis       1 Ascusbbf 121971A 1 SEQID NO:5757 0.42 127. Treponema bryantii (genus + species) Treponema bryantii 1 98% 94% Ascusbbf 5251   | SEQID NO:99 0.56738 128. Treponema bryantii (genus + species) T repone tna bryantii   | Ascusbbf 5251G | SEQID NO:5764 0.56738 129. Actinomyces turicensis (genus + species) Actinomyces turicensis 1 85% 100% Ascusbbf 6716   | SEQID NO: 100 0.5201 130. Prevolella (genus) Prevolella oulorum 1 92% 99% Ascusbbf 100    | SEQID NO: 101 0.53729 131.   Staphylococcus (genus) Paenibacillus     1 hemerocallicola    1 84% 84% Ascusbbf 20584 | SEQID NO: 102 0.52104 132 Prevolella (genus) Prevotella rumimcola | 88% 100% Ascusbbf 4317   | SEQID NO: 103 0.55564 133. Prevolella (genus) Prevotella rumimcola | Ascusbbf 4317D | SEQID NO:5777 0.55564 134 Prevolella (genus) Prevotella ruminicola 1 90% 93% Ascusbbf 6     | SEQID NO: 104 0.46763 135. Mogibacterium (genus) Mogibacterium    i pumilum       1 91% 100% Ascusbbf 19022 1 SEQID NO: 105 0.47803 13 6.   P sen dob u tyrib i brio (genus) Pseudobutyrivibrio I rumims        | 99% 100% Ascusbbf 2624   | SEQID NO: 106 0.52337 13 7.   P sen dob u tyrib i brio (genus) Pseudobutyrivibrio i rumims        | Ascusbbf 2624D | SEQID NO:5797 0.52337 138. Fluviicola (genus) Fluviicola taffensis | 84% 90% Ascusbbf 3427   | SEQID NO: 107 0.50515 139. Fluviicola (genus) Fluviicola tatfensis 1 Ascusbbf 3427B 1 SEQID NO:5802 0.50515 140.   Prevote Ila (genus) Prevotella rumimcola 1 92% 100% Ascusbbf 5005 SEQID NO: 108 0.57034 141.   Prevote Ila (genus) Prevotella rumimcola | 100% 100% Ascusbbf 69    | SEQID NO: 109 0.50536 2026204768   19 Jun 2026 142. (P Succiniclasticum enus) Succiniclasticum    | ruminis        1 90% 90% Ascusbbf 8082 I SEQ ID NO: 110 0.50084 143. Prevolella (genus) Prevotella rumimcola | 94% 100% Ascusbbf 95 i SEQ ID NO:111 0.53509 144. Clostridium XlVa (cluster) Butyrivibrio       । fibrisolvens       1 89% 100% Ascusbbf 1136 I SEQ ID NO: 112 0.50966 145. Asteroleplasma (genus) Asteroleplasma    | anaerobium      | 98% 100% Ascusbbf 2770 i SEQ ID NO: 113 0.51006 146 Turicibacter (genus) Turicibacter sanguinis | 98% 100% Ascusbbf 1629 i SEQ ID NO: 114 0.51632 147. Prevolella (genus) Bacteroides caecicola | 85% 99% Ascusbbf 1821 i SEQ ID NO: 115 0.53784 148. Prevolella (genus) Prevotella rumimcola 1 95% 100% Ascusbbf 56782, i SEQ ID NO: 116 0.5.317 149. Olsenella (genus) Olsenella scatoligenes | 99% 100% Ascusbbf 92 i SEQ ID NO: 117 0.46089 150. Prevotella (genus) Prevotella rumimcola | 94% 99% Ascusbbf 118 i SEQ ID NO: 118 0.6108 15 1. Prevolella (genus) Prevotella rumimcola | Ascusbbf 118B i SEQ ID NO:5868 0.6108 152. Aggregatibacter (genus) Prevotella rumimcola | 87% 44% Ascusbbf 5429 i SEQ ID NO: 119 0.57983 153. Aggregatibacter (genus) Prevotella rumimcola | Ascusbbf 5429C i SEQ ID NO:5872 0.57983 154. Ruminobacter (genus) Ruminobacter     i amylophilus      1 86% 88% Ascusbbf 3 i SEQ ID NO :120 0.56323 155. Prevotella (genus) Prevotella rumimcola 1 91% 99% Ascusbbf 10576 i SEQ ID 1 NO: 121 0.562.08 156. Prevotella (genus) Prevotella rumimcola 1 93% 98% Ascusbbf 729 i SEQ ID 1 NO: 122 0.54949 157. Prevotella (genus) Prevotella rumimcola | 92% 100% Ascusbbf 201 i SEQ ID NO: 123 0.54656 2026204768   19 Jun 2026 158. Prevotella (genus) Prevotella ruminicola 1 91% 99% I SEQ ID Ascusbbf 416   1 NO: 124 0.53816 159, Prevolella (genus) Prevotella rumimcola | 94% 99% SEQ ID Ascusbbf 15806 | NO: 125 0.52527 160. Clostridium XlCa (cluster) Clostridium      i aminophilum     1 94% 100% I SEQ ID A scusbbf 6115     NO :126 0.52278 161, Anaerovibrio (genus) Anaerovibrio     i lipolyticus       1 95% 97% i SEQ ID Ascusbbf 1325058 NO: 127 0.52183 162, Prevolella (genus) Prevotella buccalis | 91% 100% i SEQ ID Ascusbbf 28350    NO: 128 0.51744 163. Parabacteroides (genus) Muribaculum     । intestmale       1 93% 100% i SEQ ID Ascusbbf 372   | NO: 129 0.51572 164. Phascolarctobacterium (genus) Phascolarctobacterium i succinatutens     1 96% 93% SEQ ID Ascusbbf 667   | NO: 130 0.51381 16 5. Phascolarctobacteri um (genus) Phascolarctobacterium i succinatutens     1 SEQ ID Ascusbbf 667A | NO:5930 0.51381 166. Bacleroides (genus) Bacteroides      i coprophilus      1 83% 100% i SEQ ID Ascusbbf 1207     NO: 131 0.51075 167. Lachnospiracea incertae sedis (genus) Coprococcus catus | 92% 97% i SEQ ID Ascusbbf 3875     NO: 132 0.47237 168. Clostridium XlCa (cluster) Clostridium      i aminophilum     | 90% 100% i SEQ ID Ascusbbf 72889    NO: 133 0.46531 169. Clostridium XlCa (cluster) Clostridium      i aminophilum     1 i SEQ ID Ascusbbf 72889B   NO:5947 0.46531 170. Parabacteroides (genus) Barnesiella viscericola । 85% 94% i SEQ ID Ascusbbf 106863 NO: 134 0.45152 171. Parabacteroides (genus) Barnesiella viscericola i i SEQ ID Ascusbbf 106863B 1 NO:5949 0.45152 172.   Prevote Ila (genus) prevotella ruminicola 1 94% 97% i SEQ ID Ascusbbf 120   1 NO: 135 0.53376 173.   Prevote Ila (genus) prevotella ruminicola | 93% 99% SEQ ID Ascusbbf 930   | NO: 136 0.51321 2026204768   19 Jun 2026 174, Bacteroides (genus) Bacteroides uniformis 1 89% 100% Ascusbbf 915 I SEQ ID 1 NO :5369 0.54396 175, Bacteroides (genus) Bacteroides uniformis | Ascusbbf 915A SEQ ID 1 NO:5955 0.54396 176, Prevoteila (genus) Prevoteila ruminicola | 88% 98% Ascusbbf 8941 I SEQ ID NO:5370 0.5328 177, Prevoteila (genus) Prevoteila ruminicola | Ascusbbf 8941A i SEQ ID NO:5956 0.5328 178, Anaerovobrio Anaerovibrio     i lipolyticus         | 96% 97% Ascusbbf 8480 i SEQ ID NO:5371 0.48193 179, Anaerovobrio Anaerovibrio     | lipolyticus         1 Ascusbbf 8480A i SEQ ID NO:5989 0.48193 180, Prevoteila (genus) prevoteila ruminicola | 92% 98% Ascusbbf 374 i SEQ ID NO::5372 0.52368 181, Prevoteila (genus) prevoteila ruminicola | Ascusbbi 3 74C i SEQ ID NO:5991 0.52368 182, Prevoteila (genus) Prevoteila brevis 1 92% 100% Ascusbbf 6906 i SEQ ID NO::5373 0.4889 183, Prevoteila (genus) Prevoteila ruminicola 1 94% 99% Ascusbbf 721 i SEQ ID NO:5374 0.88129 184. Syntrophococcus (genus) Syntrophococcus   । sucromutans      | 93% 100% Ascusbbf 3819 i SEQ ID NO:5375 0.45798 185. Syntrophococcus (genus) Syntrophococcus   i sucromutans      | Ascusbbf 3819A i SEQ ID NO:5971 0.45798 186. Bacteroides (genus) Bacteroides coprocola | 87% 100% Ascusbbf 4323 i SEQ ID NO:5376 0.50634 187. Bacteroides (genus) Bacteroides coprocola 1 Ascusbbf 4323B i SEQ ID 1 NO:5973 0.50634 188. Prevoteila (genus) Prevoteila ruminicola 1 91% 100% Ascusbbf 6087 SEQ ID NO:5377 0.52954 189. Prevoteila (genus) Prevoteila ruminicola | Ascusbbf 6087B SEQ ID NO :5 977 0.52954 2026204768   19 Jun 2026 190. Saccharofermentans (genus) Christensenella timonensis 86% 99% Ascusbbf 8414 SEQ ID NO:5378 0.52719 Table 2: Deposited Microbes of the present disclosure Strain Designation SEQ ID No: Deposit Accession # Strain Designation SEQ ID No: Deposit Accession # Strain Designation SEQ ID No: Deposit Accession # Ascusbbf 6176A 5379 PTA-125041, PTA-125O42,PTA-125049,PTA-125050, PTA-125051,PTA-125052 Ascusbbf 10712F 5583 PTA- 125033, PTA- 125042, PTA- 125050 Ascusbbf 6E 5783 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125052 Ascusbbf 6176B 5380 PTA- 125041 ,PTA- 125042 ,PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 10712G 5584 PTA- 125033, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125052 Ascusbbf 6F 5784 PTA-12.5040, PTA-125052 Ascusbbf 6176C 5381 PTA- 125041 ,PTA- 125049.PTA- 125050 Ascusbbf 10712H 5585 PTA- 125033, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125052 Ascusbbf 6G 5785 PTA- 12.5040, PTA- 12.5041, PT A-125042, PTA-125049, PTA-125050, PTA-125052 Ascusbbf 6176D 5382 PTA- 125049.PTA- 125051, PTA- 125052 Ascusbbf 107121 5586 PTA- 125042, PTA-125050 Ascusbbf 6H 5786 PTA- 125041, PTA- 125042, PTA- 125050 Ascusbbf 6176E 5383 PTA- 125049.PTA- Ascusbbf 6012A 5587 PTA-124942 Ascusbbf 61 5787 PTA-125041,PTA- 2026204768   19 Jun 2026 125051, PTA- 125052 125050 Ascusbbf 6176F 5384 PTA-125049 Ascusbbf 6012B 5588 PTA- 125040, PTA- 125041, PTA- 125049, PTA- 125050 Ascusbbf 6J 5788 PTA-125041, PTA-125050 Ascusbbf 6176G 5385 PTA- 125049, PTA- 125050, PTA- 125051.PTA- 125052 Ascusbbf 6012C 5589 PTA- 125040, PTA- 125041, PTA- 125049, PTA- 125050 Ascusbbf 6K 5789 PTA-125042 Ascusbbf 6176H 5386 PTA-125049,PTA-125051.PTA-125052 Ascusbbf 6012D 5590 PTA- 125041,PTA-125049 Ascusbbf 6L 5790 PTA-125042 Ascusbbf 61761 5387 PTA-125050.PTA-125051,PTA-125052 Ascusbbf 6012E 5591 PTA-125041 Ascusbbf 19022 A 5791 PTA- 125033, PTA-125040,PTA- 125041 Ascusbbf 4883A 5388 PTA- 12 5042,PTA-125049 Ascusbbf 2297A 5592 PTA- 125049, PTA- 125050, PTA- 125051 Ascusbbf 19022 B 5792 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125050 Ascusbbf 4883B 5389 PTA- 125049, PTA- 125051 Ascusbbf 2297B 5593 PTA- 125049, PTA- 125051 Ascusbbf 19022 C 5793 PTA- 125042, PTA-125052 Ascusbbf 4883C 5390 PTA- 125049, PTA- 125050, PTA- 125051 Ascusbbf 2297C 5594 PTA- 125049, PTA- 125051, PTA- 12.5052 Ascusbbf 2624A 5794 PTA- 125033, PTA- 125041, PTA- 125042, PTA- 125051, PTA- 125052 Ascusbbf 4883D 5391 PTA- 125049, PTA- Ascusbbf 2297D 5595 PTA-125049,PTA- Ascusbbf 2624B 5795 PTA-125041,PTA- 2026204768   19 Jun 2026 125050, PTA- 125051 125051, PTA- 125052 125042,PTA- 125050,PTA- 125051,PTA-125052 Ascusbbf 4883E 5392 PTA-125050 Ascusbbf 2297E 5596 PTA- 125049, PTA- 125051, PTA- 125052 Ascusbbf 2624C 5796 PTA- 125042, PTA- 125051, PTA- 125052 Ascusbbf_13543 A 5393 PTA-125033 Ascusbbf 2297F 5597 PTA- 125051, PTA- 125052 Ascusbbf 2624D 5797 PTA- 125042, PTA- 125051, PTA- 125052 Ascusbbf 13543B 5394 PTA-125041,PTA-125050 Ascusbbf 2297G 5598 PTA- 125051, PTA- 125052 Ascusbbf 2624E 5798 PTA- 125042, PTA- 125051, PTA- 125052 Ascusbbf 13543C 5395 PTA- 125041, PTA- 12 5042, PTA- 125050, PTA-125051 Ascusbbf 2297H 5599 PTA-125051 Ascusbbf 2624F 5799 PTA- 125051, PTA- 125052 Ascusbbf 13543 D 5396 PTA- 12 5041,PTA-125050 Ascusbbf 873A 5600 PTA-125033 Ascusbbf 2624G 5800 PTA-125051 Ascusbbf 13543E 5397 PTA-125033 ,PTA-125041 ,PTA-125042,PTA-125051, PTA-125052 Ascusbbf 873B 5601 PTA- 125033, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA-12.5052 Ascusbbf 3427A 5801 PTA- 125041, PTA- 125049,PTA-125050 Ascusbbf 152 A 5398 PTA-125051 Ascusbbf 873C 5602 PTA- 125040, PTA- 12.5041, PT A- 12.5042, PTA-125050 Ascusbbf 3427B 5802 PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 2026204768   19 Jun 2026 125051, PTA- 125052 Ascusbbf 152B 5399 PTA-125051 Ascusbbf 873D 5603 PTA- 125041, PTA- 125042, PTA- 125050 Ascusbbf 3427C 5803 PTA-125042, PTA-125049, PTA-125050,PTA-125051 Ascusbbf 152C 5400 PTA-125051 Ascusbbf 873E 5604 PTA- 125033, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050 Ascusbbf 3427D 5804 PTA-125050 Ascusbbf 707A 5401 PTA-125049,PTA-125050, PTA-125051, PTA-125052 Ascusbbf 873F 5605 PTA- 125033, PTA- 125041, PTA- 125042, PTA-12.5052 Ascusbbf 3427E 5805 PTA-125052 Ascusbbf 707B 5402 PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 873G 5606 PTA-125042 Ascusbbf 5005A 5806 PTA- 125049,PTA- 125050,PTA-125051 Ascusbbf 707C 5403 PTA-125049, PTA-125050 Ascusbbf 269A 5607 PTA- 125033, PTA- 12.5041, PTA- 125042, PTA- 125050 Ascusbbf 5005B 5807 PTA-125050 Ascusbbf 707D 5404 PTA- 125049, PTA- 125051, PTA- 125052 Ascusbbf 269B 5608 PTA-125033 Ascusbbf 69A 5808 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA-125049,PTA-125050,PTA-125051,PTA- 2026204768   19 Jun 2026 125052 Ascusbbf 707E 5405 PTA- 125049, PTA-125051,PTA-125052 Ascusbbf 269C 5609 PTA- 125033, PTA- 12.5042 Ascusbbf 69B 5809 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 707F 5406 PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 269D 5610 PTA-125041, PTA-125050 Ascusbbf 69C 5810 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 707G 5407 PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 269E 5611 PTA- 125033, PTA- 125041, PTA- 125050 Ascusbbf 69D 5811 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 707H 5408 PTA- 125049.PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 269F 5612 PTA-125033 Ascusbbf 69E 5812 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 12.5042, PTA- 12.5049, PTA- 125050, PTA- 125051, PTA-125052 2026204768   19 Jun 2026 Ascusbbf 707T 5409 PTA-125051 Ascusbbf 269G 5613 PTA-125033 Ascusbbf 69F 5813 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 707J 5410 PTA-125051 Ascusbbf 41015A 5614 PTA- 125033, PTA- 125041, PTA- 125050 Ascusbbf 69G 5814 PTA- 125033, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 1238A 5411 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 41015B 5615 PTA-125033 Ascusbbf 69H 5815 PTA- 125033,PTA- 125041,PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA-12.5052 Ascusbbf 1238B 5412 PTA- 125040, PTA-125041.PTA- 125050, PTA-125052 Ascusbbf 4J015C 5616 PTA-125041 Ascusbbf 691 5816 PTA-125033,PTA-125041,PT A-12.5042, PTA-12.5049, PTA-125051, PTA-125052 Ascusbbf 1238C 5413 PTA- 125033, PTA- 125040,PTA- 125041.PTA- Ascusbbf 4J015D 5617 PTA-125042 Ascusbbf 69J 5817 PTA-125033,PTA-125042,PTA-125051,PTA- 2026204768   19 Jun 2026 125042 ,PT A- 125050, PTA- 125051, PTA-125052 125052 Ascusbbf 1238D 5414 PTA- 125033 ,PTA-125051, PTA-125052 Ascusbbf 41015b 5618 PTA-125042 Ascusbbf 8082A 5818 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125050, PTA- 125052 Ascusbbf 5588A 5415 PTA- 125040, PTA- 125041, PTA- 125042, PTA-125049, PTA-125050 Ascusbbf 32877A 5619 PTA-124942 Ascusbbf 8082B 5819 PTA- 125040, PTA- 125041, PTA- 125042, PTA-125050 Ascusbbf 5588B 5416 PTA-125040,PTA-125042 Ascusbbf 57294A 5620 PTA-124942, PTA-125041 Ascusbbf 8082C 5820 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125050, PTA- 125052 Ascusbbf 5588C 5417 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051 Ascusbbf 57294B 5621 PTA- 125033, PTA-125050 Ascusbbf 8082D 5821 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125050, PTA- 125052 Ascusbbf 5588D 5418 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049.PTA- Ascusbbf 57294C 5622 PTA- 125033, PTA- 125052 Ascusbbf 8082E 5822 PTA- 125033, PTA- 12.5041 2026204768   19 Jun 2026 125050, PTA- 125051, PTA- 125052 Ascusbbf 5588E 5419 PTA-125042 Ascusbbf 27932A 5623 PTA-125040 Ascusbbf 8082 F 5823 PTA- 125033, PTA-125041,PTA- 125042 Ascusbbf 5588F 5420 PTA-125042 Ascusbbf 27932B 5624 PTA- 125040, PTA- 125041, PTA- 125050 Ascusbbf 8082 G 5824 PTA-125033,PTA-125042 Ascusbbf 5588G 5421 PTA-125042 Ascusbbf 27932C 5625 PTA- 125040, PTA- 125049, PTA- 125052 Ascusbbf 8082 H 5825 PTA-125033,PTA-125042 Ascusbbf 5588II 5422 PTA- 125049, PTA- 125051 Ascusbbf 22558A 5626 PTA-125051 Ascusbbf 80821 5826 PTA-125033 Ascusbbf 4691A 5423 PTA-125050.PTA-125051 Ascusbbf 22558B 5627 PTA-125051 Ascusbbf 95A 5827 PTA-125049, PTA-125050 Ascusbbf 4691B 5424 PTA- 125051, PTA- 125052 Ascusbbf_983757 A 5628 PTA- 125051, PTA- 125052 Ascusbbf 95B 5828 PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 469IC 5425 PTA-125051 Ascusbbf_983757 B 5629 PTA- 125051, PTA- 125052 Ascusbbf 95C 5829 PTA-125049, PTA-125050 Ascusbbf 9770A 5426 PTA-125041.PTA-125042, PTA-125049, PTA-125050 Ascusbbf_983757 C 5630 PTA- 125051, PTA- 125052 Ascusbbf 95D 5830 PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 9770B 5427 PTA- 125041.PTA- Ascusbbf 52330A 5631 PTA-125033 Ascusbbf 95E 5831 PTA-125051,PTA- 2026204768   19 Jun 2026 125049, PTA- 125050 125052 Ascusbbf 9770C 5428 PTA- 125049, PTA- 125051, PTA- 125052 Ascusbbf 1085A 5632 PTA- 124942, PTA- 125033, PTA- 125041, PTA- 125049, PTA- 125050 Ascusbbf 95F 5832 PTA-125051, PTA-125052 Ascusbbf 9770D 5429 PTA- 125049, PTA- 125050 Ascusbbf 1085B 5633 PTA- 125033, PTA- 125042, PTA- 125050, PTA- 125051, B- 67554 Ascusbbf 95G 5833 PTA- 125051, PTA- 125052. Ascusbbf 9770E 5430 PTA- 125049, PTA-125050 Ascusbbf 1085C 5634 PTA- 125033, PTA- 125040, PTA- 12.5041, PT A- 12.5042, PTA- 12.5049, PTA- 125050, PTA- 125051, PTA-125052 Ascusbbf 9511 5834 PTA-125051 Ascusbbf 9770F 5431 PTA- 125049, PTA- 125050 Ascusbbf 1085D 5635 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125052 Ascusbbf 1136A 5835 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050 Ascusbbf 9770G 5432 PTA- 125049, PTA- 125050 Ascusbbf 1085E 5636 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- Ascusbbf 1136B 5836 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 12.5050,PTA- 2026204768   19 Jun 2026 125050, PTA- 125052 125052 Ascusbbf 9770H 5433 PTA- 125049, PTA- 125050, PTA- 125051 Ascusbbf 1085F 5637 PTA- 125033, PTA- 125049, PTA- 125051, PTA- 125052 Ascusbbf 1136C 5837 PTA-125033, PTA-125040,PTA-125041,PTA-125042,PTA-125049, PTA-125050 Ascusbbf 14146 A     " 5434 PTA- 124942 ,PTA- 125033 ,PTA- 125041 ,PTA- 125042, PTA- 125051, PTA- 125052, B- 67555 Ascusbbf 1085G 5638 PTA-12.5033, PTA-125050 Ascusbbf 1136D 5838 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125052 Ascusbbf 14146B 5435 PTA- 125033, PTA- 125041, PTA- 125042, PTA- 125051, PTA- 125052 Ascusbbf 1085H 5639 PTA-125033, PTA-125050 Ascusbbf 1136E 5839 PTA- 125040, PTA- 125041, PTA- 125049, PTA- 125050 Ascusbbf 14J46C 5436 PTA-125041 Ascusbbf 10851 5640 PTA-125033, PTA-125050 Ascusbbf 1136F 5840 PTA-125042 Ascusbbf_14146 D 5437 PTA- 125033, PTA- 125041, PTA- 125042, PTA- 125051, PTA- 125052 Ascusbbf 1085J 5641 PTA-125033, PTA-125042 Ascusbbf 2770A 5841 PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051 Ascusbbf 14146E 5438 PTA-125033,PTA-125041 Ascusbbf 1085K 5642 PTA-125042 Ascusbbf 2770B 5842 PTA- 125049, PTA- 125050 Ascusbbf 14146F 5439 PTA-125041 Ascusbbf 154A 5643 PTA-124942 Ascusbbf 2770C 5843 PTA-125049 2026204768   19 Jun 2026 Ascusbbf 14146 G     “ 5440 PTA-125042,PTA-125051,PTA-125052 Ascusbbf 154B 5644 PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, B- 67550 Ascusbbf 1629A 5844 PTA-125033 Ascusbbf 1103A 5441 PTA- 125033,PTA-125041 Ascusbbf 154D 5645 PTA- 125040, PTA- 125041, PTA- 12.5042, PTA- 12.5051 Ascusbbf 1629B 5845 PTA- 125040, PTA- 125041 Ascusbbf 1103B 5442 PTA-125041, PTA-125051, PTA-125052 Ascusbbf 154E 5646 PTA- 125040, PTA- 12.5041, PT A- 12.5049, PTA- 12.5050, PTA-125052 Ascusbbf 1629C 5846 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050 Ascusbbf 1103C 5443 PTA-125041, PTA-125051, PTA-125052 Ascusbbf 154F 5647 PTA-12.5033, PTA-12.5040, PTA-125041, PTA-125042, PTA-125049, PTA-125050, PTA-125052 Ascusbbf 1629D 5847 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125049, PTA- 125050 Ascusbbf 1103D 5444 PTA- 125041, PTA- 125042, PTA- 125051 Ascusbbf 154G 5648 PTA- 125033, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA-125052. Ascusbbf 1629E 5848 PTA- 12.5041, PT A- 12.5050 Ascusbbf 1103E 5445 PTA- 125041, PTA- 125051.PTA- Ascusbbf 154H 5649 PTA- 125049, PTA- 125050 Ascusbbf 1629F 5849 PTA-125050 2026204768   19 Jun 2026 125052 Ascusbbf 1103F 5446 PTA- 125051, PTA- 125052 Ascusbbf 1541 5650 PTA- 125049, PTA- 125050 Ascusbbf 1629G 5850 PTA-125050 Ascusbbf 1103G 5447 PTA- 125051.PTA- 125052 Ascusbbf 154M 5651 PTA-125042 Ascusbbf 1821A 5851 PTA-125049 Ascusbbf 1103H 5448 PTA- 125051.PTA- 125052 Ascusbbf 1010A 5652 PTA-124942 Ascusbbf 182 IB 5852 PTA- 12.5049,PTA-125050 Ascusbbf 11031 5449 PTA-125051 Ascusbbf 1010B 5653 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 182 IC 5853 PTA-125049,PTA-125050 Ascusbbf 13717 A     " 5450 PTA- 125051, PTA- 125052 Ascusbbf 1010C 5654 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA-12.5052 Ascusbbf 182 ID 5854 PTA-125049,PTA-125050 Ascusbbf 13717B 5451 PTA-125051 Ascusbbf 1010D 5655 PTA- 125033, PTA- 12.5040, PTA- 12.5041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf_56782 A 5855 PTA- 125033, PTA-125041,PTA-125042,PTA-125049,PTA-125050,PTA-125051 2026204768   19 Jun 2026 Ascusbbf 13717C 5452 PTA-125051 Ascusbbf 1010E 5656 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA-12.5052 Ascusbbf 56782 B 5856 PTA-125051 Ascusbbf 876A 5453 PTA- 124942 ,PTA-125042 Ascusbbf 1010F 5657 PTA- 125033, PTA- 125040, PTA- 12.5041, PT A- 12.5052 Ascusbbf 56782 5857 PTA-125051 Ascusbbf 876B 5454 PTA-125042 Ascusbbf 1010G 5658 PTA- 125033, PTA-12.5040, PTA-12.5041, PTA-125042, PTA-125049, PTA-125050, PTA- 125051, PTA-125052 Ascusbbf 92A 5858 PTA- 125040, PTA-125050 Ascusbbf 876C 5455 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125050 Ascusbbf 1010H 5659 PTA-125033, PTA-125041 Ascusbbf 92B 5859 PTA- 125040, PTA- 125041, PTA- 125049, PTA- 125050 Ascusbbf 876D 5456 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125050, PTA- 125052 Ascusbbf 10101 5660 PTA- 125033, PTA-125041,PTA-125051 Ascusbbf 92C 5860 PTA- 12.5040, PTA- 12.5050 Ascusbbf 876E 5457 B-67553 Ascusbbf 1010.1 5661 PTA- 125041, PTA- 125050,PTA- Ascusbbf 92D 5861 PTA- 125040, PTA- 12.5049,PTA- 2026204768   19 Jun 2026 125052 125050, PTA- 125052 Ascusbbf 876F 5458 PTA-125033.PTA-125040, PTA-125041, PTA-125042, PTA-125050, PTA-125052 Ascusbbf 5575A 5662 PTA-124942 Ascusbbf 92E 5862 PTA-125040 Ascusbbf 876G 5459 PTA-125033 ,PTA-125042 Ascusbbf 5575B 5663 PTA-124942 Ascusbbf 92F 5863 PTA-125040,PTA-125049,PTA-125050 Ascusbbf 4936A 5460 PTA-125041 ,PTA-125050 Ascusbbf 5575C 5664 PTA- 125033, PTA-125042,PTA-125050 Ascusbbf 92G 5984 PTA- 125041, PTA- 125042 Ascusbbf 4936B 5461 PTA-125041 Ascusbbf 5575D 5665 PTA- 125042,PTA-125050 Ascusbbf 92H 5864 PTA-125041,PTA-125049 Ascusbbf 113152 A 5462 PTA-125050 Ascusbbf 5575E 5666 PTA- 125033, PTA- 125042, PTA- 125049, PTA- 125052 Ascusbbf 921 5985 PTA-125041 Ascusbbf 9031A 5463 PTA-125049 Ascusbbf 5575F 5667 PTA- 125033, PTA-125042,PTA-125049 Ascusbbf 92J 5986 PTA-125041 Ascusbbf 903 IB 5464 PTA-125049 Ascusbbf 5575G 5668 PTA- 125033, PTA-125042,PTA-125049,PTA-125050 Ascusbbf 92K 5865 PTA- 125041, PTA- 125051, PTA- 125052 Ascusbbf 903IC 5465 PTA-125049 Ascusbbf 5575H 5669 PTA- 125033, PTA- Ascusbbf 92L 5866 PTA-125050 2026204768   19 Jun 2026 125042,PTA- 125050 Ascusbbf 903 ID 5466 PTA- 125049, PTA- 125050 Ascusbbf 775A 5670 PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 118A 5867 PTA-125049,PTA-125051 Ascusbbf 903 IE 5467 PTA- 125049, PTA- 125050 Ascusbbf 775B 5671 PTA-125051 Ascusbbf 11 SB 5868 PTA-125051, PTA-125052 Ascusbbf 903 IF 5468 PTA- 125049, PTA- 125050 Ascusbbf 243 02A 5672 PTA-125041,PTA-125050,B-67551 Ascusbbf 118C 5869 PTA-125051 Ascusbbf 9031G 5469 PTA-125050 Ascusbbf 24302B 5673 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125049 Ascusbbf 5429A 5870 PTA- 125040, PTA- 125041, PTA- 125049, PTA- 125050, PTA- 125052 Ascusbbf 11823 A 5470 PTA-125041 Ascusbbf 24302C 5674 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125051, PTA- 125052 Ascusbbf 542.9B 5871 PTA- 125040, PTA- 125041, PTA- 125049, PTA- 125050, PTA-125052. Ascusbbf 11823B 5471 PTA- 125041 ,PTA- 125042 ,PTA- 125049 Ascusbbf 243 02D 5675 PTA-125041, PTA-12.5049 Ascusbbf 5429C 5872 PTA- 125033, PTA-125042. Ascusbbf 11823C 5472 PTA-125050 Ascusbbf 2.43 02E 5676 PTA- 125033, PTA- 125041, PTA- 12.5052 Ascusbbf 542.9D 5873 PTA-125049 Ascusbbf 1007A 5473 PTA-125041 Ascusbbf 24302F 5677 PTA-125033, PTA- Ascusbbf 3 A 5874 PTA- 125033,PTA- 2026204768   19 Jun 2026 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125052 125040, PTA- 125041, PTA- 125049, PTA- 125050, PTA-125052 Ascusbbf 24422 A     " 5474 PTA- 125051, PTA- 125052 Ascusbbf 2.4302G 5678 PTA- 125033, PTA- 125041, PTA- 125049, PTA- 12.5050, PTA- 12.5051, PTA- 12.5052 Ascusbbf 3B 5875 PTA- 125040, PTA- 125041, PTA- 125049, PTA- 125050, PTA-125052. Ascusbbf 95 J A 5475 PTA- 124942 ,PTA-125042 Ascusbbf 243 02H 5679 PTA-12.5041, PT A-125052 Ascusbbf 3C 5876 PTA- 125040, PTA- 125041, PTA- 125049, PTA- 125050 Ascusbbf 95 IB 5476 PTA-125033, PTA-125040, PTA-125041, PTA-125042, PTA-125049, PTA-125050, PTA-125051, PTA-125052 Ascusbbf 243021 5680 PTA- 125041, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 3D 5877 PTA-125040 Ascusbbf 951C 5477 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA-125050.PTA- 125051, PTA-125052 Ascusbbf 24302.1 5681 PTA- 125051, PTA- 125052 Ascusbbf 3E 5878 PTA-125042 Ascusbbf 95 ID 5478 PTA- Ascusbbf 1A 5682 PTA- Ascusbbf 3F 5879 PTA-125042. 2026204768   19 Jun 2026 125033, PTA-125040,PTA- 125041, PTA-12 5042, PT A-125049 125040, PTA- 125041, PTA- 125042, PTA-125052 Ascusbbf 95IE 5479 PTA- 125033 ,PTA- 125042, PTA- 125051, PTA- 125052 Ascusbbf IB 5683 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 12.5049, PTA- 12.5050, PTA- 12.5052 Ascusbbf 3G 5880 PTA-125050 Ascusbbf 95 IF 5480 PTA-125033 Ascusbbf IC 5684 PTA- 12.5040, PTA- 12.5041, PT A- 12.5042, PTA-125050 Ascusbbf 10576 A    " 5881 PTA- 125049, PTA- 125050, PTA- 125051 Ascusbbf 951G 5481 PTA-125033 Ascusbbf ID 5685 PTA- 125033, PTA- 12.5040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf_10576 B 5882 PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 5699A 5482 PTA- 125049, PTA- 125050 Ascusbbf IE 5686 PTA- 125033, PTA- 125040, PTA- 125042 Ascusbbf 10576 C     " 5883 PTA- 125049, PTA- 125050, PTA- 125051 Ascusbbf 5699B 5483 PTA- 125049, PTA- 125050 Ascusbbf IF 5687 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125050, PTA- Ascusbbf 10576 D 5884 PTA-125049 2026204768   19 Jun 2026 125051,PTA- 125052 Ascusbbf 5699C 5484 PTA- 125049, PTA- 125050 Ascusbbf 1G 5688 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA-12.5052 Ascusbbf 10576 E 5885 PTA-125051 Ascusbbf 5699D 5485 PTA- 125050, PTA-125051 Ascusbbf 1H 5689 PTA- 125033, PTA- 125040, PTA- 12.5041, PT A- 12.5050 Ascusbbf 729A 5886 PTA- 125049, PTA- 125051 Ascusbbf 130 A 5486 PTA- 125049, PTA- 125050, PTA- 125051, PTA-125052 Ascusbbf 11 5690 PTA- 125033, PTA- 12.5040, PTA- 12.5041, PT A- 12.5042, PTA- 125050, PTA- 125052 Ascusbbf 729B 5887 PTA- 125051, PTA- 125052 Ascusbbf 130B 5487 PTA-125049 Ascusbbf 1J 5691 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050 Ascusbbf 729C 5888 PTA-125051 Ascusbbf 130C 5488 PTA- 125049, PTA- 125050 Ascusbbf IK 5692 PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050 Ascusbbf 729D 5889 PTA-125051 Ascusbbf 130D 5489 PTA-125049,PTA- Ascusbbf 52548A 5693 PTA-125051,PTA- Ascusbbf 729E 5890 PTA-125051 2026204768   19 Jun 2026 125050 125052 Ascusbbf 130E 5490 PTA-125049 Ascusbbf 52548B 5694 PTA- 125051, PTA- 125052 Ascusbbf 729F 5891 PTA-125051 Ascusbbf 130F 5491 PTA- 125049.PTA- 125050 Ascusbbf 50658A 5695 PTA-125050 Ascusbbf 201A 5892 PTA-125042 Ascusbbf 1300 5492 PTA-125051 Ascusbbf 850A 5696 PTA-124942 Ascusbbf 201B 5893 PTA-12.5041,PTA-125042,PTA-125050,PTA-125051, PTA-125052 Ascusbbf 10109 A.       ” 5493 PTA-124942 Ascusbbf 850B 5697 PTA-125033 Ascusbbf 201C 5894 PTA-125033 Ascusbbf 10109E 5494 PTA-124942 Ascusbbf 850C 5698 PTA- 125033, PTA- 125040, PTA-125041,PTA-125049,PTA-125050 Ascusbbf 201D 5895 PTA-125033, PTA-12.5041,PTA-125042,PTA-125051,PTA-125052 Ascusbbf 10109C 5495 PTA-125033,PTA-125049 Ascusbbf 850D 5699 PTA-125040 Ascusbbf 201E 5896 PTA-125040, PTA-125041 Ascusbbf_10109 D 5496 PTA-125049 Ascusbbf 850E 5700 PTA- 125040, PTA- 125041, PTA- 125042,PTA-125049,PTA-125050 Ascusbbf 201F 5897 PTA-125040 Ascusbbf 10109E 5497 PTA- 125049, PTA- 125050 Ascusbbf 850F 5701 PTA-125033,PTA-125040,PTA-125041, PTA-125042, PTA-125049, PTA- Ascusbbf 201G 5898 PTA- 125033, PTA-125041,PTA- 125050 2026204768   19 Jun 2026 125050 Ascusbbf 10109F 5498 PTA-125049 Ascusbbf 850G 5702 PTA- 125033, PTA- 125040, PTA-125041,PTA-125049,PTA-125050 Ascusbbf 201H 5899 PTA-125033, PTA-125041, PTA-125042,PTA-125050 Ascusbbf 10109 G 5499 PTA- 125049, PTA- 125052 Ascusbbf 850H 5703 PTA- 125033, PTA-125041,PTA- 125050 Ascusbbf 2011 5900 PTA- 125033, PTA- 125041, PTA- 125042, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 10109 H 5500 PTA- 125049, PTA- 125050 Ascusbbf 8501 5704 PTA-125033,PTA-125050 Ascusbbf 416A 5901 PTA-125049 Ascusbbf 101091 5501 PTA-125050 Ascusbbf 850J 5705 PTA- 125033, PTA-125042 Ascusbbf 416B 5902 PTA- 125049, PTA- 125051, PTA- 125052 Ascusbbf 29797 A 5502 PTA-125051 Ascusbbf 4A 5706 PTA-124942 Ascusbbf 416C 5903 PTA-125049,PTA-125050 Ascusbbf 54068 A     " 5503 PTA-124942 Ascusbbf 4B 5707 PTA- 125040, PTA- 125041, PTA- 125042,PTA-125049,PTA- 125050,PTA-125052 Ascusbbf 416D 5904 PTA-125049,PTA-125050 Ascusbbf 54068B 5504 PTA-125033 Ascusbbf 4C 5708 PTA- 125041,PTA-125051 Ascusbbf 416E 5905 PTA-125049 Ascusbbf 54068C 5505 PTA-125033 ,PTA- Ascusbbf 4D 5709 PTA-125040, PTA- Ascusbbf 416F 5906 PTA-125049,PTA- 2026204768   19 Jun 2026 125040,PTA- 125041, PTA- 125042, PTA-125049 125041, PTA- 125042, PTA- 125050, PTA- 125051, PTA- 125052, B- 67552 125051 Ascusbbf 54068 D 5506 PTA- 125033 ,PTA- 125041 ,PTA- 125042, PTA- 125049, PTA- 125050 Ascusbbf 4E 5710 PTA- 125040, PTA- 12.5041, PT A- 12.5050, PTA- 12.5052 Ascusbbf 416G 5907 PTA-125050 Ascusbbf 7003A 5507 PTA-124942 Ascusbbf 4F 5711 PTA- 125033, PTA-12.5040, PTA- 12.5041, PT A-125042, PTA-125050, PTA-125051, PTA-125052 Ascusbbfl 5806 A 5908 PTA-125049 Ascusbbf 7003C 5508 PTA-125033 Ascusbbf 4G 5712 PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 15806 B 5909 PTA- 125049, PTA- 125051, PTA- 125052 Ascusbbf 23A 5509 PTA- 125051.PTA- 125052 Ascusbbf 4H 5713 PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 6115A 5910 PTA-125033, PTA-125041 Ascusbbf 1697A 5510 PTA-125049 Ascusbbf 41 5714 PTA-125041, PTA- Ascusbbf 6115B 5911 PTA- 12.5041,PT A- 2026204768   19 Jun 2026 125049, PTA- 125050, PTA- 125052 125050 Ascusbbf 1697B 5511 PTA-125050 Ascusbbf 4J 5715 PTA-125041,PTA-125050 Ascusbbf 6115C 5912 PTA-125041,PTA-125050 Ascusbbf 1697C 5512 PTA-125050 Ascusbbf 4K 5716 PTA-125051 Ascusbbf 6115D 5913 PTA-125041 Ascusbbf 7586A 5513 PTA-125041 Ascusbbf 5131A 5717 PTA- 125049, PTA- 125050, PTA- 125051 Ascusbbf 13250 58A 5914 PTA-125041,PTA-125049,PTA-125051 Ascusbbf 7586B 5514 PTA-125041, PTA-125042.PTA-125050 Ascusbbf 513 IB 5718 PTA-125049,PTA-125051 Ascusbbf 13250 58B    “ 5915 PTA-125041,PTA-125042,PTA-125049,PTA-125050,PTA-125051 Ascusbbf 7586C 5515 PTA- 125041, PTA- 125042.PTA- 125050 Ascusbbf 513 IC 5719 PTA-125049,PTA-125051 Ascusbbf 13250 58C   " 5916 PTA-125049 Ascusbbf 7586D 5516 PTA-125042 Ascusbbf 513 ID 5720 PTA-125049,PTA-125051 Ascusbbf 13250 58D 5917 PTA-125049 Ascusbbf 1034A 5517 PTA- 125051, PTA- 125052 Ascusbbf 513 IE 5721 PTA- 125051, PTA- 125052 Ascusbbf 13250 58E    " 5918 PTA-125050 Ascusbbf 1034B 5518 PTA-125051 Ascusbbf 513 IF 5722 PTA-125051 Ascusbbf 13250 58F 5919 PTA-125051 Ascusbbf 23134 A 5519 PTA-125049 Ascusbbf 5131G 5723 PTA-125051 Ascusbbf 28350 A 5920 PTA-125041,PTA-125050 Ascusbbf 43679 A    " 5520 PTA- 125040,PTA- 125041,PTA-125050 Ascusbbf 513IH 5724 PTA-125051 Ascusbbf 28350 B 5921 PTA-125041,PTA-125050 2026204768   19 Jun 2026 Ascusbbf 43679B 5521 PTA-125041 Ascusbbf 8600A 5725 PTA-125051 Ascusbbf_28350 C 5922 PTA-125041,PTA-125050 Ascusbbf 4367 9C 5522 PTA-125041 Ascusbbf 8600B 5726 PTA-125051 Ascusbbf 3 72A 5923 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125052 Ascusbbf 43679 D 5523 PTA-125041 Ascusbbf 1273A 5727' PTA-125049,PTA-125051,PTA-125052 Ascusbbf 372B 5924 PTA-125033 Ascusbbf 43679E 5524 PTA- 125041,PTA-125050 Ascusbbf 1273B 5728 PTA-125051 Ascusbbf 372C 5925 PTA-125040,PTA-125041,PTA- 125049,PTA-125050 Ascusbbf 43679F 5525 PTA-125050 Ascusbbf 1273C 5729 PTA-125051 Ascusbbf 372D 5926 PTA- 125033, PTA-125040,PTA-125041 Ascusbbf 63954 A     “ 5526 PTA-125049 Ascusbbf 1273D 5730 PTA-125051 Ascusbbf 372E 5927 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042,PTA-125049,PTA-125050 Ascusbbf 1517A 5527 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042,PTA- Ascusbbf 39159A 5731 PTA-125041 Ascusbbf 372F 5928 PTA-125033,PTA-125041,PTA-125050 2026204768   19 Jun 2026 125051, PTA- 125052 Ascusbbf 1517B 5528 PTA- 125040,PTA- 125041 ,PTA- 125042 ,PTA- 125051, PTA- 125052 Ascusbbf 39159B 5732 PTA-125041 Ascusbbf 372G 5929 PTA-125033,PTA-125042 Ascusbbf 1517C 5529 PTA- 125040, PTA- 125042 ,PTA- 125049, PTA- 125050, PTA- 125051 Ascusbbf 39159C 5733 PTA-125050 Ascusbbf 667A 5930 PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125052 Ascusbbf 15 GO 5530 PTA-125033 ,PTA-125040, PTA-125042 ,PTA-125049,PTA-125050, PTA-125051,PTA-125052 Ascusbbf 39159D 5734 PTA-125042 Ascusbbf 667B 5931 PTA-125042,PTA-125049 Ascusbbf 1517E 5531 PTA- 125033 ,PTA- 125041 ,PTA- 125050 Ascusbbf 318A 5735 PTA-125049,PTA-125050 Ascusbbf 1207A 5932 PTA- 125033, PTA- 125042, PTA- 125049, PTA- 125050 Ascusbbf 1517F 5532 PTA-125042 Ascusbbf 318B 5736 PTA-125049 Ascusbbf 1207B 5933 PTA- 125033,PTA-125042 Ascusbbf 1517G 5533 PTA-125042 Ascusbbf 318C 5737 PTA-125049,PTA-125050 Ascusbbf 1207C 5934 PTA-125033 Ascusbbf 1517H 5534 PTA-125042 Ascusbbf 318D 5738 PTA-125042,PTA-125050,PTA- Ascusbbf 1207D 5935 PTA- 125040,PTA-125041 2026204768   19 Jun 2026 125051 Ascusbbf 15171 5535 PTA-125049 Ascusbbf 318E 5739 PTA- 125042, PTA- 125050, PTA- 125051, PTA- 125052 Ascusbbf 1207E 5936 PTA-125040, PTA-125041 Ascusbbf 104A 5536 PTA-124942 Ascusbbf 7046A 5740 PTA-125050 Ascusbbf 1207F 5937 PTA- 125040, PTA- 125041, PTA- 125042 Ascusbbf 104B 5537 PTA- 125040, PTA- 12 5042, PTA- 125049, PTA-125050 Ascusbbf 137298 5 A     “ 5741 PTA-124942, PTA-12.5041 Ascusbbf 1207G 5938 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 125052 Ascusbbf 104C 5538 PTA- 125040, PTA- 12 5042, PTA- 125049, PTA- 125050 Ascusbbf 137298 5B 5742 PTA- 125033, PTA- 12.5041, PT A- 12.5042 Ascusbbf 1207H 5939 PTA- 125033,PTA-125040 Ascusbbf 104D 5539 PTA-125040 Ascusbbf 137298 5C 5743 PTA-125033, PTA-125041 Ascusbbf 12071 5940 PTA- 125033, PTA- 125040, PTA-125041,PTA-125049,PTA-125050 Ascusbbf 104E 5540 PTA- 125040, PTA- 125041, PTA- 125042 Ascusbbf 137298 5D 5744 PTA-125040, PTA-125041, PTA-125042,PTA-125049,PTA-125050 Ascusbbf 1207J 5941 PTA- 125033, PTA- 125042,PTA-125049 Ascusbbf 104F 5541 PTA- Ascusbbf 137298 5745 PTA- Ascusbbf 3875A. 5942 PTA- 2026204768   19 Jun 2026 125033, PTA- 125042 5E 125040, PTA- 125041, PTA- 125049,PTA-125050 125033,PTA- 125041,PTA-125042 Ascusbbf 104G 5542 PTA-125050 Ascusbbf! 37298 5F 5746 PTA-125041 Ascusbbf 3875B 5943 PTA-125040,PTA-125042 Ascusbbf 10411 5543 PTA-125042 Ascusbbf! 37298 5G 5980 PTA- 125033,PTA-125041 Ascusbbf 3875C 5944 PTA-125033,PTA-125041 Ascusbbf 1041 5544 PTA-125042 Ascusbbf 137298 511 5981 PTA- 125033,PTA-125041 Ascusbbf 3875D 5945 PTA-125041 Ascusbbf 148A 5545 PTA-125049, PTA-125051.PTA-125052 Ascusbbf 137298 51       ~ 5747 PTA- 125033,PTA-125041 Ascusbbf 72889 A     “ 5946 PTA- 125051,PTA- 125052 Ascusbbf 148B 5546 PTA- 125049, PTA- 125051 Ascusbbf! 37298 5J 5982 PTA- 125033,PTA- 125041,PTA- 125042,PTA- 125051, PTA- 125052 Ascusbbf_72889 B 5947 PTA-125051, PTA-125052 Ascusbbf 148C 5547 PTA-125049,PTA-125050, PTA-125051,PTA-125052 Ascusbbf 137298 5K    " 5983 PTA- 125033, PTA- 125041, PTA- 125050, PTA- 125051 Ascusbbf 10686 3A    ” 5948 PTA- 125051, PTA- 125052 Ascusbbf 148D 5548 PTA- 125051, PTA- 125052 Ascusbbf! 37298 5L 5748 PTA-125041 Ascusbbf_10686 3B 5949 PTA-125051 Ascusbbf 148E 5549 PTA- 125051, PTA- 125052 Ascusbbf! 37298 5M 5749 PTA-125041 Ascusbbf 120A 5987 PTA-125051 Ascusbbf 148F 5550 PTA- 125051.PTA- Ascusbbf 137298 5N    ” 5750 PTA-125041 Ascusbbf 120B 5950 PTA-125051 2026204768   19 Jun 2026 125052 Ascusbbf MSG 5551 PTA-125051 Ascusbbf 137298 50 5751 PTA-125041 Ascusbbf 120C 5951 PTA-125051 Ascusbbf 148H 5552 PTA-125051 Ascusbbf 137298 5P 5752 PTA-125041 Ascusbbf 1207K 5952 PTA-12.5033, PTA-125049, PTA-125050 Ascusbbf 944A 5553 PTA-124942 Ascusbbf 137298 5Q 5753 PTA-125041 Ascusbbf 1207L 5953 PTA-125049 Ascusbbf 944B 5554 PTA-125041,PTA- 125049, PTA- 125051.PTA- 125052 Ascusbbf 137298 5R 5754 PTA-125041 Ascusbbf 930A 5954 PTA-125051, PTA-125052 Ascusbbf 944C 5555 PTA- 125049,PTA-125051 Ascusbbf 137298 5S 5755 PTA- 125041, PTA- 125049, PTA- 125050 Ascusbbf 9 3 OB 5988 PTA-125051 Ascusbbf 944D 5556 PTA-125049.PTA-125050 Ascusbbf 137298 5T 5756 PTA- 125041, PTA- 125049, PTA- 125050 Ascusbbf 915 A 5955 PTA-125051 Ascusbbf 944E 5557 PTA-125049 Ascusbbf 121971 A 5757 PTA-125041 Ascusbbf 8941A 5956 PTA-125051, PTA-12.5052 Ascusbbf 944F 555S PTA-125049 Ascusbbf 52,51A 5758 PTA- 125033, PTA- 125049, PTA- 125050 Ascusbbf 8480A 5989 PTA-125051, PTA-12.5052 Ascusbbf 944G 5559 PTA-125049,PTA-125051,PTA-125052 Ascusbbf 525 IB 5759 PTA- 125033,PTA-125041 Ascusbbf 8480B 5957 PTA-125051 Ascusbbf 23033 A    " 5560 PTA-125051, PTA-125052 Ascusbbf 52,5IC 5760 PTA-125041, PTA-125049, PTA- Ascusbbf 3 74A 5990 PTA-125051, PTA-12.5052 2026204768   19 Jun 2026 125050,PTA- 125051 Ascusbbf 23033E 5561 PTA- 125051, PTA- 125052 Ascusbbf 525 ID 5761 PTA- 125033, PTA- 125041, PTA- 125049 Ascusbbf 374B 5958 PTA-125051, PTA-125052 Ascusbbf 23033C 5562 PTA- 125051, PTA- 125052 Ascusbbf 525 IE 5762 PTA- 125049, PTA- 125050,PTA-125051 Ascusbbf 374C 5991 PTA-125051, PTA-125052 Ascusbbf 23033 D 5563 PTA-125051 Ascusbbf 525 IF 5763 PTA- 125051, PTA- 125052 Ascusbbf 6906A 5959 PTA-125051, PTA-125052 Ascusbbf 2600A 5564 PTA-124942 Ascusbbf 5251G 5764 PTA-125051 Ascusbbf 6906B 5960 PTA-125051, PTA-125052 Ascusbbf 2600B 5565 PTA- 125033.PTA-125040,PTA-125041.PTA-125042 Ascusbbf 100 A 5765 PTA- 125033, PTA- 125049, PTA- 125050, PTA- 125052 Ascusbbf 6906C 5992 PTA-125051, PTA-125052 Ascusbbf 2600C 5566 PTA-125033, PTA-125040,PTA-125041, PTA-125042 Ascusbbf 100B 5766 PTA- 125040, PTA- 125042 Ascusbbf 6906D 5961 PTA-125051 Ascusbbf 2600D 556'7 PTA-125041 Ascusbbf 100C 5767' PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 125050, PTA- 12.5052 Ascusbbf 6906E 5962 PTA-125051 Ascusbbf 2600E 5568 PTA- 12 503 3,PTA- Ascusbbf 100D 5768 PTA- 125033, PTA- Ascusbbf 69K 5963 PTA- 125033,PTA- 2026204768   19 Jun 2026 125041 125040, PTA- 125041, PTA- 125050 125042 Ascusbbf 2600F 5569 PTA- 125033 ,PTA- 125041 Ascusbbf 100E 5769 PTA- 125033, PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA-12.5050 Ascusbbf 69L 5993 PTA- 125033,PTA-125042 Ascusbbf 2600G 5570 PTA-125041 Ascusbbf 100F 5770 PTA-125041 Ascusbbf 69M 5964 PTA-125033,PTA-125042 Ascusbbf 2600H 5571 PTA-125042 Ascusbbf 100G 5771 PTA-125042 Ascusbbf 69N 5965 PTA-125033,PTA-125042 Ascusbbf 8118A 5572 PTA- 125051, PTA- 125052 Ascusbbf 20584A 5772 PTA-125049 Ascusbbf 690 5966 PTA-125033,PTA-125042 Ascusbbf 8118B 5573 PTA-125051 Ascusbbf 205 84B 5773 PTA- 125051, PTA- 125052 Ascusbbf 69P 5967 PTA-125033 Ascusbbf 201A 5574 PTA-125042 Ascusbbf 4317A 5774 PTA-125041 Ascusbbf 721A 5968 PTA-125051, PTA-125052 Ascusbbf 201J 5575 PTA-125033.PTA-125042, PTA-125049 Ascusbbf 4317B 5775 PTA-125041 Ascusbbf 72 IB 5969 PTA-125051, PTA-125052 Ascusbbf 20 IK 5576 PTA-125042.PTA-125049, PTA-125051, PTA-125052 Ascusbbf 4317C 5776 PTA-125042 Ascusbbf 72IC 5970 PTA-125051, PTA-125052 Ascusbbf 201L 5577 PTA-125042.PTA- Ascusbbf 4317D 5777 PTA- 125051, PTA- Ascusbbf 3819A 5971 PTA-125051 2026204768   19 Jun 2026 125052 125052 Ascusbbf_10712 A 5578 PTA- 124942 ,PTA-125033.PTA-125042, PTA-125050 Ascusbbf 4317E 5778 PTA- 125051, PTA- 125052 Ascusbbf 4323A 5972 PTA-125051, PTA-125052 Ascusbbf 10712B 5579 PTA-124942 Ascusbbf 6A 5779 PTA- 125040, PTA- 125041, PTA- 125049, PTA- 125050, PTA- 125052 Ascusbbf 4323B 5973 PTA-125051, PTA-125052 Ascusbbf 10712C 5580 PTA- 125033 ,PTA-125042,PTA- 125049 Ascusbbf 6B 5780 PTA- 125040, PTA- 125041, PTA- 125042, PTA- 125049, PTA- 12.5050, PTA- 12.5052 Ascusbbf 4323C 5974 PTA- 125051, PTA- 125052. Ascusbbf 107!?. D 558! PTA- 125033 ,PTA- 125042 Ascusbbf 6C 5781 PTA- 125040, PTA- 12.5041, PT A- 12.5042, PTA- 125049, PTA- 125050, PTA-125052 Ascusbbf 4323D 5975 PTA-125051 Ascusbbf 10712E 5582 PTA-125042,PTA-125049 Ascusbbf 6D 5782 PTA- 125040, PTA- 125041, PTA- 125050, PTA- 125052 Ascusbbf 6087A 5976 PTA- 125051, PTA- 125052 Ascusbbf 6087B 5977 PTA- 125051, PTA- 125052 Ascusbbf 6087C 5978 PTA-125051 Ascusbbf 8414A 5979 PTA-125051 2026204768   19 Jun 2026 BRIEF DESCRIPTION OF THE FIGURES

[00196] FIG. 1 shows a general workflow of one embodiment of the method for determining the absolute abundance of one or more active microorganism strains.

[00197] FIG. 2 shows a general workflow of one embodiment of a method for determining the co-occurrence of one or more, or two or more, active microorganism strains in a sample with one or more metadata (environmental) parameters, followed by leveraging cluster analysis and community detection methods on the network of determined relationships.

[00198] FIG. 3 depicts a diagram that exemplifies how the diet influences the production of volatile fatty acids which in turn modulate milk production, body condition, growth, etc. Reproduced from Moran, 2005. Tropical daily farming: feeding management for small holder dairy- farmers in the humic tropics (Chapter 5), Landlinks Press, 312 pp.

[00199] FIG. 4 depicts the large change in microbial alpha-diversity as an animal continues to be fed a finishing diet.

[00200] FIG. 5 depicts the importance of the speed of microbial successions into a streamlined microbial ensemble related to animal efficiency. Furthermore, the large shift in the rumen microbiome on week 4 is most significant when related to efficiency.

[00201] FIG. 6 depicts the average ruminal VFA concentrations of high-RFI and low-RFI steers. More efficient animals tended to have a higher production of VFAs.

[00202] FIG. 7 depicts the theoretical concentrations of dissolved CO2 with respect to rumen pH. Reproduced from Laporte-Uribe, 2016.

[00203] FIG. 8 depicts the theorized pH of the rumen based on both VFA concentrations and dCO2 concentrations (filled squares), and VFA concentrations alone (open squares). Including dCO?. better predicts pH than VFAs alone. Reproduced from Laporte-Uribe, 2016.

[00204] FIG. 9 depicts the potential path ruminal CO2 may follow to impact the physiology of a ruminant. Reproduced from Laporte-Uribe, 2016.

[00205] FIG. 10A and FIG. 10B depict the phylogenetic diversity of rumen bacterial communities across samples at week 1 and week 5 of the study (FIG. 10A) and across samples at week 7 and week 10 of the study (FIG. 10B). 2026204768   19 Jun 2026

[00206] FIG. 11A and FIG. 11B depict the machine learning prediction accuracy utilized in the study. The residual feed intake (RFI) is predictive of serum metabolic signature (FIG. 11 A) and the RFI is predictive of rumen microbiome signature (FIG. 11B).

[00207] FIG. 12 depicts the mean predictive compounds between low-RFI and high-RFI. The greater the abundance of glucose-1-phosphate and / or glucose-6-phosphate is indicative of high-RFI.

[00208] FIG. 13 depicts the serum metabolic signatures correlated to week 10 bacterial community composition.

[00209] FIG. 14A and FIG. 14B depict the week 10 rumen microbial community correlations to serum metabolome. The mean serum pantothenate abundance differs between low-RFI and high-RFI steers (FIG. 14A). The mean Flavobacteriia abundance is associated with high pantothenate abundance (FIG. 14B).

[00210] FIG. 15 depicts spectral coclustering of composite average of OTU over time, indicating major microbial successions throughout the study in the rumen microbial community.

[00211] FIG. 16 depicts a principal coordinate analysis (PCoA) based on Bray-Curtis distances among the ruminal bacterial communities throughout the study.

[00212] FIG. 17 depicts the relative abundance of three bacterial orders driving the shift in bacterial community composition throughout the 10-week trial. Shaded regions indicate SEM. These three order are important to RFI. A major microbial succession occurs in week 4.

[00213] FIG. ISA and FIG. 18B depicts the microbial differentiation over pH between animals on week 5 following the large microbial succession in the rumen on week 4. Rumen alphadiversity correlated with pH (Pearson R=0.44; P=0.0051) (FIG. ISA). Rumen pH is predictive of bacterial community structure at week 5 (R2=0.48; P=0.04) (FIG. 18B). PC = Principle Component.

[00214] FIG. 19 depicts the plot of the residual feed intake (RFI) of the 50 steers.

[00215] FIG. 20 depicts a principal coordinate analysis (PCoA) of all samples throughout the study. Each dot represents a rumen sample’s bacterial community, and the distance between dots represents the difference between those communities. The shading of each dot represents the time in weeks when the sample was taken. 2026204768   19 Jun 2026

[00216] FIG. 21 depicts a Kullback-Leibler (K-L) divergence. The K-L divergence is a method for determining the commotional distance between two sets (i.e., difference between two sample’s microbial communities). In FIG. 21 reveals the comparison as to how much the microbial community changes week to week between high RFI animals and low RFI animals. The figure indicates that in week 4 the amount of change in the microbial communities is important to the RFI.

[00217] FIG. 22 depicts the AUC for the residual feed intake. Random Forests machine learning was used to predict the RFI on any given week given the microbial community composition. The microbial community is most predictive of RFI at week four.

[00218] FIG. 23 depicts the abundance of three taxonomic orders by week separated by RFI, which indicates the taxa that are having a direct correlation with RFI and at what time. The figure shows that week four is important for Aeromonadales and Pasteurellales populations in relation to RFI.

[00219] FIG. 24 depicts a flow chart for the synthesis of Vitamin B or pantothenate, and the intermediate products produced to arrive at Acyl-CoA species. Reproduced from Basu and Blair. 2012. Nature Protocols. 7:1-11.

[00220] FIG. 25 depicts the evaluation of the microbes of the present disclosure and their ability to modulate weight in the animal, particularly the ability to increase the weight of the animal.

[00221] FIG. 26 depicts the evaluation of the microbes of the present disclosure and their ability to modulate pH in the rumen, particularly the ability' to increase the pH in the rumen while the animal is being fed a high grain diet.

[00222] FIG. 27 depicts the evaluation of the microbes of the present disclosure and their ability to decrease CO2 in the rumen.

[00223] FIG. 28 depicts the evaluation of the microbes of the present disclosure and their ability to modulate CO2 in the rumen, particularly the ability to decrease CO2 in the rumen while the animal is being fed a high gram diet.

[00224] FIG. 29 depicts the MIC score distribution for common performance parameters including average daily weight gam, weight gain, feed intake, and feed efficiency with six 2026204768   19 Jun 2026 species of bacteria, in which many of the species have been evaluated in 3rd party studies. The lower the MIC score, the less likely the species / strams associated with that MIC score are capable of positively modulating the aforementioned performance parameters.

[00225] FIG. 30 depicts the KEGG carbon dioxide fixation pathways in prokaryotes.

[00226] FIG. 31 depicts the PATRIC carbon dioxide fixation pathways in prokaryotes.

[00227] FIG. 32 depicts the feed conversion ratio (FCR) for animals that received microbes vs. animals that did not receive microbes. DETAILED DESCRIPTION Definitions

[00228] While the following terms are believed to be well understood by one of ordinary skill in the art, the following definitions are set forth to facilitate explanation of the presently disclosed subject matter.

[00229] The term “a” or “an” may refer to one or more of that entity, i.e. can refer to plural referents. As such, the terms “a” or “an”, “one or more” and “at least one” are used interchangeably herein. In addition, reference to “an element” by the indefinite article “a” or “an” does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there is one and only one of the elements.

[00230] Reference throughout this specification to “one embodiment”, “an embodiment”, “one aspect”, or “an aspect” means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present disclosure. Thus, the appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics can be combined in any suitable manner in one or more embodiments.

[00231] As used herein, in particular embodiments, the terms “about” or “approximately” when preceding a numerical value indicates the value plus or minus a range of 10%. 2026204768   19 Jun 2026

[00232] As used herein the terms “microorganism” or “microbe” should be taken broadly. These terms are used interchangeably and include, but are not limited to, the two prokaryotic domains, Bacteria and Archaea, eukaryotic fungi and protozoa, as well as viruses. In some embodiments, the disclosure refers to the “microbes” of Table 1 and / or Table 2, or the “microbes” incorporated by reference. This characterization can refer to not only the predicted taxonomic microbial identifiers of the table, but also the identified strains of the microbes listed in the table.

[00233] The term “microbial community” means a group of microbes comprising two or more species or strains. Unlike microbial ensemble, a microbial community7 does not have to be carrying out a common function, or does not have to be participating in, or leading to, or correlating with, a recognizable parameter, such as a phenotypic trait of interest (e.g. increased feed efficiency in beef cattle).

[00234] As used herein, “isolate,” “isolated,” “isolated microbe,” and like terms, are intended to mean that the one or more microorganisms has been separated from at least one of the materials with which it is associated in a particular environment (for example soil, water, animal tissue).

[00235] Microbes of the present disclosure may include spores and / or vegetative cells. In some embodiments, microbes of the present disclosure include microbes in a viable but non-culturable (VBNC) state, or a quiescent state. See Liao and Zhao (US Publication US2015267163A1). In some embodiments, microbes of the present disclosure include microbes in a biofilm. See Merritt et al. (U.S. Patent 7,427,408).

[00236] Thus, an “isolated microbe” does not exist in its naturally occurring environment; rather, it is through the various techniques described herein that the microbe has been removed from its natural setting and placed into a non-naturally occurring state of existence. Thus, the isolated strain or isolated microbe may exist as, for example, a biologically pure culture, or as spores (or other forms of the strain) in association with an acceptable carrier.

[00237] As used herein, “spore” or “spores” refer to structures produced by bacteria and fungi that are adapted for survival and dispersal. Spores are generally characterized as dormant structures; however, spores are capable of differentiation through the process of germination. Germination is the differentiation of spores into vegetative cells that are capable of metabolic 2026204768   19 Jun 2026 activity, growth, and reproduction. The germination of a single spore results in a single fungal or bacterial vegetative cell. Fungal spores are units of asexual reproduction, and in some cases are necessary structures in fungal life cycles. Bacterial spores are structures for surviving conditions that may ordinarily be nonconductive to the survival or growth of vegetative cells.

[00238] As used herein, “microbial composition” refers to a composition comprising one or more microbes of the present disclosure, wherein a microbial composition, in some embodiments, is administered to animals of the present disclosure.

[00239] As used herein, “carrier”, “acceptable carrier”, or “pharmaceutical carrier” refers to a diluent, adjuvant, excipient, or vehicle with which the compound is administered. Such carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin; such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Water or aqueous solution saline solutions and aqueous dextrose and glycerol solutions are preferably employed as carriers, in some embodiments as injectable solutions. In some embodiments, gelling agents are employed as carriers. Alternatively, the carrier can be a solid dosage form carrier, including but not limited to one or more of a binder (for compressed pills), a glidant, an encapsulating agent, a flavorant, and a colorant. The choice of carrier can be selected with regard to the intended route of administration and standard pharmaceutical practice. See Hardee and Baggo (1998. Development and Formulation of Veterinary Dosage Forms. 2nd Ed. CRC Press. 504 pg ); E.W. Martin (1970. Remington’s Pharmaceutical Sciences. 17th Ed. Mack Pub. Co.); and Blaser etal. (US Publication US20110280840A1).

[00240] In some aspects, carriers may be granular in structure, such as sand or sand particles. In further aspects, the carriers may be dry, as opposed to a moist or wet carrier. In some aspects, carriers can be nutritive substances and / or prebiotic substances selected from fructooligosaccharides, inulms, isomalto-oligosaccharides, lactitol, lactosucruse, lactulose, pyrodextrines, soy oligosaccharides, transgalacto-oligosaccharides, xylo-oligosaccharides, trace minerals, and vitamins. In some aspects, carriers can be in solid or liquid form. In some aspects, carriers can be zeolites, calcium carbonate, magnesium carbonate, silicon dioxide, ground corn, trehalose, chitosan, shellac, albumin, starch, skim-milk powder, sweet-whey powder, maltodextrin, lactose, and inulm. In some aspects, a carrier is water or physiological saline. 2026204768   19 Jun 2026

[00241] The term “bioensemble,” “microbial ensemble,” or “synthetic ensemble” refers to a composition comprising one or more active microbes identified by methods, systems, and / or apparatuses of the present disclosure and that do not naturally exist in a naturally occurring environment and / or at ratios or amounts that do not exist in nature. A bioensemble is a subset of a microbial community of individual microbial species, or strains of a species, which can be described as carrying out a common function, or can be described as participating in, or leading to, or correlating with, a recognizable parameter, such as a phenotypic trait of interest (e.g. increased feed efficiency in feedlot cattle). The bioensemble may comprise two or more species, or strains of a species, of microbes. In some instances, the microbes coexist within the community symbiotically.

[00242] In certain aspects of the disclosure, the isolated microbes exist as isolated and biologically pure cultures. It will be appreciated by one of skill in the art, that an isolated and biologically pure culture of a particular microbe, denotes that said culture is substantially free (within scientific reason) of other living organisms and contains only the individual microbe in question. The culture can contain varying concentrations of said microbe. The present disclosure notes that isolated and biologically pure microbes often “necessarily differ from less pure or impure materials.” See, e.g. In re Bergstrom, 427 F.2d 1394, (CCPA J 970)(discussing purified prostaglandins), see also. In re Bergy, 596 F.2d 952 (CCPA 1979)(discussing purified microbes), see also, Parke-Davis & Co. v. H.K. Mulford & Co., 1 89 F. 95 (S.D.N.Y. 1911) (Learned Hand discussing purified adrenaline), aff’d in part, rev’d in part, 196 F. 496 (2d Cir. 1912), each of which are incorporated herein by reference. Furthermore, in some aspects, the disclosure provides for certain quantitative measures of the concentration, or purity limitations, that must be found within an isolated and biologically pure microbial culture. The presence of these purity values, in certain embodiments, is a further attribute that distinguishes the presently disclosed microbes from those microbes existing in a natural state. See, e.g., Merck & Co. v. Olin Mathieson Chemical Corp., 253 F.2d 156 (4th Cir. 1958) (discussing purity limitations for vitamin B12 produced by microbes), incorporated herein by reference.

[00243] As used herein, “individual isolates” should be taken to mean a composition, or culture, comprising a predominance of a single genera, species, or strain, of microorganism, following separation from one or more other microorganisms. The phrase should not be taken to indicate the extent to which the microorganism has been isolated or purified. However, 2026204768   19 Jun 2026 “individual isolates” can comprise substantially only one genus, species, or strain, of microorganism.

[00244] As used herein, “microbiome” refers to the collection of microorganisms that inhabit the digestive tract or gastrointestinal tract of an animal (including the rumen if said animal is a ruminant) and the microorganism’s physical environment (i.e. the microbiome has a biotic and physical component). The microbiome is fluid and may be modulated by numerous naturally occurring and artificial conditions (e.g, change in diet, disease, antimicrobial agents, influx of additional microorganisms, etc.). The modulation of the microbiome of a rumen that can be achieved via administration of the compositions of the disclosure, can take the form of: (a) increasing or decreasing a particular Family, Genus, Species, or functional grouping of microbe (i.e. alteration of the biotic component of the rumen microbiome) and / or (b) increasing or decreasing volatile fatty acids in the rumen, increasing or decreasing rumen pH, increasing or decreasing any other physical parameter important for rumen health (i.e. alteration of the abiotic component of the rumen microbiome).

[00245] As used herein, “probiotic” refers to a substantially pure microbe (i.e., a single isolate) or a mixture of desired microbes, and may also include any additional components that can be administered to beef cattle for restoring microbiota. Probiotics or microbial inoculant compositions of the disclosure may be administered with an agent to allow the microbes to survive the environment of the gastrointestinal tract, i.e., to resist low pH and to grow in the gastrointestinal environment. In some embodiments, the present compositions (e.g., microbial compositions) are probiotics in some aspects.

[00246] As used herein, “prebiotic” refers to an agent that increases the number and / or activity of one or more desired microbes. Non-limiting examples of prebiotics that may be useful in the methods of the present disclosure include fructooligosaccharides (e.g., oligofructose, inulin, inulm-type fructans), galactooligosaccharides, ammo acids, alcohols, and mixtures thereof. See Ramirez-Farias etal. (2008. Br. J. Nutr. 4:1-10) and Pool-Zobel and Sauer (2007. J. Nutr. 137:2580-2584 and supplemental).

[00247] The term “growth medium” as used herein, is any medium which is suitable to support growth of a microbe. By way of example, the media may be natural or artificial including gastrin supplemental agar, LB media, blood serum, and tissue culture gels. It should be 2026204768   19 Jun 2026 appreciated that the media may be used alone or in combination with one or more other media. It may also be used with or without the addition of exogenous nutrients.

[00248] The term “relative abundance” as used herein, is the number or percentage of a microbe present in the gastrointestinal tract or other organ system, relative to the number or percentage of total microbes present in said tract or organ system. The relative abundance may also be determined for particular types of microbes such as bacteria, fungi, viruses, and / or protozoa, relative to the total number or percentage of bacteria, fungi, viruses, and / or protozoa present. In one embodiment, relative abundance is determined by PCR. In another embodiment, relative abundance is determined by colony forming unit assays (cfu) or plaque forming unit assays (pfu) performed on samples from the gastrointestinal tract or other organ system of interest.

[00249] The medium may be amended or enriched with additional compounds or components, for example, a component which may assist in the interaction and / or selection of specific groups of microorganisms. For example, antibiotics (such as penicillin) or stenlants (for example, quaternary ammonium salts and oxidizing agents) could be present and / or the physical conditions (such as salinity, nutrients (for example organic and inorganic minerals (such as phosphorus, nitrogenous salts, ammonia, potassium and micronutrients such as cobalt and magnesium), pH, and / or temperature), methionine, prebiotics, ionophores, and beta glucans could be amended.

[00250] As used herein, the term “ruminant” includes mammals that are capable of acquiring nutrients from plant-based food by fermenting it in a specialized stomach (rumen) prior to digestion, principally through microbial actions. Ruminants included cattle, goats, sheep, giraffes, yaks, deer, antelope, and others.

[00251] As used herein, the term “bovid” includes any member of family Bovidae, which include hoofed mammals such as antelope, sheep, goats, and cattle, among others.

[00252] As used herein, the term “steer” includes any member, species, variant, or hybrid of Bos indicus, Bos taurus indicus, or Bos taurus taunts. The term “steer” further includes reference to cow (mature female), steer (castrated male), heifer (immature female not having born offspring), bull (mature uncastrated male), and calve (immature males or females). 2026204768   19 Jun 2026

[00253] As used herein, the terms “beef cattle” and “feedlot cattle” are used synonymously to refer to cattle that are grown and utilized for the production of beef. Said cattle of the present disclosure include varieties such as the following: Africander, Angus, Aubrac, Barzona, Bazadaise, Beef Shorthorn, Beefalo, Beefmaster, Belgian Blue, Belmont Red, Belted Galloway, Black Angus, Blonde d’Aquitaine, Bonsmara, Boran, Bradford, Brahman, Brahmousm, Brangus, British White, Buelmgo, Canchim, Caracu, Charolais, Chianma, Composite, Corriente, Devon, Dexter, Drakensberger, Droughtmaster, English Longhorn, Galloway, Gelbvieh, Gloucester, Hays Converter, Hereford, Highland, Holstein, Hybridmaster, Limousin, Lincoln Red, Lowline, Luing, Maine-Anjou, Rouge des Pres, Marchigiana, Miniature Hereford, Mirandesa, Mongolian, Murray Grey, Nel ore, Nguni, Parthenais, Piemontese, Pmzgauer, Red Angus, Red Poll, Retinta, Romagnola, Salers, Sanganer, Santa Cruz, Santa Gertrudis, Senepol, Shetland, Simbrah, Simmental, South Devon, Speckle Park, Square Meaters, Sussex, Tarentaise, Texas Longhorn, Tuli, Wagyu, Watusi, Welsh Black, Whitebred Shorthorn, and Zebu; or hybrids and / or crosses thereof.

[00254] As used herein, “dairy cattle” or “daily cows” are used synonymously to refer to cows that are grown and utilized for the production of milk.

[00255] As used herein, “performance” should be taken to be increased weight gain, improved feed efficiency, improved residual feed intake, improved feed intake.

[00256] As used herein, “improved” should be taken broadly to encompass improvement of a characteristic of interest, as compared to a control group, or as compared to a known a verage quantity associated with the characteristic in question. For example, “improved” feed efficiency associated with application of a beneficial microbe, or microbial ensemble, of the disclosure can be demonstrated by comparing the feed efficiency of beef cattle treated by the microbes taught herein to the feed efficiency of beef cattle not treated. In the present disclosure, “improved” does not necessarily demand that the data be statistically significant (i.e. p < 0.05); rather, any quantifiable difference demonstrating that one value (e.g. the average treatment value) is different from another (e.g. the average control value) can rise to the level of “improved.”

[00257] As used herein, “inhibiting and suppressing” and like terms should not be construed to require complete inhibition or suppression, although this may be desired in some embodiments. 2026204768   19 Jun 2026

[00258] The term “marker” or “unique marker” as used herein is an indicator of unique microorganism type, microorganism strain or activity of a microorganism strain. A marker can be measured in biological samples and includes without limitation, a nucleic acid-based marker such as a ribosomal RNA gene, a peptide- or protein-based marker, and / or a metabolite or other small molecule marker.

[00259] The term “metabolite” as used herein is an intermediate or product of metabolism. A metabolite in one embodiment is a small molecule. Metabolites have various functions, including in fuel, structural, signaling, stimulatory and inhibitory effects on enzymes, as a cofactor to an enzyme, in defense, and in interactions with other organisms (such as pigments, odorants and pheromones). A primary metabolite is directly involved in normal growth, development and reproduction. A secondary metabolite is not directly involved in these processes but usually has an important ecological function. Examples of metabolites include but are not limited to antibiotics and pigments such as resins and terpenes, etc. Some antibiotics use primary' metabolites as precursors, such as actinomycin which is created from the primary metabolite, tryptophan. Metabolites, as used herein, include small, hydrophilic carbohydrates; large, hydrophobic lipids and complex natural compounds.

[00260] As used herein, the term “genotype” refers to the genetic makeup of an individual cell, cell culture, tissue, organism, or group of organisms.

[00261] As used herein, the term “allele(s)” means any of one or more alternative forms of a gene, all of which alleles relate to at least one trait or characteristic. In a diploid cell, the two alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes. Since the present disclosure, in embodiments, relates to QTLs, i.e. genomic regions that may comprise one or more genes or regulatory sequences, it is in some instances more accurate to refer to “haplotype” (i.e. an allele of a chromosomal segment) instead of “allele”, however, in those instances, the term “allele” should be understood to comprise the term “haplotype”. Alleles are considered identical when they express a similar phenotype. Differences in sequence are possible but not important as long as they do not influence phenotype.

[00262] As used herein, the term “locus” (loci plural) means a specific place or places or a site on a chromosome where for example a gene or genetic marker is found. 2026204768   19 Jun 2026

[00263] As used herein, the term “genetically linked” refers to two or more traits that are co-inherited at a high rate during breeding such that they are difficult to separate through crossing.

[00264] A “recombination” or “recombination event” as used herein refers to a chromosomal crossing over or independent assortment. The term “recombinant” refers to an organism having a new genetic makeup arising as a result of a recombination event.

[00265] As used herein, the term “molecular marker” or “genetic marker” refers to an indicator that is used in methods for visualizing differences in characteristics of nucleic acid sequences. Examples of such indicators are restriction fragment length polymorphism (RFLP) markers, amplified fragment length polymorphism (AFLP) markers, single nucleotide polymorphisms (SNPs), insertion mutations, microsatellite markers (SSRs), sequence-characterized amplified regions (SCARs), cleaved amplified polymorphic sequence (CAI’S) markers or isozyme markers or combinations of the markers described herein w’hich defines a specific genetic and chromosomal location. Markers further include polynucleotide sequences encoding 16S or 18S rRNA, and internal transcribed spacer (ITS) sequences, which are sequences found between small-subunit and large-subunit rRNA genes that have proven to be especially useful in elucidating relationships or distinctions among when compared against one another. Mapping of molecular markers in the vicinity of an allele is a procedure which can be performed by the average person skilled in molecular-biological techniques.

[00266] The primary structure of major rRNA subunit 16S comprise a particular combination of conserved, variable, and hypervariable regions that evolve at different rates and enable the resolution of both very ancient lineages such as domains, and more modern lineages such as genera. The secondary structure of the 16S subunit include approximately 50 helices which result in base pairing of about 67% of the residues. These highly conserved secondary structural features are of great functional importance and can be used to ensure positional homology in multiple sequence alignments and phylogenetic analysis. Over the previous few decades, the 16S rRNA gene has become the most sequenced taxonomic marker and is the cornerstone for the current systematic classification of bacteria and archaea (Yarza el al. 2014. Nature Rev. Micro. 12:635-45). 2026204768   19 Jun 2026

[00267] A sequence identity of 94.5% or lower for two 16S rRNA genes is strong evidence for distinct genera, 86.5% or lower is strong evidence for distinct families, 82% or lower is strong evidence for distinct orders, 78.5% is strong evidence for distinct classes, and 75% or lower is strong evidence for distinct phyla. The comparative analysis of 16S rRNA gene sequences enables the establishment of taxonomic thresholds that are useful not only for the classification of cultured microorganisms but also for the classification of the many environmental sequences. Yarza et al. 2014. Nature Rev. Micro. 12:635-45).

[00268] As used herein, the term “trait” refers to a characteristic or phenotype. For example, in the context of some embodiments of the present disclosure; efficiency of feed utilization, particularly with corn-intensive diets; amount of feces produced; susceptibility to gut pathogens; and a decrease in mortality rates; among others. Desirable traits may also include other characteristics, including but not limited to: an increase in weight; an increase in average daily weight gain; an increase of musculature; an increase of fatty’ acid concentration in the gastrointestinal tract; an improved efficiency in feed utilization and digestibility; an increase in polysaccharide and lignin degradation; an increase in fat, starch, and / or protein digestion; an increase in fatty acid concentration in the rumen; pH balance in the rumen, an increase in vitamin availability; an increase in mineral availability’; an increase m amino acid availability; a reduction in methane and / or nitrous oxide emissions; a reduction m manure production; an improved dry matter intake; an improved efficiency of nitrogen utilization; an improved efficiency of phosphorous utilization; an increased resistance to colonization of pathogenic microbes that colonize cattle; reduced mortality; increased production of antimicrobials; increased clearance of pathogenic microbes; increased resistance to colonization of pathogenic microbes that colonize cattle; increased resistance to colonization of pathogenic microbes that infect humans; reduced incidence of acidosis or bloat; increased meat marbling, increased or decreased red coloring of meat, increased or decreased texture / coarseness of meat; increased amount of USDA Prime, USDA Choice, and USDA Select quality meat per animal, increased in the number of animals producing USDA Prime, USDA Choice, and USDA Select quality meat; increase or reduced concentration or presence of volatile compounds in the meat; reduced prevalence of acidosis or bloat; reduced body temperature; and any combination thereof; wherein said increase or reduction is determined by comparing against an animal not having been administered said composition. 2026204768   19 Jun 2026

[00269] A trait may be inherited in a dominant or recessive manner, or in a partial or incomplete-dominant manner. A trait may be monogenic (i.e. determined by a single locus) or polygenic (i.e. determined by more than one locus) or may also result from the interaction of one or more genes with the environment.

[00270] In the context of this disclosure, traits may also result from the interaction of one or more beef cattle genes and one or more microorganism genes.

[00271] As used herein, the term “homozygous” means a genetic condition existing when two identical alleles reside at a specific locus, but are positioned individually on corresponding pairs of homologous chromosomes in the cell of a diploid organism. Conversely, as used herein, the term “heterozygous” means a genetic condition existing when two different alleles reside at a specific locus, but are positioned individually on corresponding pairs of homologous chromosomes in the cell of a diploid organism.

[00272] As used herein, the term “phenotype” refers to the observable characteristics of an individual cell, cell culture, organism (e.g., cattle), or group of organisms which results from the interaction between that individual’s genetic makeup (i.e., genotype) and the environment.

[00273] As used herein, the term “chimeric” or “recombinant” when describing a nucleic acid sequence or a protein sequence refers to a nucleic acid, or a protein sequence, that links at least two heterologous polynucleotides, or two heterologous polypeptides, into a single macromolecule, or that re-arranges one or more elements of at least one natural nucleic acid or protein sequence. For example, the term “recombinant” can refer to an artificial combination of two otherwise separated segments of sequence, e.g., by chemical synthesis or by the manipulation of isolated segments of nucleic acids by genetic engineering techniques.

[00274] As used herein, a “synthetic nucleotide sequence” or “synthetic polynucleotide sequence” is a nucleotide sequence that is not known to occur in nature or that is not naturally occurring. Generally, such a synthetic nucleotide sequence will comprise at least one nucleotide difference when compared to any other naturally occurring nucleotide sequence.

[00275] As used herein, the term “nucleic acid” refers to a polymeric form of nucleotides of any length, either ribonucleotides or deoxyribonucleotides, or analogs thereof. This term refers to the primary' structure of the molecule, and thus includes double- and single-stranded 2026204768   19 Jun 2026 DNA, as well as double- and single-stranded RNA. It also includes modified nucleic acids such as methylated and / or capped nucleic acids, nucleic acids containing modified bases, backbone modifications, and the like. The terms “nucleic acid” and “nucleotide sequence” are used interchangeably.

[00276] As used herein, the term “gene” refers to any segment of DNA associated with a biological function. Thus, genes include, but are not limited to, coding sequences and / or the regulatory' sequences required for their expression. Genes can also include non-expressed DNA segments that, for example, form recognition sequences for other proteins. Genes can be obtained from a variety of sources, including cloning from a source of interest or synthesizing from known or predicted sequence information, and may include sequences designed to have desired parameters.

[00277] As used herein, the term “homologous” or “homologue” or “ortholog” is known in the art and refers to related sequences that share a common ancestor or family member and are determined based on the degree of sequence identity. The terms “homology,” “homologous,” “substantially similar” and “corresponding substantially” are used interchangeably herein. They refer to nucleic acid fragments wherein changes in one or more nucleotide bases do not affect the ability of the nucleic acid fragment to mediate gene expression or produce a certain phenotype. These terms also refer to modifications of the nucleic acid fragments of the instant disclosure such as deletion or insertion of one or more nucleotides that do not substantially alter the functional properties of the resulting nucleic acid fragment relative to the initial, unmodified fragment. It is therefore understood, as those skilled in the art will appreciate, that the disclosure encompasses more than the specific exemplary sequences. These terms describe the relationship between a gene found in one species, subspecies, variety, cultivar or strain and the corresponding or equivalent gene in another species, subspecies, variety, cultivar or strain. For purposes of this disclosure homologous sequences are compared. “Homologous sequences” or “homologues” or “orthologs” are thought, believed, or known to be functionally related. A functional relationship may be indicated in any one of a number of ways, including, but not limited to: (a) degree of sequence identity and / or (b) the same or similar biological function. Preferably, both (a) and (b) are indicated. Homology can be determined using software programs readily available in the art, such as those discussed in Current Protocols in Molecular Biology (F.M. Ausubel et al., eds., 1987) Supplement 30, section 7.718, Table 7.71. Some alignment programs are MacVector 2026204768   19 Jun 2026 (Oxford Molecular Ltd, Oxford, U.K.), .ALIGN Plus (Scientific and Educational Software, Pennsylvania) and AlignX (Vector NTI, Invitrogen, Carlsbad, CA). Another alignment program is Sequencher (Gene Codes, .Ann .Arbor, Michigan), using default parameters.

[00278] As used herein, the term “nucleotide change” refers to, e.g., nucleotide substitution, deletion, and / or insertion, as is well understood in the art. For example, mutations contain alterations that produce silent substitutions, additions, or deletions, but do not alter the properties or activities of the encoded protein or how the proteins are made.

[00279] As used herein, the term “protein modification” refers to, e.g., amino acid substitution, amino acid modification, deletion, and / or insertion, as is well understood in the art.

[00280] As used herein, the term “at least a portion” or “fragment” of a nucleic acid or polypeptide means a portion having the minimal size characteristics of such sequences, or any larger fragment of the full length molecule, up to and including the full length molecule. A fragment of a polynucleotide of the disclosure may encode a biologically active portion of a genetic regulatory' element. A biologically active portion of a genetic regulatory' element can be prepared by isolating a portion of one of the polynucleotides of the disclosure that comprises the genetic regulatory element and assessing activity as described herein. Similarly, a portion of a polypeptide may be 4 ammo acids, 5 ammo acids, 6 amino acids, 7 amino acids, and so on, going up to the full length polypeptide. The length of the portion to be used will depend on the particular application. A portion of a nucleic acid useful as a hybridization probe may be as short as 12 nucleotides; in some embodiments, it is 20 nucleotides. A portion of a polypeptide useful as an epitope may be as short as 4 ammo acids. A portion of a polypeptide that performs the function of the full-length polypeptide would generally be longer than 4 ammo acids.

[00281] Variant polynucleotides also encompass sequences derived from a mutagenic and recombinogenic procedure such as DNA shuffling. Strategies for such DNA shuffling are known in the art. See, for example, Stemmer (1994) PNAS 91:10747-10751; Stemmer (1994) Nature 370:389-391; Crameri et al.(]997) Nature Biotech. 15:436-438; Moore e / « / .(1997) J. Mol, Biol. 272:336-347; Zhang et « / .(1997) PNAS 94:4504-4509; Crameri el « / .(1998) Nature 391:288291; and U.S. Patent Nos. 5,605,793 and 5,837,458. For PCR amplifications of the polynucleotides disclosed herein, oligonucleotide primers can be designed for use in PCR reactions to amplify corresponding DNA sequences from cDNA or genomic DNA extracted 2026204768   19 Jun 2026 from any organism of interest. Methods for designing PCR primers and PCR cloning are generally known in the art and are disclosed in Sambrook el al.(Y)39) Molecular Cloning: A Laboratory Manual (2nd ed., Cold Spring Harbor Laboratory Press, Plainview, New York). See also Innis et al., eds. (1990) PCR Protocols: A Guide to Methods and Applications (Academic Press, New York); Innis and Gelfand, eds. (1995) PCR Strategies (Academic Press, New York); and Innis and Gelfand, eds. (1999) PCR Methods Manual (Academic Press, New York). Known methods of PCR include, but are not limited to, methods using paired primers, nested primers, single specific primers, degenerate primers, gene-specific primers, vector-specific primers, partially-mismatched primers, and the like.

[00282] The term “primer” as used herein refers to an oligonucleotide which is capable of annealing to the amplification target allowing a DNA polymerase to attach, thereby serving as a point of initiation of DNA synthesis when placed under conditions in which synthesis of primer extension product is induced, i.e., in the presence of nucleotides and an agent for polymerization such as DNA polymerase and at a suitable temperature and pH. The (amplification) primer is preferably single stranded for maximum efficiency in amplification. Preferably, the primer is an oligodeoxyribonucleotide. The primer must be sufficiently long to prime the synthesis of extension products in the presence of the agent for polymerization. The exact lengths of the primers will depend on many factors, including temperature and composition (A / T vs. G / C content) of primer. A pair of bi-directional primers consists of one forward and one reverse primer as commonly used in the art of DNA amplification such as in PCR amplification.

[00283] The terms “stringency” or “stringent hybridization conditions” refer to hybridization conditions that affect the stability of hybrids, e.g., temperature, salt concentration, pH, formamide concentration and the like. These conditions are empirically optimized to maximize specific binding and minimize non-specific binding of primer or probe to its target nucleic acid sequence. The terms as used include reference to conditions under which a probe or primer will hybridize to its target sequence, to a detectably greater degree than other sequences (e.g. at least 2-fold over background). Stringent conditions are sequence dependent and will be different in different circumstances. Longer sequences hybridize specifically at higher temperatures. Generally, stringent conditions are selected to be about 5° C lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength and pH) at which 50% of a complementary target 2026204768   19 Jun 2026 sequence hybridizes to a perfectly matched probe or primer. Typically, stringent conditions will be those in which the salt concentration is less than about 1.0 M Na+ ion, typically about 0.01 to 1.0 M Na + ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30° C for short probes or primers (e.g. 10 to 50 nucleotides) and at least about 60° C for long probes or primers (e.g. greater than 50 nucleotides). Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide. Exemplary low stringent conditions or “conditions of reduced stringency” include hybridization with a buffer solution of 30% formamide, 1 M NaCl, 1% SDS at 37° C and a wash in 2 SSC at 40° C. Exemplary high stringency conditions include hybridization in 50% formamide, IM NaCl, 1% SDS at 37° C, and a wash in O.lxSSC at 60° C. Hybridization procedures are well known in the art and are described by e.g. Ausubel et al., 1998 and Sambrook et al., 2001. In some embodiments, stringent conditions are hybridization in 0.25 M Na2HPO4 buffer (pH 7.2) containing 1 mM Na2EDTA, 0.5-20% sodium dodecyl sulfate at 45°C, such as 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20%, followed by a wash in 5xSSC, containing 0.1% (w / v) sodium dodecyl sulfate, at 55°C to 65°C.

[00284] As used herein, “promoter” refers to a DNA sequence capable of controlling the expression of a coding sequence or functional RNA. The promoter sequence consists of proximal and more distal upstream elements, the latter elements often referred to as enhancers. Accordingly, an “enhancer” is a DNA sequence that can stimulate promoter activity, and may be an innate element of the promoter or a heterologous element inserted to enhance the level or tissue specificity of a promoter. Promoters may be derived in their entirety from a native gene, or be composed of different elements derived from different promoters found in nature, or even comprise synthetic DNA segments. It is understood by those skilled in the art that different promoters may direct the expression of a gene in different tissues or cell types, or at different stages of development, or in response to different environmental conditions. It is further recognized that since in most cases the exact boundaries of regulatory sequences have not been completely defined, DNA fragments of some variation may have identical promoter activity .

[00285] As used herein, a “constitutive promoter” is a promoter which is active under most conditions and / or during most development stages. There are several advantages to using constitutive promoters in expression vectors used in biotechnology, such as: high level of production of proteins used to select transgenic cells or organisms; high level of expression of 2026204768   19 Jun 2026 reporter proteins or scorable markers, allowing easy detection and quantification; high level of production of a transcription factor that is part of a regulatory transcription system; production of compounds that requires ubiquitous activity in the organism; and production of compounds that are required during all stages of development. Non-limiting exemplary constitutive promoters include, CaMV 35S promoter, opine promoters, ubiquitin promoter, alcohol dehydrogenase promoter, etc.

[00286] As used herein, a “non-constitutive promoter” is a promoter which is active under certain conditions, in certain types of cells, and / or during certain development stages. For example, tissue specific, tissue preferred, cell type specific, cell type preferred, inducible promoters, and promoters under development control are non-constitutive promoters. Examples of promoters under developmental control include promoters that preferentially initiate transcription in certain tissues.

[00287] As used herein, “inducible” or “repressible” promoter is a promoter which is under chemical or environmental factors control. Examples of environmental conditions that may affect transcription by inducible promoters include anaerobic conditions, certain chemicals, the presence of light, acidic or basic conditions, etc.

[00288] As used herein, a “tissue specific” promoter is a promoter that initiates transcription only in certain tissues. Unlike constitutive expression of genes, tissue-specific expression is the result of several interacting levels of gene regulation. As such, m the art sometimes it is preferable to use promoters from homologous or closely related species to achieve efficient and reliable expression of transgenes in particular tissues. This is one of the mam reasons for the large amount of tissue-specific promoters isolated from particular tissues found in both scientific and patent literature.

[00289] As used herein, the term “operably linked” refers to the association of nucleic acid sequences on a single nucleic acid fragment so that the function of one is regulated by the other. For example, a promoter is operably linked with a coding sequence when it is capable of regulating the expression of that coding sequence (i.e., that the coding sequence is under the transcriptional control of the promoter). Coding sequences can be operably linked to regulatory-sequences in a sense or antisense orientation. In another example, the complementary RNA regions of the disclosure can be operably linked, either directly or indirectly, 5' to the target 2026204768   19 Jun 2026 mRNA, or 3' to the target mRNA, or within the target mRNA, or a first complementary region is 5' and its complement is 3’ to the target mRNA.

[00290] As used herein, the phrases “recombinant construct”, “expression construct”, “chimeric construct”, “construct”, and “recombinant DNA construct” are used interchangeably herein. A recombinant construct comprises an artificial combination of nucleic acid fragments, e.g., regulatory and coding sequences that are not found together in nature. For example, a chimeric construct may comprise regulatory sequences and coding sequences that are derived from different sources, or regulatory sequences and coding sequences derived from the same source, but arranged in a manner different than that found in nature. Such construct may be used by itself or may be used in conjunction with a vector. If a vector is used then the choice of vector is dependent upon the method that will be used to transform host cells as is well known to those skilled in the art. For example, a plasmid vector can be used. The skilled artisan is well aware of the genetic elements that must be present on the vector in order to successfully transform, select and propagate host cells comprising any of the isolated nucleic acid fragments of the disclosure. The skilled artisan will also recognize that different independent transformation events will result in different levels and patterns of expression (Jones et al., (1985) EMBO J. 4:2411-2418; De Almeida et al., (1989) Mol. Gen. Genetics 218:78-86), and thus that multiple events must be screened in order to obtain lines displaying the desired expression level and pattern. Such screening may be accomplished by Southern analysis of DNA, Northern analysis of mRNA expression, immunoblotting analysis of protein expression, or phenotypic analysis, among others. Vectors can be plasmids, viruses, bacteriophages, proviruses, phagemids, transposons, artificial chromosomes, and the like, that replicate autonomously or can integrate into a chromosome of a host cell. A vector can also be a naked RNA polynucleotide, a naked DNA polynucleotide, a polynucleotide composed of both DNA and RNA within the same strand, a po1y-lysine-conjugated DNA or RNA, a peptide-conjugated DN A or RN A, a liposome-conjugated DN A, or the like, that is not autonomously replicating. As used herein, the term “expression” refers to the production of a functional end-product e.g, an mRNA or a protein (precursor or mature).

[00291] In some embodiments, the cell or organism has at least one heterologous trait. As used herein, the term “heterologous trait” refers to a phenotype imparted to a transformed host cell or transgenic organism by an exogenous DNA segment, heterologous polynucleotide or 2026204768   19 Jun 2026 heterologous nucleic acid. Various changes in phenotype are of interest to the present disclosure, including but not limited to increasing yield of an economically important trait (e.g., weight, etc.) and the like. These results can be achieved by providing expression of heterologous products or increased expression of endogenous products in organisms using the methods and compositions of the present disclosure.

[00292] As used herein, the term “MIC” means maximal information coefficient. MIC is a type of nonparamentric network analysis that identifies a score (MIC score) between active microbial strains of the present disclosure and at least one measured metadata (e.g., milk fat). Further, U.S. Application No. 15 / 217,575, filed on July 22, 2016 (issued as U.S. Patent No. 9,540,676 on January 10, 2017) is hereby incorporated by reference in its entirety.

[00293] The maximal information coefficient (MIC) is then calculated between strains and metadata 3021a, and between strains 3021b; as seen in FIG. 2. Results are pooled to create a list of all relationships and their corresponding MIC scores 3022. If the relationship scores below a given threshold 3023, the relationship is deemed / identified as irrelevant 3023b. If the relationship is above a given threshold 3023, the relationship deemed / identified as relevant 2023a, and is further subject to network analysis 3024. The following code fragment shows an exemplar}'- methodology for such analysis, according to one embodiment: Read total list of relationships file as links threshold = 0.8 for i in range(len(links)): if links >= threshold multiplier[i] = 1 else multiplier[i] = 0 end if links temp = multiplier*links final links = links temp[links temp!=0] 2026204768   19 Jun 2026 savetxt(output file,final links) output _file. closeQ

[00294] In some embodiments, the compositions of the present disclosure comprise one or more bacteria and / or one or more fungi that have a MIC score of at least about 0.1, at least about 0.15, at least about 0.2, at least about 0.25, at least about 0.3, at least about 0.35, at least about 0.4, at least about 0.45, at least about 0.5, at least about 0.55, at least about 0.6, at least about 0.65, at least about 0.7, at least about 0.75, at least about 0.80, at least about 0.85, at least about 0.9, or at least about 0.95.

[00295] In some embodiments, the compositions of the present disclosure comprise one or more bacteria and / or one or more fungi that have a MIC score of at least 0.1, at least 0.15, at least 0.2, at least 0.25, at least 0.3, at least 0.35, at least 0.4, at least 0.45, at least 0.5, at least 0.55, at least 0.6, at least 0.65, at least 0.7, at least 0.75, at least 0.80, at least 0.85, at least 0.9, or at least 0.95.

[00296] Based on the output of the network analysis, active strains are selected 3025 for preparing products (e.g., ensembles, aggregates, and / or other synthetic groupings) containing the selected strains. The output of the network analysis can also be used to inform the selection of strains for further product composition testing.

[00297] The use of thresholds is discussed above for analyses and determinations. Thresholds can be, depending on the implementation and application: (1) empirically determined (e g., based on distribution levels, setting a cutoff at a number that removes a specified or significant portion of low level reads); (2) any non-zero value; (3) percentage / percentile based; (4) only strains whose normalized second marker (i.e., activity7) reads is greater than normalized first marker (cell count) reads; (5) log2 fold change between activity and quantity or cell count; (6) normalized second marker (activity) reads is greater than mean second marker (activity) reads for entire sample (and / or sample set); and / or any magnitude threshold described above in addition to a statistical threshold (i.e., significance testing). The following example provides thresholding detail for distributions of RNA-based second marker measurements with respect to DNA-based first marker measurements, according to one embodiment. 2026204768   19 Jun 2026

[00298] .As used herein “shelf-stable” refers to a functional attribute and new utility acquired by the microbes formulated according to the disclosure, which enable said microbes to exist in a useful / active state outside of their natural environment in the rumen (i.e. a markedly different characteristic). Thus, shelf-stable is a functional attribute created by the formulations / compositions of the disclosure and denoting that the microbe formulated into a shelf-stable composition can exist outside the rumen and under ambient conditions for a period of time that can be determined depending upon the particular formulation utilized, but in general means that the microbes can be formulated to exist in a composition that is stable under ambient conditions for at least a few days and generally at least one week. Accordingly, a “shelf-stable ruminant supplement” is a composition comprising one or more microbes of the disclosure, said microbes formulated in a composition, such that the composition is stable under ambient conditions for at least one week, meaning that the microbes comprised in the composition (e.g. whole cell, spore, or lysed cell) are able to impart one or more beneficial phenotypic properties to a ruminant when administered (e.g. increased milk yield, improved milk compositional characteristics, improved rumen health, and / or modulation of the rumen microbiome). Feedlot Cattie vs. Dairy Cows

[00299] The instant subject matter is distinct over the subject matter of prior Ascus Biosciences, Inc. applications. The 16S sequences of the microbes of the instant disclosure are believed to be distinct over those of any prior Ascus Biosciences, Inc. applications. One of ordinary skill in the art would be aware that the diet of a dairy' cow would be distinct from that of a steer on a beef feedlot. The steer on the beef feedlot would be fed a high-energy high-gram diet in order to quickly increase the rate of weight gam and to increase the maximum weight prior to rendering. The cow on the daily farm would be fed a different diet that is optimized for the production of milk with little consideration for rapid weight gain or highest maximum weight. The two diets would result in the rumen of the animals in the two environments to yield drastically different microbiota. Thus, the microorganisms in the rumen of the dairy cow and that of the feedlot steer are expected to be different from one another. Isolated Microbes 2026204768   19 Jun 2026

[00300] In some aspects, the present disclosure provides isolated microbes, including novel strains of microbes, presented in Table 1 and Table 2.

[00301] In other aspects, the present disclosure provides isolated whole microbial cultures of the microbes identified in Table 1 and Table 2. These cultures may comprise microbes at various concentrations.

[00302] In some aspects, the disclosure provides for utilizing one or more microbes selected from Table 1 and Table 2 to increase a phenotypic trait of interest in beef cattle.

[0192] In some embodiments, the disclosure provides isolated microbial species belonging to taxonomic families of Prevotellaceae, Veillonellaceae, Ruminococcaceae, Fibrobacteraceae, Bacillaceae 1, Spirochaetaceae, Bacteroidaceae, Lachnospiraceae, Porphyromonadaceae, Coriobacteriaceae, Acidaminococcaceae, Clostridiaceae 1, Rhodobacteraceae, Cryomorphaceae, Erysipelotrichaceae, Promicromonosporaceae, Burkholderiaceae, Succinivibrionaceae, Pseudomonadaceae, Corynebacteriaceae, Planococcaceae, Streptomycetaceae, Synergistaceae, Nocardiopsaceae, Flavobacteriaceae, Propionibactenaceae, Staphylococcaceae, Clostridiales incertae sedis XIII, Anaeroplasmataceae, Pasteurellaceae, Caulobacteraceae, and Sphingomonadaceae.

[0193] In further embodiments, isolated microbial species may be selected from genera of Fibrobacter, Saccharofermentans, Bacillus, Spirochaeta, Bacteroides, Lachnospiracea incertae sedis, Clostridium XLVa, Ruminococcus, Butyricimonas, Olsenella, Acidaminococcus, Parabacteroides, Clostridum sensu stricto, Oribacterium, Pseudoflavonifractor, Treponema, Rhodobacter, Fluviicola, Succiniclasticum, Solobacterium, Veillonella, Cellulosimicrobium, Cupriavidus, Megasphaera, Succinivibrio, Oscillibacter, Pseudomonas, Corynebacterium, Adlercreutzia, Dorea, Roseburia, Anaerovibrio, Sporosarcina, Streptomyces, Syntrophococcus, Butyrivibrio, Lachnobacterium, Pyramidobacter, Coprococcus, Ruminobacter, Thermobifidia, Papillibacter, Aquimarina, Propioniciclava, Staphylococcus, Mogibacterium, Pseudobutyrivibrio, Asteroleplasma, Turicibacter, Aggregatibacter, Brevundimonas, Phascolarctobacterium, and Sphingobium.

[0194] Furthermore, the disclosure relates to microbes having characteristics substantially similar to that of a microbe identified in Table 1 and / or Table 2. 2026204768   19 Jun 2026

[0195] The isolated microbial species, and novel strains of said species, identified in the present disclosure, are able to impart beneficial properties or traits to beef cattle production.

[0196] For instance, the isolated microbes described in Table 1 and Table 2, or microbial ensemble of said microbes, are able to increase feed efficiency. The increase can be quantitatively measured, for example, by measuring the effect that said microbial application has upon the modulation of feed efficiency.

[0197] In some embodiments, the isolated microbial strains are microbes of the present disclosure that have been genetically modified. In some embodiments, the genetically modified or recombinant microbes comprise polynucleotide sequences which do not naturally occur in said microbes. In some embodiments, the microbes may comprise heterologous polynucleotides. In further embodiments, the heterologous polynucleotides may be operably linked to one or more polynucleotides native to the microbes.

[0198] In some embodiments, the heterologous polynucleotides may be reporter genes or selectable markers. In some embodiments, reporter genes may be selected from any of the family of fluorescence proteins (e.g., GFP, RFP, YFP, and the like), P-galactosidase, luciferase. In some embodiments, selectable markers may be selected from neomycin phosphotransferase, hygromycin phosphotransferase, aminoglycoside adenyltransferase, dihydrofolate reductase, acetolactase synthase, bromoxynil nitrilase, P~glucuronidase, dihydrogolate reductase, and chloramphenicol acetyltransferase. In some embodiments, the heterologous polynucleotide may be operably linked to one or more promoter.

[0199] In some embodiments the isolated microbial strains express transgenic or native polypeptides selected from cellulases (endocellulases, exocellulases, glucosidases), pectinases, amylases, amylopectinases, ligninases, and phytases.

[0200] The isolated microbes of Table 2 represent variants of the microbes recited in Table 1. The microbes of Table 2, comprise the reference strains, and the microbes of Table 1 comprise the variants thereof. Generally, the variants of Table 2 comprise 16S rRNA sequences that share at least about 97% sequence identity with the 16S rRNA of the corresponding reference strain in Table B. For example, strains Asbusbbf_873A to Asbusbbf_873G (Table 2 correspond to variants of the reference strain Ascusbbf_873 (Table 1), wherein the 16S rRNA of the variants share at least about 97% sequence identity with that of the reference strain. 2026204768   19 Jun 2026 Microbial Compositions

[0201] In some aspects, the disclosure provides microbial compositions comprising a combination of at least any two microbes selected from amongst the microbes identified in Table 1 and Table 2.

[0202] In certain embodiments, the compositions of the present disclosure comprise two microbes, or three microbes, or four microbes, or five microbes, or six microbes, or seven microbes, or eight microbes, or nine microbes, or ten or more microbes. Said microbes of the compositions are different microbial species, or different strains of a microbial species.

[0203] In some embodiments, the disclosure provides microbial compositions, comprising: at least one or at least two isolated microbial species belonging to genera of: Fibrobacter, Saccharofermentans, Bacillus, Spirochaeta, Bacteroides, Lachnospiracea incertae sedis, Clostridium XLVa, Ruminococcus, Butyricimonas, Olsenella, Acidaminococcus, Parabacteroides, Clostridum sensu stricto, Oribacterium, P seudoflavonifractor, Treponema, Rhodobacter, Fluviicola, Succiniclasticum, Solobacterium, Veillonella, Cellulosimicrobium, Cupriavidus, Megasphaera, Succinivibrio, Oscillibacter, Pseudomonas, Corynebacterium, Adlercreutzia, Dorea, Roseburia, Anaerovibrio, Sporosarcina, Streptomyces, Syntrophococcus, Butyrivibrio, Lachnobacterium, Pyramidobacter, Coprococcus, Ruminobacter, Thennobifidia, Papillibacter, Aquimarina, Propioniciclava, Staphylococcus, Mogibacterium, Pseudobutyrivibrio, Asteroleplasma, Turicibacter, Aggregatibacter, Brevundimonas, Phascolarctobacterium, and Sphingobium. Particular novel strains of species of these aforementioned genera can be found in Table 1 and Table 2.

[0204] In some embodiments, the disclosure provides microbial compositions, comprising: at least one or at least two isolated microbial species belonging to the family of: Prevotellaceae, Veillonellaceae, Ruminococcaceae, Fibrobacteraceae, Bacillaceae 1, Spirochaetaceae, Bacteroidaceae, Lachnospiraceae, Porphyromonadaceae, Coriobacteriaceae, Acidaminococcaceae, Clostridiaceae 1, Rhodobacteraceae, Cryomorphaceae, Erysipelotrichaceae, Promicromonosporaceae, Burkholderiaceae, Succinivibrionaceae, Pseudomonadaceae, Corynebacteriaceae, Planococcaceae, Streptomycetaceae, Synergistaceae, Nocardiopsaceae, Flavobacteriaceae, Propionibacteriaceae, Staphylococcaceae, Clostridiales 2026204768   19 Jun 2026 incertae sedis XIII, Anaeroplasmataceae, Pasteurellaceae, Caulobacteraceae, and Sphingomonadaceae.

[0205] Particular novel strains of species of these aforementioned genera can be found in Table 1 and Table 2.

[0206] In particular aspects, the disclosure provides microbial compositions, comprising species as grouped in Tables 3-9. With respect to Tables 3-9, the letters A through I represent a nonlimiting selection of microbes of the present disclosure, defined as:

[0207] A == Strain designation Ascusbbf 154 identified in Table 1;

[0208] B = Strain designation Ascusbbf 4 identified in Table 1;

[0209] C == Strain designation .Ascusbbf 14146 identified in Table 1;

[0210] D = Strain designation Ascusbbf 876 identified in Table 1;

[0211] E === Strain designation Ascusbbf_24302 identified in Table 1;

[0212] F === Strain designation Ascusbbf J 085 identified in Table 1;

[0213] G == Strain designation Ascusbbf_l identified in Table 1;

[0214] H === Strain designation Ascusbbf 6 r76 identified in Table 1; and

[0215] I === Strain designation Ascusbbf_3427 identified in Table 1. Table 3: Eight and Nine Strain Compositions A,B,C,D,E,F,G,H A,B,C,D,E,F,G,i A.B,C,D,E,F,H,i A,B,C,D,E,G,H,i A,B,C,D,F,G,H,I A,B,C,E,F,G,H,I A,B,D,E,F,G,H,I A,C,D,E,F,G,HJ B,C,D,E,F,G,H,I A,B,C,D,E,F,G,H,I Table 4: Seven Strain Compositions A,B,C,D,E,F,G A,B,C,D,E,F,H A,B,C,D,E,F,I A,B,C,D,E,G,H A,B,C,D,E,G,I A,B,C,D,E,H,I A,B,C,D,F,G,H A,B,C,D,F,G,I A,B,C,D,F,H,I A,B,C,D,G,HJ A,B,C,E,F,G,H A,B,C,E,F,G,I A,B,C,E,F,H,I A,B,C,E,G,H,I A,B,C,F,G,H,I A,B,D,E,F,G,H A,B,D,E,F,G,I a,b,d,e,f,h,i A,B,D,E,G,HJ A,B,D,F,G,H,I A,B,E,F,G,H,I A,C,D,E,F,G,H A,C,D,E,F,GJ A,C,D,E,F,H,i A,C,D,E,G,H,I A,C,D,F,G,H,I A,C,E,F,G,H,I A,D,E,F,G,H,I B,C,D,E,F,G,H B,C,D,E,F,G,I B,C,D,E,F,H,I B,C,D,E,G,H,I B,C,D,F,G,H,I B,C,E,F,G,H,I B,D,E,F,G,H,I C,D,E,F,G,H,I Table 5: Six Strain Compositions A,B,C,D,E,F A,B,C,D,E,G 1 A,B,C,D,E,H A,B,C,D,E,I A,B,C,D,F,G A,B,C,D,F,H A,B,C,D,F,I 2026204768   19 Jun 2026 A,B,C,D,G,H A,B,C,D,G,I A,B,C,D,H,I A,B,C,E,F,G A,B,C,E,F,H A,B,C,E,FJ A,B,C,E,G,H a,b,c,e,g,i A,B,C,E,H,I A,B,C,F,G,H A,B,C,F,G,I a,b,c,f,h,i a,b,c,g,h,i A,B.D,E,F,G a,b,d,e,f,h A,B,D,E,FJ a,b,d,e,g,h a,b,d,e,g,i A,B,D,E,H,I A,B,D,F,G,H A,B,D,F,G,I D,E,F,G,H,i c,e,f,g,h,i a,b,d,f,h,i A,B,D,G,HJ A,B,E,F,G,H A,B,E,F,GJ A,B,E,F,H,I A,B,E,G,HJ A,B,F,G,H,i a,c,d,e,f,g a,c,d,e,f,h a,c,d,e,f,i A,C,D,E,G,H A,C,D,E,G,I A,C,D,E,H,I A,C,D,F,G,H A,C,D,F,GJ a,c,d,f,hj a,c,d,g,h,i A,C,E,F,G,H A,C,E,F,GJ a,c,e,f,h,i a,c,e,g,h,i a,c,f,g,hj a,d,e,f,g,h A,D,E,F,GJ A,D,E,F,H,I A,D,E,G,H,I A,D,F,G,H,I a,e,f,g,h,i B,C,D,E,F,G b,c,d,e,f,h B,C,D,E,F,i B,C,D,E,G,H B,C, D,E,G,i B,C,D,E,H,I B,C,D,F,G,H B,C,D,F,G,I b,c,d,f,h,i B,C,D,G,H,I B,C,E,F,G,H B,C,E,F,G,I B,C,E,F,H,I B,C,E,G,H,I B,C,F,G,H,i b,d,e,f,g,h B,D,E,F,G,I B,D,E,F,H,i B,D,E,G,H,I B,D,F,G,H,I b,e,f,g,h,i C,D,E,F,G,H C,D,E,F,G,I C,D,E,F,H,I C,D,E,G,H,I C,D,F,G,H,i Table 6: Five Strain Compositions a,b,c,d,e a,b,c,d,f a,b,c,d,g A,B,C,D,H A,B,C,D,I A,B,C,E,F a,b,c,e,g A,B,C,E,H A,B,C,F,H A,B,C,F,G A,B..C,F,I A,B,C,G,H A..B,C,GJ A,B,C,H,I A,B,D,E,F a,b,d,e,g A,B,D,E,I A,B,D,F,G a,b,d,f,h A,B,D,FJ a,b,d,g,h A,B,D,G,I a,b,d,h,i a,b,e,f,g A,B,E,F,I A,B,E,G,H a,b,e,g,i A,B,E,HJ A,B,F,G,H A,B,F,G,I A,B,F,H,I A,B,G,H,I A,C,D,E,G a,c,d,e,h A..C,D,E,I A,C,D,F,G a,c,d,f,h A,C,D,F,I A,C.D,G,H A,C,D,G,I A,C,E,F,G a,c,e,f,h A,C,E,F,I A,C,E,G,H A,C,E,G,I A,C,E,HJ A,C,F,G,H a,c,f,gj A,C,G,H,I A,D,E,F,G A,D,E,F,H a,d,e,fj a,d,e,g,h A,D,E,G,I A,D,E,HJ A,D,F,G,H a,d,f,hj A,D,G,HJ A,E,F,G,H A,E,F,G.J A,E,F,H,I A,E,G,H,I A,F,G,H,I b,c,d.e,f B,C,D,E,H B,C,D,E,I B,C,D,F,G B,C,D,F,H B,C?D,F,I B,C,D,G,H b,c,d,g,i b,c.d,hj B,C,E,F,H B,C,E,F,I b,c,e,g,h B,C,E,GJ B,C,E,HJ b,c,f,g,h B,C,F,G,! B,C,F,H,I B,D,E,F,G B,D.E,F,H B,D,E,F,I B,D,E,G,H b,d,e,gj B,D,E,H,I B,D,F,G,H b,d,f,g,i B,D,G,H,I B,E,F,G,H B,E,F,G,I B..E,F,H.I B,E,G,H,I B,F,G,H,I C,D,E,F,G C,D,E,F,H C,D,E,G,H c,d,e,gj c,d,e,h,i C,D,F,G,H QD,F,G,I C,D,F,HJ c,d,g,h,i C,E,F,G,H C,E,F,H,I C,E,G,HJ c,f,g,h,i D,E,F,G,H d,e,f,gj D,E,F,H,I d,e,g,h,i D,F,G,H.I A,B,C,E,I a,b,d,e,h A,B..E,F..H A,C,D..E..F a..c,d,hj A,C,F,H,I a,d,f,g,i B,C,D,E,G b,c,e,f,g B,C,G,HJ B,D;F;H,I QD.E.F.I C,E,F,G,I E,F;G;HJ Tabk 7: Four Strain Compositions A,B,C,D A,B,C,E A;B,C,F A,B,C,G A,B,QH A,B,CJ A,B,D,E a,b,d,f d,g,h,i a,b,d,g A,B,D,H A,B,DJ A,B,E,F A,B,E,G a,b,e,h A,B,E,i a,b,f,g e,f,g,h a,b,f,h a,d,f,h A,D,F,I a,d,g,h a,d,g,i a,d,h,i A,E,F,G A,E,F,H E,F,G,I A,B,F,I A,B,G,H a,b,g,i A,B,HJ A..C,D,E A,C.D,F A,C,D,G a,c,d,h e,f,h,i A,QDJ A,C.E,F a,c,e;g a,c,e,h A,C,E,I a,c,f,g A,C,F,H A;C,FJ E,G,H,I A,C,G,H A,C,G,I A,C,H,I A,D,E,F A,D,E,G A,D,E,H A,D,E,I A,D,F,G F,G,H,i a,e,fj a,e,g,h A,E,GJ a,e,hj a,f,g,h A,F..GJ A,F,H,I a,g,h,i D,E,F,H b,c,d,e B,CfD,F B,QD.G b,c,d,h B,QE,F b,c,e,g B,C,E,H d,e,f,i B,C,EJ B,C,F,G B,C,F,H B,C,FJ B,C,G,H B,C,G,I B,C,H,I b,d,e,f d,e,g,h 2026204768   19 Jun 2026 B,D,E,G B,D,E,H B,D,E,I B,D,F,G B,D,F,H B,D,F,I B,D,G,H B,D,G,I D,E,GJ B,D,HJ CD “H B,E,F,H B,E,FJ B,E,G,H B,E,G,I B,E,HJ B,F,G,H D,E,HJ B,F,GJ B,G,H,I C,D,E,F C,D,E,G C,D,E,H C,D,E,I C,D,F,G D,F,G,H C,D,F,H C,D,F,I C,D,G,H C,D,G,I C,D,HJ C,E,F,G C,E,F,H C,E,F,I D,F,GJ C,E,G,H C.E,G.I C,E,H,I C,F,G,H C,F,GJ C,F,HJ C,G,H,I D,E,F,G D,F,HJ Table 8: Three Strain Compositions A,B,C A,B,D A,B,E A,B,F a,b,g A,B,H A,BJ A,C,D A,C,E G,H,i E,F,H A,C,F A,C,G A,C,H A,C,I A,D,E A,D,F A,D,G A,D,H A,DJ F,H,i E,F,G A,E,F A,E,G A,E,H A,E,I A,F,G A,F,H A,FJ A,G,H A,GJ F;GJ D,H,I A,H,I B,C,D B,C,E B,C,F B,C,G B,C,H B,C,I B,D,E B,D,F F,G,H D,G,I B,D,G B,D,H B,D,I B,E,F B,E,G B,E,H B.EJ B,F,G B,F,H E..H,I E,F,i B,FJ B,G,H B,GJ B,H,I C,D,E C,D,F C,D,G C,D,H C,DJ E,G,I D,G,H C,E,F C,E,G C,E,H C,E,I C,F,G C,F,H C,F,i C,G,H C,G,I E,G,H D,FJ C,HJ D,E,F D,E,G D,E,H D,EJ D,F,G D,F,H Table 9: Two Strain Compositions A,B A,C A,D A,E A,F A,G A,H A J B,C B,D B,E B,F In B,H BJ C,D C,E C,F C,G C,H CJ D,E D,F D,G D,H DJ E,F E,G E,H EJ F,G F,H FJ G,H GJ HJ |

[0216] In some embodiments, the microbial compositions may be selected from any member group from TabHes 3-9. Isolated Microbes - Source Material

[0217] In particular embodiments, the microbes of the present compositions are not naturally found m association with the same animal. In some aspects, the microbial species forming the microbial community are all found in association with animals from the same geographic location. In other aspects, each microbial species forming the composition is from a different geographic location. A geographic location can be defined based upon the predominant soil type in a region, the predominant climate in a region, the predominant plant community present in a region, the predominant plant community present in a region, the distance between regions, the average rainfall in a region, among others. [021§] In some embodiments, the microbes of the present compositions are not naturally found in association with the same species of animal. In some embodiments, the microbes of the 2026204768   19 Jun 2026 present compositions are found in the same species of animal, but separated by geographic region.

[0219] In a particular embodiment, at least one microbial species that is a member of the microbial community derived by the disclosed method is native to, or was acquired from, a geographic region at least about 1 m, 10 m, 100 m, 1 km, 10 km, 100 km, 1,000 km, 10,000 km, 20,000 km, 30,000 km, or 40,000 km from the location of the plant upon which a phenotypic trait is to be increased based upon the taught methods.

[0220] The microbes of the present disclosure were obtained, among other places, at various locales in the United States from the gastrointestinal tract of beef cattle. Isolated Microbes - Microbial Culture Techniques [ 0221] The microbes of Table 1 and Table 2 were matched to their nearest taxonomic groups by utilizing classification tools of the Ribosomal Database Project (RDP) for 16s rRNA sequences and the User-friendly Nordic ITS Ectomycorrhiza (UNITE) database for ITS rRNA sequences. Examples of matching microbes to their nearest taxa may be found in Lan et al. (2012. PLOS one. 7(3):e32491), Schloss and Westcott (2011. Appl. Environ. Microbiol. 77(10):3219-3226), andKoljalg et al. (2005. New Phytologist. 166(3):1063-1068).

[0222] The isolation, identification, and culturing of the microbes of the present disclosure can be effected using standard microbiological techniques. Examples of such techniques may be found in Gerhardt, P. (ed.) Methods for General and Molecular Microbiology. American Society for Microbiology, Washington, D.C. (1994) and Lennette, E. H. (ed.) Manual of Clinical Microbiology, Third Edition. American Society' for Microbiology, Washington, D.C. (1980), each of which is incorporated by reference.

[0223] Isolation can be effected by streaking the specimen on a solid medium (e.g., nutrient agar plates) to obtain a single colony, which is characterized by the phenotypic traits described hereinabove (e.g., Gram positive / negative, capable of forming spores aerobically / anaerobically, cellular morphology, carbon source metabolism, acid / base production, enzyme secretion, metabolic secretions, etc.) and to reduce the likelihood of working with a culture which has become contaminated. 2026204768   19 Jun 2026

[0224] For example, for microbes of the disclosure, biologically pure isolates can be obtained through repeated subculture of biological samples, each subculture followed by streaking onto solid media to obtain individual colonies or colony forming units. Methods of preparing, thawing, and growing lyophilized bacteria are commonly known, for example, Gherna, R. L. and C. A. Reddy. 2007. Culture Preservation, p 1019-1033. In C. A. Reddy, T. J. Beveridge, J. A. Breznak, G. A. Marzluf, T. M. Schmidt, and L. R. Snyder, eds. American Society for Microbiology, Washington, DC., 1033 pages; herein incorporated by reference. Thus freeze dried liquid formulations and cultures stored long term at ”70° C in solutions containing glycerol are contemplated for use in providing formulations of the present disclosure.

[0225] The microbes of the disclosure can be propagated in a liquid medium under aerobic conditions, or alternatively anaerobic conditions. Medium for growing the bacterial strains of the present disclosure includes a carbon source, a nitrogen source, and inorganic salts, as well as specially required substances such as vitamins, amino acids, nucleic acids and the like. Examples of suitable carbon sources which can be used for growing the microbes include, but are not limited to, starch, peptone, yeast extract, amino acids, sugars such as glucose, arabinose, mannose, glucosamine, maltose, and the like; salts of organic acids such as acetic acid, fumaric acid, adipic acid, propionic acid, citric acid, gluconic acid, malic acid, pyruvic acid, malonic acid and the like; alcohols such as ethanol and glycerol and the like; oil or fat such as soybean oil, rice bran oil, olive oil, corn oil, sesame oil. The amount of the carbon source added varies according to the kind of carbon source and is typically between 1 to 100 gram(s) per liter of medium. Preferably, glucose, starch, and / or peptone is contained in the medium as a major carbon source, at a concentration of 0.1-5% (W / V). Examples of suitable nitrogen sources which can be used for growing the bacterial strains of the present disclosure include, but are not limited to, ammo acids, yeast extract, tryptone, beef extract, peptone, potassium nitrate, ammonium nitrate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonia or combinations thereof. The amount of nitrogen source varies according to the type of nitrogen source, typically between 0.1 to 30 gram per liter of medium. The inorganic salts, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, magnesium sulfate, magnesium chloride, ferric sulfate, ferrous sulfate, ferric chloride, ferrous chloride, manganous sulfate, manganous chloride, zinc sulfate, zinc chloride, cupric sulfate, calcium chloride, sodium chloride, calcium carbonate, sodium carbonate can be used alone or in 2026204768   19 Jun 2026 combination. The amount of inorganic acid varies according to the kind of the inorganic salt, typically between 0.001 to 10 gram per liter of medium. Examples of specially required substances include, but are not limited to, vitamins, nucleic acids, yeast extract, peptone, meat extract, malt extract, dried yeast and combinations thereof. Cultivation can be effected at a temperature, which allows the growth of the microbial strains, essentially, between 20°C and 46°C. In some aspects, a temperature range is 30°C-39°C. For optimal growth, in some embodiments, the medium can be adjusted to pH 6.0-7.4. It will be appreciated that commercially available media may also be used to culture the microbial strains, such as Nutrient Broth or Nutrient Agar available from Difco, Detroit, MI. It will be appreciated that cultivation time may differ depending on the type of culture medium used and the concentration of sugar as a major carbon source.

[0226] In some aspects, cultivation lasts between 24-96 hours. Microbial cells thus obtained are isolated using methods, which are well known in the art. Examples include, but are not limited to, membrane filtration and centrifugal separation. The pH may be adjusted using sodium hydroxide and the like and the culture may be dried using a freeze dryer, until the water content becomes equal to 4% or less. Microbial co-cultures may be obtained by propagating each strain as described hereinabove. In some aspects, microbial multi-strain cultures may be obtained by propagating two or more of the strains described hereinabove. It will be appreciated that the microbial strains may be cultured together when compatible culture conditions can be employed. Isolated Microbes - Microbial Strains

[0227] Microbes can be distinguished into a genus based on polyphasic taxonomy, which incorporates all available phenotypic and genotypic data into a consensus classification (Vandamme et al. 1996. Polyphasic taxonomy, a consensus approach to bacterial systematics. Microbiol Rev 1996, 60:407-438). One accepted genotypic method for defining species is based on overall genomic relatedness, such that strains which share approximately 70% or more relatedness using DNA-DNA hybridization, with 5°C or less A7A(the difference in the melting temperature between homologous and heterologous hybrids), under standard conditions, are considered to be members of the same species. Thus, populations that share greater than the aforementioned 70% threshold can be considered to be variants of the same species. Another accepted genotypic method for defining species is to isolate marker genes of the present 2026204768   19 Jun 2026 disclosure, sequence these genes, and align these sequenced genes from multiple isolates or variants. The microbes are interpreted as belonging to the same species if one or more of the sequenced genes share at least 97% sequence identity.

[0228] The 16S or 18S rRNA sequences or ITS sequences are often used for making distinctions between species and strains, in that if one of the aforementioned sequences shares less than a specified % sequence identity from a reference sequence, then the two organisms from which the sequences were obtained are said to be of different species or strains.

[0229] Thus, one could consider microbes to be of the same species, if they share at least 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identify across the 16S or 18S rRNA sequence, or the ITS1 or ITS2 sequence.

[0230] Further, one could define microbial strains of a species, as those that share at least 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identify' across the 16S or 18S rRNA sequence, or the ITS1 or ITS2 sequence.

[0231] In one embodiment, microbial strains of the present disclosure include those that comprise polynucleotide sequences that share at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with any one of SEQ ID NOs:l-5,993. In a further embodiment, microbial strains of the present disclosure include those that comprise polynucleotide sequences that share at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identify with any one of SEQ ID NOs: 1-5,993.

[0232] Comparisons may also be made with 23 S rRNA sequences against reference sequences.

[0233] Unculturable microbes often cannot be assigned to a definite species in the absence of a phenotype determination, the microbes can be given a candidatus designation within a genus provided their 16S or 18S rRNA sequences or ITS sequences subscribes to the principles of identity with known species.

[0234] One approach is to observe the distribution of a large number of strains of closely related species in sequence space and to identify clusters of strains that are well resolved from other clusters. This approach has been developed by using the concatenated sequences of multiple core 2026204768   19 Jun 2026 (house-keeping) genes to assess clustering patterns, and has been called multilocus sequence analysis (MLSA) or multilocus sequence phylogenetic analysis. MLSA has been used successfully to explore clustering patterns among large numbers of strains assigned to very closely related species by current taxonomic methods, to look at the relationships between small numbers of strains within a genus, or within a broader taxonomic grouping, and to address specific taxonomic questions. More generally, the method can be used to ask whether bacterial species exist - that is, to observe whether large populations of similar strains invariably fall into well-resolved clusters, or whether in some cases there is a genetic continuum in which clear separation into clusters is not observed.

[0235] In order to more accurately make a determination of genera, a determination of phenotypic traits, such as morphological, biochemical, and physiological characteristics are made for comparison with a reference genus archetype. The colony morphology can include color, shape, pigmentation, production of slime, etc. Features of the cell are described as to shape, size, Gram reaction, extracellular material, presence of endospores, flagella presence and location, motility, and inclusion bodies. Biochemical and physiological features describe growth of the organism at different ranges of temperature, pH, salinity and atmospheric conditions, growth in presence of different sole carbon and nitrogen sources. One of ordinary skill in the art would be reasonably apprised as to the phenotypic traits that define the genera of the present disclosure.

[0236] In one embodiment, the microbes taught herein were identified utilizing 16S rRNA gene sequences and ITS sequences. It is known in the art that 16S rRNA contains hypervariable regions that can provide species / strain-specific signature sequences useful for bacterial identification, and that ITS sequences can also provide species / strain-specific signature sequences useful for fungal identification.

[0237] Phylogenetic analysis using the rRNA genes and / or ITS sequences are used to define “substantially similar” species belonging to common genera and also to define “substantially similar” strains of a given taxonomic species. Furthermore, physiological and / or biochemical properties of the isolates can be utilized to highlight both minor and significant differences between strains that could lead to advantageous behavior in beef cattle. 2026204768   19 Jun 2026

[0238] Compositions of the present disclosure may include combinations of fungal spores and bacterial spores, fungal spores and bacterial vegetative cells, fungal vegetative cells and bacterial spores, fungal vegetative cells and bacterial vegetative cells. In some embodiments, compositions of the present disclosure comprise bacteria only in the form of spores. In some embodiments, compositions of the present disclosure comprise bacteria only in the form of vegetative cells. In some embodiments, compositions of the present disclosure comprise bacteria in the absence of fungi. In some embodiments, compositions of the present disclosure comprise fungi in the absence of bacteria. In some embodiments, compositions of the present disclosure comprise VBNC bacteria and / or fungi. In some embodiments, compositions of the present disclosure comprise bacteria and / or fungi in a quiescent state. In some embodiments, compositions of the present disclosure include dormant bacteria and / or fungi.

[0239] Bacterial spores may include endospores and akinetes. Fungal spores may include statismospores, ballistospores, autospores, aplanospores, zoospores, mitospores, megaspores, microspores, meiospores, chlamydospores, uredmiospores, teliospores, oospores, carpospores, tetraspores, sporangiospores, zygospores, ascospores, basidiospores, ascospores, and asciospores.

[0240] In some embodiments, spores of the composition germinate upon administration to animals of the present disclosure. In some embodiments, spores of the composition germinate only upon administration to animals of the present disclosure. Microbial Compositions

[0241] In some embodiments, the microbes of the disclosure are combined into microbial compositions.

[0242] In some embodiments, the microbial compositions include cattle feed, such as grain and grain byproducts (barley, maize, oats, sorghum, wheat, distillers grains, sweet bran, and the like); roughage (alfalfa, silage, fescue, clover, ryegrass, and the like); starches (tapioca and the like); protein (oilseed cakes, vegetable wastes, corn by-products, wheat by-products, and the like); liquid feeds (condensed corn distillers solubles, molasses, tallow, yellow grease, corn oil, and the like); or non-nitrogen protein. In some embodiments, the microbial compositions include vitamins and / or metabolites thereof / minerals, urea, trace elements, emulsifiers, aromatizing 2026204768   19 Jun 2026 products, binders, colorants, odorants, thickening agents, antibiotics, and the like. In some embodiments, the microbial compositions include one or more of an ionophore; vaccine, antibiotic; antihelmmtic; virucide; nematicide; amino acids such as methionine, glutamine, valine, glycine, cysteine, homocysteine, aspartic acid, and arginine; fish oil; oregano; carnitine, pantoate, pantothenate, aspartate, and biologically active molecules such as enzymes.

[0243] In some embodiments, the vitamins include vitamin B5, Bl, B2, B3, B6, B9, B12, H, C, A, D, E, or K; and combinations thereof. In some embodiments, the microbial compositions include microbes that synthesize vitamin B5, Bl, B2, B3, B6, B9, B12, H, C, A, D, E, and / or K. In some embodiments, the microbial compositions include microbes that synthesize vitamin B5. In some embodiments, the metabolites of vitamin B5, Bl, B2, B3, B6, B9, B12, H, C, A, D, E, or K are contemplated as one or more components of a microbial composition of the present disclosure. In one embodiment, pantothenate is a component of a microbial composition of the present disclosure. In one embodiment, a component of a microbial composition of the present disclosure includes one or more precursors utilized by mammalian or microbial biosynthesis of vitamins.

[0244] In some embodiments, the microbial compositions of the present disclosure are solid. Where solid compositions are used, it may be desired to include one or more carrier materials including, but not limited to: mineral earths such as silicas, talc, kaolin, limestone, chalk, clay, dolomite, diatomaceous earth; calcium sulfate; magnesium sulfate; magnesium oxide; zeolites, calcium carbonate; magnesium carbonate; trehalose; chitosan; shellac; albumins; starch; skim milk powder; sweet whey powder; maltodextrin; lactose; inulin; dextrose; and products of vegetable origin such as cereal meals, tree bark meal, wood meal, and nutshell meal.

[0245] In some embodiments, the microbial compositions of the present disclosure are liquid. In further embodiments, the liquid comprises a solvent that may include water or an alcohol or a saline or carbohydrate solution, and other animal-safe solvents. In some embodiments, the microbial compositions of the present disclosure include binders such as animal-safe polymers, carboxymethylcellulose, starch, polyvinyl alcohol, and the like.

[0246] In some embodiments, the microbial compositions of the present disclosure comprise thickening agents such as silica, clay, natural extracts of seeds or seaweed, synthetic derivatives of cellulose, guar gum, locust bean gum, alginates, and methylcelluloses. In some embodiments, 2026204768   19 Jun 2026 the microbial compositions comprise anti-settlmg agents such as modified starches, polyvinyl alcohol, xanthan gum, and the like.

[0247] In some embodiments, the microbial compositions of the present disclosure comprise colorants including organic chromophores classified as nitroso; nitro; azo, including monoazo, bisazo and polyazo; acridine, anthraquinone, azine, diphenylmethane, indamine, indophenol, methine, oxazine, phthalocyanine, thiazine, thiazole, triarylmethane, xanthene. In some embodiments, the microbial compositions of the present disclosure comprise trace nutrients such as salts of iron, manganese, boron, copper, cobalt, molybdenum and zinc. In some embodiments, the microbial compositions comprise dyes, both natural and artificial. In some embodiments, the dye is green in color.

[0248] In some embodiments, the microbial compositions of the present disclosure comprise an animal-safe virucide, parasiticide, bacteriocide, fungicide, or nematicide.

[0249] In some embodiments, microbial compositions of the present disclosure comprise saccharides (e.g., monosaccharides, disacchandes, trisaccharides, polysaccharides, oligosaccharides, and the like), polymeric saccharides, lipids, polymeric lipids, lipopolysaccharides, proteins, polymeric proteins, lipoproteins, nucleic acids, nucleic acid polymers, silica, inorganic salts and combinations thereof. In a further embodiment, microbial compositions comprise polymers of agar, agarose, gelrite, gellan gum, and the like. In some embodiments, microbial compositions comprise plastic capsules, emulsions (e.g., water and oil), membranes, and artificial membranes. In some embodiments, emulsions or linked polymer solutions may comprise microbial compositions of the present disclosure. See Harel and Bennett (US Patent 8,460,726 B2).

[0250] In some embodiments, microbial compositions of the present disclosure comprise one or more exygen scavengers, denitrifies, nitrifiers, heavy metal chelators, and / or dechlorinators; and combinations thereof. In one embodiment, the one or more exygen scavengers, denitrifiers, nitrifiers, heavy metal chelators, and / or dechlorinators are not chemically active once the microbial compositions are mixed with food and / or water to be administered to the animal. In one embodiment, the one or more oxygen scavengers, denitrifiers, nitrifiers, heavy metal chelators, and / or dechlorinators are not chemically active when administered to the animal. 2026204768   19 Jun 2026

[0251] In some embodiments, microbial compositions of the present disclosure occur in a solid form (e.g., dispersed lyophilized spores) or a liquid form (microbes interspersed in a storage medium). In some embodiments, microbial compositions of the present disclosure are added in dry form to a liquid to a liquid to form a suspension immediately prior to administration

[0252] In some embodiments, microbial compositions of the present disclosure comprise one or more preservatives. The preservatives may be in liquid or gas formulations. The preservatives may be selected from one or more of monosaccharide, disaccharide, trisaccharide, polysaccharide, acetic acid, ascorbic acid, calcium ascorbate, erythorbic acid, iso-ascorbic acid, erythrobic acid, potassium nitrate, sodium ascorbate, sodium erythorbate, sodium iso-ascorbate, sodium nitrate, sodium nitrite, nitrogen, benzoic acid, calcium sorbate, ethyl lauroyl arginate, methyl- / ?-hydroxy benzoate, methyl paraben, potassium acetate, potassium benzoiate, potassium bisulphite, potassium diacetate, potassium lactate, potassium metabisulphite, potassium sorbate, propylyvhydroxy benzoate, propyl paraben, sodium acetate, sodium benzoate, sodium bisulphite, sodium nitrite, sodium diacetate, sodium lactate, sodium metabisulphite, sodium salt of methyl-p-hydroxy benzoic acid, sodium salt of propyl-p-hydroxy benzoic acid, sodium sulphate, sodium sulfite, sodium dithionite, sulphurous acid, calcium propionate, dimethyl dicarbonate, natamycin, potassium sorbate, potassium bisulfite, potassium metabisulfite, propionic acid, sodium diacetate, sodium propionate, sodium sorbate, sorbic acid, ascorbic acid, ascorbyl palmitate, ascorbyl stearate, butylated hydro-xyanisole, butylated hydroxytoluene (BUT), butylated hydroxyl anisole (BHA), citric acid, citric acid esters of mono- and / or diglycerides, L-cysteine, L-cysteine hydrochloride, gum guaiacum, gum guaiac, lecithin, lecithin citrate, monoglyceride citrate, monoisopropyl citrate, propyl gallate, sodium metabisulphite, tartaric acid, tertiary butyl hydroquinone, stannous chloride, thiodipropionic acid, dilauryl thiodipropionate, distearyl thiodipropionate, ethoxyquin, sulfur dioxide, formic acid, or tocopherol(s).

[0253] In some embodiments, microbial compositions of the present disclosure comprise one or more oxygen scavengers, denitrifiers, nitrifiers, heavy metal chelators, and / or dechlorinators; and combinations thereof. In one embodiment, the one or more oxygen scavengers, denitrifiers, nitrifiers, heavy metal chelators, and / or dechlorinators are not chemically active once the microbial compositions are mixed with food and / or water to be administered to the beef cattle. In 2026204768   19 Jun 2026 one embodiment, the one or more oxygen scavengers, denitrifiers, nitrifiers, heavy metal chelators, and / or dechlorinators are not chemically active when administered to the beef cattle.

[0254] In some embodiments, microbial compositions of the present disclosure include bacterial and / or fungal cells in spore form, vegetative cell form, and / or lysed cell form. In one embodiment, the lysed cell form acts as a mycotoxin binder, e.g. mycotoxins binding to dead cells.

[0255] In some embodiments, the microbial compositions are shelf stable in a refrigerator (3540°F) for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 days. In some embodiments, the microbial compositions are shelf stable in a refrigerator (35-40°F) for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 weeks.

[0256] In some embodiments, the microbial compositions are shelf stable at room temperature (68-72°F) or between 50-77°F for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, II, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 days. In some embodiments, the microbial compositions are shelf stable at room temperature (68-72°F) or between 50-77°F for a period of at least 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 weeks.

[0257] In some embodiments, the microbial compositions are shelf stable at -23-35°F for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 days. In some embodiments, the microbial compositions are shelf stable at -23-35°F for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 weeks.

[0258] In some embodiments, the microbial compositions are shelf stable at 77-100°F for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 2026204768   19 Jun 2026 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 days. In some embodiments, the microbial compositions are shelf stable at 77-100°F for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 weeks.

[0259] In some embodiments, the microbial compositions are shelf stable at 101-213°F for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 days. In some embodiments, the microbial compositions are shelf stable at 101-213°F for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 weeks.

[0260] In some embodiments, the microbial compositions of the present disclosure are shelf stable at refrigeration temperatures (35-40°F), at room temperature (68-72°F), between 50-77°F, between -23-35°F, between 70-100°F, or between 101-213°F for a period of about 1 to 100, about 1 to 95, about 1 to 90, about 1 to 85, about 1 to 80, about 1 to 75, about 1 to 70, about 1 to 65, about 1 to 60, about 1 to 55, about 1 to 50, about 1 to 45, about 1 to 40, about 1 to 35, about 1 to 30, about 1 to 25, about 1 to 20, about 1 to 15, about 1 to 10, about 1 to 5, about 5 to 100, about 5 to 95, about 5 to 90, about 5 to 85, about 5 to 80, about 5 to 75, about 5 to 70, about 5 to 65, about 5 to 60, about 5 to 55, about 5 to 50, about 5 to 45, about 5 to 40, about 5 to 35, about 5 to 30, about 5 to 25, about 5 to 20, about 5 to 15, about 5 to 10, about 10 to 100, about 10 to 95, about 10 to 90, about 10 to 85, about 10 to 80, about 10 to 75, about 10 to 70, about 10 to 65, about 10 to 60, about 10 to 55, about 10 to 50, about 10 to 45, about 10 to 40, about 10 to 35, about 10 to 30, about 10 to 25, about 10 to 20, about 10 to 15, about 15 to 100, about 15 to 95, about 15 to 90, about 15 to 85, about 15 to 80, about 15 to 75, about 15 to 70, about 15 to 65, about 15 to 60, about 15 to 55, about 15 to 50, about 15 to 45, about 15 to 40, about 15 to 35, about 15 to 30, about 15 to 25, about 15 to 20, about 20 to 100, about 20 to 95, about 20 to 90, about 20 to 85, about 20 to 80, about 20 to 75, about 20 to 70, about 20 to 65, about 20 to 60, about 20 to 55, about 20 to 50, about 20 to 45, about 20 to 40, about 20 to 35, about 20 to 30, about 20 to 25, about 25 to 100, about 25 to 95, about 25 to 90, about 25 to 85, about 25 to 80, about 25 to 75, about 25 to 70, about 25 to 65, about 25 to 60, about 25 to 55, about 25 to 50, 2026204768   19 Jun 2026 about 25 to 45, about 25 to 40, about 25 to 35, about 25 to 30, about 30 to 100, about 30 to 95, about 30 to 90, about 30 to 85, about 30 to 80, about 30 to 75, about 30 to 70, about 30 to 65, about 30 to 60, about 30 to 55, about 30 to 50, about 30 to 45, about 30 to 40, about 30 to 35, about 35 to 100, about 35 to 95, about 35 to 90, about 35 to 85, about 35 to 80, about 35 to 75, about 35 to 70, about 35 to 65, about 35 to 60, about 35 to 55, about 35 to 50, about 35 to 45, about 35 to 40, about 40 to 100, about 40 to 95, about 40 to 90, about 40 to 85, about 40 to 80, about 40 to 75, about 40 to 70, about 40 to 65, about 40 to 60, about 40 to 55, about 40 to 50, about 40 to 45, about 45 to 100, about 45 to 95, about 45 to 90, about 45 to 85, about 45 to 80, about 45 to 75, about 45 to 70, about 45 to 65, about 45 to 60, about 45 to 55, about 45 to 50, about 50 to 100, about 50 to 95, about 50 to 90, about 50 to 85, about 50 to 80, about 50 to 75, about 50 to 70, about 50 to 65, about 50 to 60, about 50 to 55, about 55 to 100, about 55 to 95, about 55 to 90, about 55 to 85, about 55 to 80, about 55 to 75, about 55 to 70, about 55 to 65, about 55 to 60, about 60 to 100, about 60 to 95, about 60 to 90, about 60 to 85, about 60 to 80, about 60 to 75, about 60 to 70, about 60 to 65, about 65 to 100, about 65 to 95, about 65 to 90, about 65 to 85, about 65 to 80, about 65 to 75, about 65 to 70, about 70 to 100, about 70 to 95, about 70 to 90, about 70 to 85, about 70 to 80, about 70 to 75, about 75 to 100, about 75 to 95, about 75 to 90, about 75 to 85, about 75 to 80, about 80 to 100, about 80 to 95, about 80 to 90, about 80 to 85, about 85 to 100, about 85 to 95, about 85 to 90, about 90 to 100, about 90 to 95, or 95 to 100 weeks

[0261] In some embodiments, the microbial compositions of the present disclosure are shelf stable at refrigeration temperatures (35-40°F), at room temperature (68-72°F), between 50-77°F, between -23-35°F, between 70-100°F, or between 101-213°F for a period of 1 to 100, 1 to 95, 1 to 90, 1 to 85, 1 to 80, 1 to 75, 1 to 70, 1 to 65, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 1 to 35, 1 to 30, 1 to 25, 1 to 20, 1 to 15, 1 to 10, 1 to 5, 5 to 100, 5 to 95, 5 to 90, 5 to 85, 5 to 80, 5 to 75, 5 to 70, 5 to 65, 5 to 60, 5 to 55, 5 to 50, 5 to 45, 5 to 40, 5 to 35, 5 to 30, 5 to 25, 5 to 20, 5 to 15, 5 to 10, 10 to 100, 10 to 95, 10 to 90, 10 to 85, 10 to 80, 10 to 75, 10 to 70, 10 to 65, 10 to 60, 10 to 55, 10 to 50, 10 to 45, 10 to 40, 10 to 35, 10 to 30, 10 to 25, 10 to 20, 10 to 15, 15 to 100, 15 to 95, 15 to 90, 15 to 85, 15 to 80, 15 to 75, 15 to 70, 15 to 65, 15 to 60, 15 to 55, 15 to 50, 15 to 45, 15 to 40, 15 to 35, 15 to 30, 15 to 25, 15 to 20, 20 to 100, 20 to 95, 20 to 90, 20 to 85, 20 to 80, 20 to 75, 20 to 70, 20 to 65, 20 to 60, 20 to 55, 20 to 50, 20 to 45, 20 to 40, 20 to 35, 20 to 30, 20 to 25, 25 to 100, 25 to 95, 25 to 90, 25 to 85, 25 to 80, 25 to 75, 25 to 70, 25 to 2026204768   19 Jun 2026 65, 25 to 60, 25 to 55, 25 to 50, 25 to 45, 25 to 40, 25 to 35, 25 to 30, 30 to 100, 30 to 95, 30 to 90, 30 to 85, 30 to 80, 30 to 75, 30 to 70, 30 to 65, 30 to 60, 30 to 55, 30 to 50, 30 to 45, 30 to 40, 30 to 35, 35 to 100, 35 to 95, 35 to 90, 35 to 85, 35 to 80, 35 to 75, 35 to 70, 35 to 65, 35 to 60, 35 to 55, 35 to 50, 35 to 45, 35 to 40, 40 to 100, 40 to 95, 40 to 90, 40 to 85, 40 to 80, 40 to 75, 40 to 70, 40 to 65, 40 to 60, 40 to 55, 40 to 50, 40 to 45, 45 to 100, 45 to 95, 45 to 90, 45 to 85, 45 to 80, 45 to 75, 45 to 70, 45 to 65, 45 to 60, 45 to 55, 45 to 50, 50 to 100, 50 to 95, 50 to 90, 50 to 85, 50 to 80, 50 to 75, 50 to 70, 50 to 65, 50 to 60, 50 to 55, 55 to 100, 55 to 95, 55 to 90, 55 to 85, 55 to 80, 55 to 75, 55 to 70, 55 to 65, 55 to 60, 60 to 100, 60 to 95, 60 to 90, 60 to 85, 60 to 80, 60 to 75, 60 to 70, 60 to 65, 65 to 100, 65 to 95, 65 to 90, 65 to 85, 65 to 80, 65 to 75, 65 to 70, 70 to 100, 70 to 95, 70 to 90, 70 to 85, 70 to 80, 70 to 75, 75 to 100, 75 to 95, 75 to 90, 75 to 85, 75 to 80, 80 to 100, 80 to 95, 80 to 90, 80 to 85, 85 to 100, 85 to 95, 85 to 90, 90 to 100, 90 to 95, or 95 to 100 weeks.

[0262] In some embodiments, the microbial compositions of the present disclosure are shelf stable at refrigeration temperatures (35-40°F), at room temperature (68-72°F), between 50-77°F, between -23-35°F, between 70-100°F, or between 101-213°F for a period of about 1 to 36, about 1 to 34, about 1 to 32, about 1 to 30, about 1 to 28, about 1 to 26, about 1 to 24, about 1 to 22, about 1 to 20, about 1 to 18, about 1 to 16, about 1 to 14, about 1 to 12, about 1 to 10, about 1 to 8, about 1 to 6, about 1 one 4, about 1 to 2, about 4 to 36, about 4 to 34, about 4 to 32, about 4 to 30, about 4 to 28, about 4 to 26, about 4 to 24, about 4 to 22, about 4 to 20, about 4 to 18, about 4 to 16, about 4 to 14, about 4 to 12, about 4 to 10, about 4 to 8, about 4 to 6, about 6 to 36, about 6 to 34, about 6 to 32, about 6 to 30, about 6 to 28, about 6 to 26, about 6 to 24, about 6 to 22, about 6 to 20, about 6 to 18, about 6 to 16, about 6 to 14, about 6 to 12, about 6 to 10, about 6 to 8, about 8 to 36, about 8 to 34, about 8 to 32, about 8 to 30, about 8 to 28, about 8 to 26, about 8 to 24, about 8 to 22, about 8 to 20, about 8 to 18, about 8 to 16, about 8 to 14, about 8 to 12, about 8 to 10, about 10 to 36, about 10 to 34, about 10 to 32, about 10 to 30, about 10 to 28, about 10 to 26, about 10 to 24, about 10 to 22, about 10 to 20, about 10 to 18, about 10 to 16, about 10 to 14, about 10 to 12, about 12 to 36, about 12 to 34, about 12 to 32, about 12 to 30, about 12 to 28, about 12 to 26, about 12 to 24, about 12 to 22, about 12 to 20, about 12 to 18, about 12 to 16, about 12 to 14, about 14 to 36, about 14 to 34, about 14 to 32, about 14 to 30, about 14 to 28, about 14 to 26, about 14 to 24, about 14 to 22, about 14 to 20, about 14 to 18, about 14 to 16, about 16 to 36, about 16 to 34, about 16 to 32, about 16 to 30, about 16 to 28, 2026204768   19 Jun 2026 about 16 to 26, about 16 to 24, about 16 to 22, about 16 to 20, about 16 to 18, about 18 to 36, about 18 to 34, about 18 to 32, about 18 to 30, about 18 to 28, about 18 to 26, about 18 to 24, about 18 to 22, about 18 to 20, about 20 to 36, about 20 to 34, about 20 to 32, about 20 to 30, about 20 to 28, about 20 to 26, about 20 to 24, about 20 to 22, about 22 to 36, about 22 to 34, about 22 to 32, about 22 to 30, about 22 to 28, about 22 to 26, about 22 to 24, about 24 to 36, about 24 to 34, about 24 to 32, about 24 to 30, about 24 to 28, about 24 to 26, about 26 to 36, about 26 to 34, about 26 to 32, about 26 to 30, about 26 to 28, about 28 to 36, about 28 to 34, about 28 to 32, about 28 to 30, about 30 to 36, about 30 to 34, about 30 to 32, about 32 to 36, about 32 to 34, or about 34 to 36 months.

[0263] In some embodiments, the microbial compositions of the present disclosure are shelf stable at refrigeration temperatures (35-40°F), at room temperature (68-72°F), between 50-77°F, between -23-35°F, between 70-100°F, or between 101-213°F for a period of 1 to 36 1 to 34 1 to 32 1 to 30 1 to 28 1 to 26 1 to 24 1 to 22 1 to 20 1 to 18 1 to 16 1 to 14 1 to 12 1 to 10 1 to 8 1 to 6 1 one 4 1 to 2 4 to 36 4 to 34 4 to 32 4 to 30 4 to 28 4 to 26 4 to 24 4 to 22 4 to 20 4 to 18 4 to 16 4 to 14 4 to 12 4 to 10 4 to 8 4 to 6 6 to 36 6 to 34 6 to 32 6 to 30 6 to 28 6 to 26 6 to 24 6 to 22 6 to 20 6 to 18 6 to 16 6 to 14 6 to 12 6 to 10 6 to 8 8 to 36 8 to 34 8 to 32 8 to 30 8 to 28 8 to 26 8 to 24 8 to 22 8 to 20 8 to 18 8 to 16 8 to 14 8 to 12 8 to 10 10 to 36 10 to 34 10 to 32 10 to 30 10 to 28 10 to 26 10 to 24 10 to 22 10 to 20 10 to 18 10 to 16 10 to 14 10 to 12 12 to 36 12 to 34 12 to 32 12 to 30 12 to 28 12 to 26 12 to 24 12 to 22 12 to 20 12 to 18 12 to 16 12 to 14 14 to 36 14 to 34 14 to 32 14 to 30 14 to 28 14 to 26 14 to 24 14 to 22 14 to 20 14 to 18 14 to 16 16 to 36 16to34 16 to 32 16to30 16to28 16to26 16to24 16to22 16 to 20 16 to 18 18to36 18to 34 18 to 32 18 to 30 18 to 28 18 to 26 1 8 to 24 18 to 22 18 to 20 20 to 36 20 to 34 20 to 32 20 to 30 20 to 28 20 to 26 20 to 24 20 to 22 22 to 36 22 to 34 22 to 32 22 to 30 22 to 28 22 to 26 22 to 24 24 to 36 24 to 34 24 to 32 24 to 30 24 to 28 24 to 26 26 to 36 26 to 34 26 to 32 26 to 30 26 to 28 28 to 36 28 to 34 28 to 32 28 to 30 30 to 36 30 to 34 30 to 32 32 to 36 32 to 34, or about 34 to 36.

[0264] In some embodiments, the microbial compositions of the present disclosure are shelf stable at any of the disclosed temperatures and / or temperature ranges and spans of time at a relative humidity of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 2026204768   19 Jun 2026 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, or 98%.

[0265] In some embodiments, the microbial composition of the present disclosure possesses a water activity (aw) of less than 0.750, 0.700, 0.650, 0.600, 0.550, 0.500, 0.475, 0.450, 0.425, 0.400, 0.375, 0.350, 0.325, 0.300, 0.275, 0.250, 0.225, 0.200, 0.190, 0.180, 0.170, 0.160, 0.150, 0.140, 0.130, 0.120, 0.110, 0.100, 0.095, 0.090, 0.085, 0.080, 0.075, 0.070, 0.065, 0.060, 0.055, 0.050, 0.045, 0.040, 0.035, 0.030, 0.025, 0.020, 0.015, 0.010, or 0.005.

[0266] In some embodiments, the microbial composition of the present disclosure possesses a water activity (aw) of less than about 0.750, about 0.700, about 0.650, about 0.600, about 0.550, about 0.500, about 0.475, about 0.450, about 0.425, about 0.400, about 0.375, about 0.350, about 0.325, about 0.300, about 0.275, about 0.250, about 0.225, about 0.200, about 0.190, about 0.180, about 0.170, about 0.160, about 0.150, about 0.140, about 0.130, about 0.120, about 0.110, about 0.100, about 0.095, about 0.090, about 0.085, about 0.080, about 0.075, about 0.070, about 0.065, about 0.060, about 0.055, about 0.050, about 0.045, about 0.040, about 0.035, about 0.030, about 0.025, about 0.020, about 0.015, about 0.010, or about 0.005.

[0267] The water activity values are determined by the method of Saturated Aqueous Solutions (Multon, “Techniques d’Analyse E De Controle Dans Les Industries Agroalimentaires” APRIA (1981)) or by direct measurement using a viable Robotronic BT hygrometer or other hygrometer or hygroscope.

[0268] In some embodiments, the microbial composition comprises at least two different microbes, and wherein the at least two microbes are present in the composition at a ratio of 1:2, 1:3, 1:3, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:21, 1:22, 1:23, 1:24, 1:25, 1:26, 1:27, 1:28, 1:29, 1:30, 1:40, 1:50, 1:60, 1:100, 1:125, 1:150, 1:175, or 1:200 or the inverse thereof. In some embodiments, the microbial composition comprises at least three different microbes, and wherein the three microbes are present in the composition at a ratio of 1:2:1, 1:1:2, 2:2:1, 1:3:1, 1:1:3, 3:1:1, 3:3:1, 1:5:1, 1:1:5, 5:1:1, 5:5:1, or 1:5:5. Encapsulation Compositions

[0269] In some embodiments, the microbes or microbial compositions of the disclosure are encapsulated in an encapsulating composition. An encapsulating composition protects the 2026204768   19 Jun 2026 microbes from external stressors prior to entering the gastrointestinal tract of beef cattle. In some embodiments, external stressors include thermal and physical stressors associated with pelleting and extrusion. In some embodiments, external stressors include chemicals present in the compositions. Encapsulating compositions further create an environment that may be beneficial to the microbes, such as minimizing the oxidative stresses of an aerobic environment on anaerobic microbes. See Kalsta et al. (US 5,104,662A), Ford (US 5,733,568A), and Mosbach and Nilsson (US 4,647,536A) for encapsulation compositions of microbes, and methods of encapsulating microbes.

[0270] In one embodiment, the compositions of the present disclosure exhibit a thermal tolerance, which is used interchangeably with heat tolerance and heat resistance. In one embodiment, thermal tolerant compositions of the present disclosure are tolerant of the high temperatures associated with feed manufacturing, mixing of feed and compositions of the present disclosure, storage in high heat environments, etc. In one embodiment, thermal tolerant compositions of the present disclosure are resistant to heat-killing and denaturation of the cell wall components and the intracellular environment.

[0271] In one embodiments, the encapsulation is a reservoir-type encapsulation. In one embodiment, the encapsulation is a matrix-type encapsulation. In one embodiment, the encapsulation is a coated matrix-type encapsulation. Burgain et al. (2011. J. Food Eng. 104:467483) discloses numerous encapsulation embodiments and techniques, all of which are incorporated by reference.

[0272] In some embodiments, the compositions of the present disclosure are encapsulated m one or more of the following: gellan gum, xanthan gum, K-Carrageenan, cellulose acetate phthalate, chitosan, starch, milk fat, whey protein, Ca-alginate, raftilose, raftiline, pectin, saccharide, glucose, maltodextrin, gum arabic, guar, seed flour, alginate, dextrins, dextrans, celluloase, gelatin, gelatin, albumin, casein, gluten, acacia gum, tragacanth, wax, paraffin, stearic acid, monodiglycerides, and diglycerides. In some embodiments, the compositions of the present disclosure are encapsulated by one or more of a polymer, carbohydrate, sugar, plastic, glass, polysaccharide, lipid, wax, oil, fatty acid, or glyceride. In one embodiment, the microbial composition is encapsulated by glucose. In one embodiment, the microbial composition is encapsulated by a glucose-containmg composition. In one embodiment, formulations of the 2026204768   19 Jun 2026 microbial composition comprise a glucose encapsulant. In one embodiment, formulations of the microbial composition comprise a glucose-encapsulated composition.

[0273] In some embodiments, the encapsulation of the compositions of the present disclosure is carried out by an extrusion, emulsification, coating, agglomeration, lyophilization, vitrification, foam drying, preservation by vaporization, vacuum-drying, or spray-drying.

[0274] In some embodiments, the encapsulated compositions of the present disclosure are vitrified. In some embodiments, encapsulation involves a process of drying a composition of the present disclosure in the presence of a substance which forms a glassy, amorphous solid state, a process known as vitrification, and in doing so encapsulates the composition. In some embodiments, the vitrified composition is protected from degradative conditions that would typically destroy or degrade microbes. Many common substances have the property of vitrification; that is , they will form a glassy solid state under certain conditions. Among these substances are several sugars, including sucrose and maltose, and other more complex compounds, such as polyvinyl pyrolidone (PVT). As any solution dries down, the molecules in the solution can either crystahze, or they can vitrify. A solute which has an extensive asymmetry may be a superior vitrifier, because of the hindrances to nucleation of crystals during drying. A substance that inhibits the crystallization of another substance may result in the combined substances forming a superior vitrification, such as raffinose in the presence of sucrose. See U.S. Patent Nos. 5,290,765 and 9,469,835.

[0275] In some embodiments, a microbial composition is produced that is encapsulated in a vitrified substance. The vitrified composition may be created by selecting a mixture including cells; combining said mixture with sufficient quantity of one or more vitrifying solutes to protect said mixture during drying and to inhibit destructive reactions; and drying said combination by exposing said combination to a desiccant, or desiccating conditions, at a temperature above that which said combination will freeze and below that at which said vitrifying solutes achieve the vitrified state, at approximately normal atmospheric pressure, until said combination is substantially dry.

[0276] In one embodiment, the encapsulating composition comprises microcapsules having a multiplicity of liquid cores encapsulated in a solid shell material. For purposes of the disclosure, a "multiplicity" of cores is defined as two or more. 2026204768   19 Jun 2026

[0277] A first category of useful fusible shell materials is that of normally solid fats, including fats which are already of suitable hardness and animal or vegetable fats and oils which are hydrogenated until their melting points are sufficiently high to serve the purposes of the present disclosure. Depending on the desired process and storage temperatures and the specific material selected, a particular fat can be either a normally solid or normally liquid material. The terms "normally solid” and "normally liquid” as used herein refer to the state of a material at desired temperatures for storing the resulting microcapsules. Since fats and hydrogenated oils do not, strictly speaking, have melting points, the term "melting point" is used herein to describe the minimum temperature at which the fusible material becomes sufficiently softened or liquid to be successfully emulsified and spray cooled, thus roughly corresponding to the maximum temperature at which the shell material has sufficient integrity to prevent release of the choline cores. "Melting point" is similarly defined herein for other materials which do not have a sharp melting point.

[0278] Specific examples of fats and oils useful herein (some of which require hardening) are as follows: animal oils and fats, such as beef tallow, mutton tallow, lamb tallow, lard or pork fat, fish oil, and sperm oil; vegetable oils, such as canola oil, cottonseed oil, peanut oil, corn oil, olive oil, soybean oil, sunflower oil, safflower oil, coconut oil, palm oil, linseed oil, tung oil, and castor oil; fatty acid monoglycerides and diglycerides; free fatty acids, such as stearic acid, palmitic acid, and oleic acid; and mixtures thereof. The above listing of oils and fats is not meant to be exhaustive, but only exemplary.

[0279] Specific examples of fatty acids include linoleic acid, y-linoleic acid, dihomo-y-linolenic acid, arachidonic acid, docosatetraenoic acid, vaccenic acid, nervonic acid, mead acid, erucic acid, gondoic acid, elaidic acid, oleic acid, palitoleic acid, stearidonic acid, eicosapentaenoic acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, nonadecyclic acid, arachidic acid, heneicosylic acid, behenic acid, tricosylic acid, lignoceric acid, pentacosylic acid, cerotic acid, heptacosyhc acid, montamc acid, nonacosylic acid, melissic acid, henatriacontylic acid, lacceroic acid, psyllic acid, geddic acid, ceroplastic acid, hexatriacontylic acid, heptatriacontanoic acid, and octatriacontanoic acid.

[0280] Another category of fusible materials useful as encapsulating shell materials is that of waxes. Representative waxes contemplated for use herein are as follows: animal waxes, such as 2026204768   19 Jun 2026 beeswax, lanolin, shell wax, and Chinese insect wax; vegetable waxes, such as carnauba, candelilla, bayberry, and sugar cane; mineral waxes, such as paraffin, microcrystalline petroleum, ozocerite, ceresin, and montan; synthetic waxes, such as low molecular weight polyolefin (e.g., CARBOWAX), and polyol ether-esters (e.g., sorbitol); Fischer-Tropsch process synthetic waxes; and mixtures thereof. Water-soluble waxes, such as CARBOWAX and sorbitol, are not contemplated herein if the core is aqueous.

[0281] Still other fusible compounds useful herein are fusible natural resins, such as rosin, balsam, shellac, and mixtures thereof.

[0282] In some embodiments, the microbes or microbial composition is embedded in a wax, such as the waxes described in the present disclosure.

[0283] In some embodiments, the microbes or microbial composition is embedded in wax balls. In some embodiments, the microbes or microbial composition is already encapsulated prior to being embedded in wax balls. In some embodiments, the wax balls are 10 microbes, 20 microns, 30 microns, 40 microns, 50 microns, 60 microns, 70 microns, 80 microns, 90 microns, 100 microns, 150 microns, 200 microns, 250 microns, 300 microns, 350 microns, 400 microns, 450 microns, 500 microns, 550 microns, 600 microns, 650 microns, 700 microns, 750 microns, 800 microns, 850 microns, 900 microns, 950 microns, or 1,000 microns.

[0284] In some embodiments, the wax balls are about 10 microbes, about 20 microns, about 30 microns, about 40 microns, about 50 microns, about 60 microns, about 70 microns, about 80 microns, about 90 microns, about 100 microns, about 150 microns, about 200 microns, about 250 microns, about 300 microns, about 350 microns, about 400 microns, about 450 microns, about 500 microns, about 550 microns, about 600 microns, about 650 microns, about 700 microns, about 750 microns, about 800 microns, about 850 microns, about 900 microns, about 950 microns, or about 1,000 microns.

[0285] In some embodiments, the wax balls are between 10-20 microns, 10-30 microns, 10-40 microns, 10-50 microns, 10-60 microns, 10-70 microns, 10-80 microns, 10-90 microns, 10-100 microns, 10-250 microns, 10-500 microns, 10-750 microns, 10-1,000 microns, 20-30 microns, 20-40 microns, 20-50 microns, 20-60 microns, 20-70 microns, 20-80 microns, 20-90 microns, 20-100 microns, 20-250 microns, 20-500 microns, 20-750 microns, 20-1,000 microns, 30-40 microns, 30-50 microns, 30-60 microns, 30-70 microns, 30-80 microns, 30-90 microns, 30-100 microns, 30-250 microns, 30-500 microns, 30-750 microns, 30-1,000 microns, 40-50 microns, 2026204768   19 Jun 2026 40-60 microns, 40-70 microns, 40-80 microns, 40-90 microns, 40-100 microns, 40-250 microns, 40-500 microns, 40-750 microns, 40-1,000 microns, 50-60 microns, 50-70 microns, 50-80 microns, 50-90 microns, 50-100 microns, 50-250 microns, 50-500 microns, 50-750 microns, 501,000 microns, 60-70 microns, 60-80 microns, 60-90 microns, 60-100 microns, 60-250 microns, 60-500 microns, 60-750 microns, 60-1,000 microns, 70-80 microns 70-90 microns, 70-90 microns, 70-100 microns, 70-250 microns, 70-500 microns, 70-750 microns, 70-1,000 microns, 80-90 microns, 80-100 microns, 80-250 microns, 80-500 microns, 80-500 microns, 80-750 microns, 80-1,000 microns, 90-100 microns, 90-250 microns, 90-500 microns, 90-750 microns, 90-1,000 microns, 100-250 microns, 100-500 microns, 100-750 microns, 100-1,000 microns, 250-500 microns, 250-750 microns, 250-1,000 microns, 500-750 microns, 500-1,000 microns, or 750-1,000 microns.

[0286] In some embodiments, the wax balls are between about 10-20 microns, about 10-30 microns, about 10-40 microns, about 10-50 microns, about 10-60 microns, about 10-70 microns, about 10-80 microns, about 10-90 microns, about 10-100 microns, about 10-250 microns, about 10-500 microns, about 10-750 microns, about 10-1,000 microns, about 20-30 microns, about 2040 microns, about 20-50 microns, about 20-60 microns, about 20-70 microns, about 20-80 microns, about 20-90 microns, about 20-100 microns, about 20-250 microns, about 20-500 microns, about 20-750 microns, about 20-1,000 microns, about 30-40 microns, about 30-50 microns, about 30-60 microns, about 30-70 microns, about 30-80 microns, about 30-90 microns, about 30-100 microns, about 30-250 microns, about 30-500 microns, about 30-750 microns, about 30-1,000 microns, about 40-50 microns, about 40-60 microns, about 40-70 microns, about 40-80 microns, about 40-90 microns, about 40-100 microns, about 40-250 microns, about 40-500 microns, about 40-750 microns, about 40-1,000 microns, about 50-60 microns, about 50-70 microns, about 50-80 microns, about 50-90 microns, about 50-100 microns, about 50-250 microns, about 50-500 microns, about 50-750 microns, about 50-1,000 microns, about 60-70 microns, about 60-80 microns, about 60-90 microns, about 60-100 microns, about 60-250 microns, about 60-500 microns, about 60-750 microns, about 60-1,000 microns, about 70-80 microns about 70-90 microns, about 70-90 microns, about 70-100 microns, about 70-250 microns, about 70-500 microns, about 70-750 microns, about 70-1,000 microns, about 80-90 microns, about 80-100 microns, about 80-250 microns, about 80-500 microns, about 80-500 microns, about 80-750 microns, about 80-1,000 microns, about 90-100 microns, about 90-250 2026204768   19 Jun 2026 microns, about 90-500 microns, about 90-750 microns, about 90-1,000 microns, about 100-250 microns, about 100-500 microns, about 100-750 microns, about 100-1,000 microns, about 250500 microns, about 250-750 microns, about 250-1,000 microns, about 500-750 microns, about 500-1,000 microns, or about 750-1,000 microns.

[0287] Various adjunct materials are contemplated for incorporation in fusible materials according to the present disclosure. For example, antioxidants, light stabilizers, dyes and lakes, flavors, essential oils, anti-caking agents, fillers, pH stabilizers, sugars (monosaccharides, disaccharides, trisaccharides, and polysaccharides) and the like can be incorporated in the fusible material in amounts which do not diminish its utility for the present disclosure.

[0288] The core material contemplated herein constitutes from about 0.1% to about 50%, about 1% to about 35%. or about 5% to about 30% by weight of the microcapsules. In some embodiments, the core material contemplated herein constitutes no more than about 30% by weight of the microcapsules. In some embodiments, the core material contemplated herein constitutes about 5% by weight of the microcapsules. The core material is contemplated as either a liqui d or solid at contemplated storage temperatures of the microcapsules.

[0289] The cores may include other additives well-known in the pharmaceutical art, including edible sugars, such as sucrose, glucose, maltose, fructose, lactose, cellobiose, monosaccharides, disaccharides, trisaccharides, and polysaccharides, and mixtures thereof; artificial sweeteners, such as aspartame, saccharin, cyclamate salts, and mixtures thereof; edible acids, such as acetic acid (vinegar), citric acid, ascorbic acid, tartaric acid, and mixtures thereof; edible starches, such as corn starch; hydrolyzed vegetable protein; water-soluble vitamins, such as Vitamin C; watersoluble medicaments; water-soluble nutritional materials, such as ferrous sulfate; flavors; salts; monosodium glutamate; antimicrobial agents, such as sorbic acid; antimycotic agents, such as potassium sorbate, sorbic acid, sodium benzoate, and benzoic acid; food grade pigments and dyes; and mixtures thereof. Other potentially useful supplemental core materials will be apparent to those of ordinary skill in the art.

[0290] Emulsifying agents may be employed to assist in the formation of stable emulsions. Representative emulsifying agents include glyceryl monostearate, polysorbate esters, ethoxylated mono- and diglycerides, and mixtures thereof.

[0291] For ease of processing, and particularly to enable the successful formation of a reasonably stable emulsion, the viscosities of the core material and the shell material should be 2026204768   19 Jun 2026 similar at the temperature at which the emulsion is formed. In particular, the ratio of the viscosity of the shell to the viscosity of the core, expressed in centipoise or comparable units, and both measured at the temperature of the emulsion, should be from about 22:1 to about 1:1, desirably from about 8:1 to about 1:1, and preferably from about 3:1 to about 1:1. A ratio of 1:1 would be ideal, but a viscosity ratio within the recited ranges is useful.

[0292] Encapsulating compositions are not limited to microcapsule compositions as disclosed above. In some embodiments encapsulating compositions encapsulate the microbial compositions in an adhesive polymer that can be natural or synthetic without toxic effect. In some embodiments, the encapsulating composition may be a matrix selected from sugar matrix, gelatin matrix, polymer matrix, silica matrix, starch matrix, foam matrix, glass / glassy matrix etc. See Pirzio et al. (U.S. Patent 7,488,503). In some embodiments, the encapsulating composition may be selected from polyvinyl acetates; polyvinyl acetate copolymers; ethylene vinyl acetate (EVA) copolymers; polyvinyl alcohols; polyvinyl alcohol copolymers; celluloses, including ethylcelluloses, methylcelluloses, hydroxymethylcelluloses, hydroxypropylcelluloses and carboxymethylcellulose; polyvinylpyrolidones; polysaccharides, including starch, modified starch, dextrins, maltodextrins, alginate and chitosans; monosaccharides; fats; fatty acids, including oils; proteins, including gelatin and zeins; gum arabics; shellacs; vinylidene chloride and vinylidene chloride copolymers; calcium lignosulfonates; acrylic copolymers; polyvinyl acrylates; polyethylene oxide; acrylamide polymers and copolymers; poly hydroxy ethyl acrylate, methylacrylamide monomers; and polychloroprene.

[0293] In some embodiments, the encapsulating compositions comprise at least one layer of encapsulation. In some embodiments, the encapsulating compositions comprise at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 layers of encapsulation'encapsulants.

[0294] In some embodiments, the encapsulating compositions comprise at least two layers of encapsulation. In some embodiments, each layer of encapsulation confers a different characteristic to the composition. In some embodiments, no two consecutive layers confer the same characteristic. In some embodiments, at least one layer of the at least two layers of encapsulation confers thermostability, shelf stability, ultraviolet resistance, moisture resistance, 2026204768   19 Jun 2026 hydrophobicity, hydrophilicity, lipophobicity, lipophilicity, pH stability, acid resistance, and base resistance.

[0295] In some embodiments, the encapsulating compositions comprise two layers of encapsulation; the first layer confers thermostability and / or shelf stability, and the second layer provides pH resistance.

[0296] In some embodiments, the encapsulating layers confer a timed release of the microbial composition held in the center of the encapsulating layers. In some embodiments, the greater the number of layers confers a greater amount of time before the microbial composition is exposed, post administration.

[0297] In some embodiments, the encapsulating shell of the present disclosure can be up to 10pm, 20pm, 30pm, 40pm, 50pm, 60pm, 70pm, 80pm, 90pm, 100pm, 110pm, 120pm, 130pm, 140pm, 150pm, 160pm, 170pm, 180pm, 190pm, 200pm, 210pm, 220pm, 230pm, 240pm, 250pm, 260pm, 270pm, 280pm, 290pm, 300pm, 310pm, 320pm, 330pm, 340pm, 350pm, 360pm, 370pm, 380pm, 390pm, 400pm, 410pm, 420pm, 430pm, 440pm, 450pm, 460pm, 470pm, 480pm, 490pm, 500pm, 510pm, 520pm, 530pm, 540pm, 550pm, 560pm, 570pm, 580pm, 590pm, 600pm, 610pm, 620pm, 630pm, 640pm, 650pm, 660pm, 670pm, 680pm, 690pm, 700pm, 710pm, 720pm, 730pm, 740pm, 750pm, 760pm, 770pm, 780pm, 790pm, 800pm, 810pm, 820pm, 830pm, 840pm, 850pm, 860pm, 870pm, 880pm, 890pm, 900pm, 910pm, 920pm, 930pm, 940pm, 950pm, 960pm, 970pm, 980pm, 990pm, 1000pm, 1010pm, 1020pm, 1030pm, 1040pm, 1050pm, 1060pm, 1070pm, 1080pm, 1090pm, 1100pm, 1110pm, 1120pm, 1130pm, 1140pm, 1150pm, 1160pm, 1170 pm, 1180pm, 1190pm, 1200pm, 1210pm, 1220pm, 1230pm, 1240pm, 1250pm, 1260pm, 1270pm, 1280pm, 1290pm, 1300pm, 1310pm, 1320pm, 1330pm, 1340pm, 1350pm, 1360pm, 1370pm, 1380pm, 1390pm, 1400pm, 1410pm, 1420pm, 1430pm, 1440pm, 1450pm, 1460pm, 1470pm, 1480pm, 1490pm, 1500pm, 1510pm, 1520pm, 1530pm, 1540pm, 1550pm, 1560pm, 1570pm, 1580pm, 1590pm, 1600pm, 1610pm, 1620pm, 1630pm, 1640pm, 1650pm, 1660pm, 1670pm, 1680pm, 1690pm, 1700pm, 1710pm, 1720pm, 1730pm, 1740pm, 1750pm, 1760pm, 1770pm, 1780pm, 1790pm, 1800pm, 1810pm, 1820pm, 1830pm, 1840pm, 1850pm, 1860pm, 1870pm, 1880pm, 1890pm, 1900pm, 1910pm, 1920pm, 1930pm, 1940pm, 1950pm, 1960pm, 1970pm, 1980pm, 1990pm, 2000pm, 2010pm, 2020pm, 2030pm, 2040pm, 2050pm, 2060pm, 2070pm, 2080pm, 2090pm, 2100pm, 2110pm, 2120pm, 2130pm, 2140pm, 2150pm, 2160pm, 2170pm, 2180pm, 2190pm, 2200pm, 2210pm, 2026204768   19 Jun 2026 2220gm, 2230pm, 2240j.tm, 2250pm, 2260pm, 2270,11m, 2280pm, 2290pm, 2300pm, 2310pm, 2320pm, 2330,um, 2340pm, 2350pm, 2360pm, 2370pm, 2380pm, 2390pm, 2400pm, 2410pm, 2420pm, 2430pm, 2440pm, 2450pm, 2460pm, 2470,um, 2480,um, 2490,um, 2500pm, 2510pm, 2520,um, 2530,um, 2540pm, 2550pm, 2560pm, 2570pm, 2580pm, 2590pm, 2600pm, 261 0pm, 2620pm, 2630pm, 2640pm, 2650pm, 2660pm, 2670pm, 2680pm, 2690pm, 2700pm, 2710pm, 2720pm, 2730pm, 2740pm, 2750pm, 2760pm, 2770pm, 2780pm, 2790pm, 2800pm, 2810pm, 2820pm, 2830pm, 2840pm, 2850pm, 2860pm, 2870pm, 2880pm, 2890pm, 2900pm, 2910pm, 2920pm, 2930pm, 2940pm, 2950pm, 2960pm, 2970pm, 2980pm, 2990pm, or 3000pm thick.

[0298] In some embodiments, the encapsulation composition of the present disclosure possesses a water activity (aw) of less than 0.750, 0.700, 0.650, 0.600, 0.550, 0.500, 0.475, 0.450, 0.425, 0.400, 0.375, 0.350, 0.325, 0.300, 0.275, 0.250, 0.225, 0.200, 0.190, 0.180, 0.170, 0.160, 0.150, 0.140, 0.130, 0.120, 0.110, 0.100, 0.095, 0.090, 0.085, 0.080, 0.075, 0.070, 0.065, 0.060, 0.055, 0.050, 0.045, 0.040, 0.035, 0.030, 0.025, 0.020, 0.015, 0.010, or 0.005.

[0299] In some embodiments, the encapsulation composition of the present disclosure possesses a water activity (aw) of less than about 0.750, about 0.700, about 0.650, about 0.600, about 0.550, about 0.500, about 0.475, about 0.450, about 0.425, about 0.400, about 0.375, about 0.350, about 0.325, about 0.300, about 0.275, about 0.250, about 0.225, about 0.200, about 0.190, about 0.180, about 0.170, about 0.160, about 0.150, about 0.140, about 0.130, about 0.120, about 0.110, about 0.100, about 0.095, about 0.090, about 0.085, about 0.080, about 0.075, about 0.070, about 0.065, about 0.060, about 0.055, about 0.050, about 0.045, about 0.040, about 0.035, about 0.030, about 0.025, about 0.020, about 0.015, about 0.010, or about 0.005.

[0300] In one embodiment, the microbe(s) are first dried by spray dry, lyophilization, or foam drying along with excipients that may include one or more sugars, sugar alcohols, disaccharides, trisaccharides, polysaccharides, salts, ammo acids, amino acid salts, or polymers,

[0301] In some embodiments, the microbes or compositions comprising the microbes are milled to a size of 10 microns, 20 microns, 30 microns, 40 microns, 50 microns, 60 microns, 70 microns, 80 microns, 90 microns, 100 microns, 150 microns, 200 microns, 250 microns, 300 microns, 350 microns, 400 microns, 450 microns, 500 microns, 550 microns, 600 microns, 650 microns, 700 microns, 750 microns, 800 microns, 850 microns, 900 microns, 950 microns, or 1,000 microns. 2026204768   19 Jun 2026

[0302] In some embodiments, the microbes or compositions comprising the microbes are milled to a size of about 10 microbes, about 20 microns, about 30 microns, about 40 microns, about 50 microns, about 60 microns, about 70 microns, about 80 microns, about 90 microns, about 100 microns, about 150 microns, about 200 microns, about 250 microns, about 300 microns, about 350 microns, about 400 microns, about 450 microns, about 500 microns, about 550 microns, about 600 microns, about 650 microns, about 700 microns, about 750 microns, about 800 microns, about 850 microns, about 900 microns, about 950 microns, or about 1,000 microns.

[0303] In some embodiments, the microbes or compositions comprising the microbes are milled to a size of between 10-20 microns, 10-30 microns, 10-40 microns, 10-50 microns, 10-60 microns, 10-70 microns, 10-80 microns, 10-90 microns, 10-100 microns, 10-250 microns, 10500 microns, 10-750 microns, 10-1,000 microns, 20-30 microns, 20-40 microns, 20-50 microns, 20-60 microns, 20-70 microns, 20-80 microns, 20-90 microns, 20-100 microns, 20-250 microns, 20-500 microns, 20-750 microns, 20-1,000 microns, 30-40 microns, 30-50 microns, 30-60 microns, 30-70 microns, 30-80 microns, 30-90 microns, 30-100 microns, 30-250 microns, 30500 microns, 30-750 microns, 30-1,000 microns, 40-50 microns, 40-60 microns, 40-70 microns, 40-80 microns, 40-90 microns, 40-100 microns, 40-250 microns, 40-500 microns, 40-750 microns, 40-1,000 microns, 50-60 microns, 50-70 microns, 50-80 microns, 50-90 microns, 50100 microns, 50-250 microns, 50-500 microns, 50-750 microns, 50-1,000 microns, 60-70 microns, 60-80 microns, 60-90 microns, 60-100 microns, 60-250 microns, 60-500 microns, 60750 microns, 60-1,000 microns, 70-80 microns 70-90 microns, 70-90 microns, 70-100 microns, 70-250 microns, 70-500 microns, 70-750 microns, 70-1,000 microns, 80-90 microns, 80-100 microns, 80-250 microns, 80-500 microns, 80-500 microns, 80-750 microns, 80-1,000 microns, 90-100 microns, 90-250 microns, 90-500 microns, 90-750 microns, 90-1,000 microns, 100-250 microns, 100-500 microns, 100-750 microns, 100-1,000 microns, 250-500 microns, 250-750 microns, 250-1,000 microns, 500-750 microns, 500-1,000 microns, or 750-1,000 microns.

[0304] In some embodiments, the microbes or compositions comprising the microbes are milled to a size of between about 10-20 microns, about 10-30 microns, about 10-40 microns, about 1050 microns, about 10-60 microns, about 10-70 microns, about 10-80 microns, about 10-90 microns, about 10-100 microns, about 10-250 microns, about 10-500 microns, about 10-750 microns, about 10-1,000 microns, about 20-30 microns, about 20-40 microns, about 20-50 microns, about 20-60 microns, about 20-70 microns, about 20-80 microns, about 20-90 microns, 2026204768   19 Jun 2026 about 20-100 microns, about 20-250 microns, about 20-500 microns, about 20-750 microns, about 20-1,000 microns, about 30-40 microns, about 30-50 microns, about 30-60 microns, about 30-70 microns, about 30-80 microns, about 30-90 microns, about 30-100 microns, about 30-250 microns, about 30-500 microns, about 30-750 microns, about 30-1,000 microns, about 40-50 microns, about 40-60 microns, about 40-70 microns, about 40-80 microns, about 40-90 microns, about 40-100 microns, about 40-250 microns, about 40-500 microns, about 40-750 microns, about 40-1,000 microns, about 50-60 microns, about 50-70 microns, about 50-80 microns, about 50-90 microns, about 50-100 microns, about 50-250 microns, about 50-500 microns, about 50750 microns, about 50-1,000 microns, about 60-70 microns, about 60-80 microns, about 60-90 microns, about 60-100 microns, about 60-250 microns, about 60-500 microns, about 60-750 microns, about 60-1,000 microns, about 70-80 microns about 70-90 microns, about 70-90 microns, about 70-100 microns, about 70-250 microns, about 70-500 microns, about 70-750 microns, about 70-1,000 microns, about 80-90 microns, about 80-100 microns, about 80-250 microns, about 80-500 microns, about 80-500 microns, about 80-750 microns, about 80-1,000 microns, about 90-100 microns, about 90-250 microns, about 90-500 microns, about 90-750 microns, about 90-1,000 microns, about 100-250 microns, about 100-500 microns, about 100750 microns, about 100-1,000 microns, about 250-500 microns, about 250-750 microns, about 250-1,000 microns, about 500-750 microns, about 500-1,000 microns, or about 750-1,000 microns.

[0305] In some embodiments, the microbes or compositions comprising the microbes are combined with a wax, fat, oil, fatty acid, or fatty alcohol, and spray congealed into beads of about 10 microbes, about 20 microns, about 30 microns, about 40 microns, about 50 microns, about 60 microns, about 70 microns, about 80 microns, about 90 microns, about 100 microns, about 150 microns, about 200 microns, about 250 microns, about 300 microns, about 350 microns, about 400 microns, about 450 microns, about 500 microns, about 550 microns, about 600 microns, about 650 microns, about 700 microns, about 750 microns, about 800 microns, about 850 microns, about 900 microns, about 950 microns, or about 1,000 microns.

[0306] In some embodiments, the microbes or compositions comprising the microbes are combined with a wax, fat, oil, fatty acid, or fatty alcohol, and spray congealed into beads of between 10-20 microns, 10-30 microns, 10-40 microns, 10-50 microns, 10-60 microns, 10-70 microns, 10-80 microns, 10-90 microns, 10-100 microns, 10-250 microns, 10-500 microns, 10- 2026204768   19 Jun 2026 750 microns, 10-1,000 microns, 20-30 microns, 20-40 microns, 20-50 microns, 20-60 microns, 20-70 microns, 20-80 microns, 20-90 microns, 20-100 microns, 20-250 microns, 20-500 microns, 20-750 microns, 20-1,000 microns, 30-40 microns, 30-50 microns, 30-60 microns, 3070 microns, 30-80 microns, 30-90 microns, 30-100 microns, 30-250 microns, 30-500 microns, 30-750 microns, 30-1,000 microns, 40-50 microns, 40-60 microns, 40-70 microns, 40-80 microns, 40-90 microns, 40-100 microns, 40-250 microns, 40-500 microns, 40-750 microns, 401,000 microns, 50-60 microns, 50-70 microns, 50-80 microns, 50-90 microns, 50-100 microns, 50-250 microns, 50-500 microns, 50-750 microns, 50-1,000 microns, 60-70 microns, 60-80 microns, 60-90 microns, 60-100 microns, 60-250 microns, 60-500 microns, 60-750 microns, 601,000 microns, 70-80 microns 70-90 microns, 70-90 microns, 70-100 microns, 70-250 microns, 70-500 microns, 70-750 microns, 70-1,000 microns, 80-90 microns, 80-100 microns, 80-250 microns, 80-500 microns, 80-500 microns, 80-750 microns, 80-1,000 microns, 90-100 microns, 90-250 microns, 90-500 microns, 90-750 microns, 90-1,000 microns, 100-250 microns, 100-500 microns, 100-750 microns, 100-1,000 microns, 250-500 microns, 250-750 microns, 250-1,000 microns, 500-750 microns, 500-1,000 microns, or 750-1,000 microns.

[0307] In some embodiments, the microbes or compositions comprising the microbes are combined with a wax, fat, oil, fatty acid, or fatty alcohol, and spray congealed into beads of between about 10-20 microns, about 10-30 microns, about 10-40 microns, about 10-50 microns, about 10-60 microns, about 10-70 microns, about 10-80 microns, about 10-90 microns, about 10100 microns, about 10-250 microns, about 10-500 microns, about 10-750 microns, about 101,000 microns, about 20-30 microns, about 20-40 microns, about 20-50 microns, about 20-60 microns, about 20-70 microns, about 20-80 microns, about 20-90 microns, about 20-100 microns, about 20-250 microns, about 20-500 microns, about 20-750 microns, about 20-1,000 microns, about 30-40 microns, about 30-50 microns, about 30-60 microns, about 30-70 microns, about 30-80 microns, about 30-90 microns, about 30-100 microns, about 30-250 microns, about 30-500 microns, about 30-750 microns, about 30-1,000 microns, about 40-50 microns, about 4060 microns, about 40-70 microns, about 40-80 microns, about 40-90 microns, about 40-100 microns, about 40-250 microns, about 40-500 microns, about 40-750 microns, about 40-1,000 microns, about 50-60 microns, about 50-70 microns, about 50-80 microns, about 50-90 microns, about 50-100 microns, about 50-250 microns, about 50-500 microns, about 50-750 microns, about 50-1,000 microns, about 60-70 microns, about 60-80 microns, about 60-90 microns, about 2026204768   19 Jun 2026 60-100 microns, about 60-250 microns, about 60-500 microns, about 60-750 microns, about 601,000 microns, about 70-80 microns about 70-90 microns, about 70-90 microns, about 70-100 microns, about 70-250 microns, about 70-500 microns, about 70-750 microns, about 70-1,000 microns, about 80-90 microns, about 80-100 microns, about 80-250 microns, about 80-500 microns, about 80-500 microns, about 80-750 microns, about 80-1,000 microns, about 90-100 microns, about 90-250 microns, about 90-500 microns, about 90-750 microns, about 90-1,000 microns, about 100-250 microns, about 100-500 microns, about 100-750 microns, about 1001,000 microns, about 250-500 microns, about 250-750 microns, about 250-1,000 microns, about 500-750 microns, about 500-1,000 microns, or about 750-1,000 microns.

[0308] In some embodiments, the microbes or compositions comprising the microbes are combined with a wax, fat, oil, fatty acid, or fatty alcohol as well as a water-soluble polymer, salt, polysaccharide, sugar, polypeptide, protein, or sugar alcohol and spray congealed into beads, the size of which are described herein. In some embodiments, the water-soluble polymer, salt, polysaccharide, sugar, or sugar alcohol serves as a disintegrant. In some embodiments, the dismtegrant forms pores once the beads are dispersed in the rumen of the animal.

[0309] In some embodiments, the composition of the water-soluble polymer, salt, polysaccharide, sugar, polypeptide, protein, or sugar alcohol is modified such that the disintegrant dissolves within 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 minutes of being administered. In some embodiments, the composition of the water-soluble polymer, salt, polysaccharide, sugar, polypeptide, protein, or sugar alcohol is modified such that the disintegrant dissolves within about 1, about 5, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, or about 60 minutes of being administered.

[0310] In some embodiments, the composition of the water-soluble polymer, salt, polysaccharide, sugar, polypeptide, protein, or sugar alcohol is modified such that the disintegrant dissolves within 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, or 12 hours of being administered. In some embodiments, the composition of the water-soluble polymer, salt, polysaccharide, sugar, polypeptide, protein, or sugar alcohol is modified such that the disintegrant dissolves within about 1, about 1.5, about 2, about 2.5, about 3, about 3.5, about 4, about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, about 7.5, about 2026204768   19 Jun 2026 8, about 8.5, about 9, about 9.5, about 10, about 10.5, about 11, about 11.5, or about 12 hours of being administered.

[0311] In some embodiments, the composition of the water-soluble polymer, salt, polysaccharide, sugar, polypeptide, protein, or sugar alcohol is modified such that the disintegrant dissolves at a temperature of at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 °C. In some embodiments, the composition of the water-soluble polymer, salt, polysaccharide, sugar, polypeptide, protein, or sugar alcohol is modified such that the disintegrant dissolves at a temperature of at least about 10, least about 11, least about 12, least about 13, least about 14, least about 15, least about 16, least about 17, least about 18, least about 19, least about 20, least about 21, least about 22, least about 23, least about 24, least about 25, least about 26, least about 27, least about 28, least about 29, least about 30, least about 31, least about 32, least about 33, least about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, least about 45, least about 46, least about 47, least about 48, least about 49, or least about 50 °C.

[0312] In some embodiments, the composition of the water-soluble polymer, salt, polysaccharide, sugar, polypeptide, protein, or sugar alcohol is modified such that the disintegrant dissolves at a pH of at least 3.8, 3.9, 4. 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9 or 10.0. In some embodiments, the composition of the water-soluble polymer, salt, polysaccharide, sugar, polypeptide, protein, or sugar alcohol is modified such that the disintegrant dissolves at a pH of at least about 3.8, least about 3.9, least about 4. least about 4.1, least about 4.2, least about 4.3, least about 4.4, least about 4.5, least about 4.6, least about 4.7, least about 4.8, least about 4.9, least about 5.0, least about 5.1, least about 5.2, least about 5.3, least about 5.4, least about 5.5, least about 5.6, least about 5.7, least about 5.8, least about 5.9, least about 6.0, least about 6.2, least about 6.3, least about 6.4, least about 6.5, least about 6.6, least about 6.7, least about 6.8, least about 6.9, least about 7.0, least about 7.1, least about 7.2, least about 7.3, least about 7.4, least about 7.5, least about 7.6, least about 7.7, least about 7.8, least about 7.9, least about 8.0, least about 8.1, least about 8.2, least about 8.3, least about 8.4, least about 8.5, least about 8.6, least about 8.7, least about 8.8, least about 8.9, least about 9.0, 2026204768   19 Jun 2026 least about 9.1, least about 9.2, least about 9.3, least about 9.4, least about 9.5, least about 9.6, least about 9.7, least about 9.8, least about 9.9, or least about 10.0.

[0313] In some embodiments, the microbes or compositions comprising the microbes are coated with a polymer, a polysaccharide, sugar, sugar alcohol, gel, wax, fat, fatty alcohol, or fatty acid

[0314]

[0315] In some embodiments, the microbes or compositions comprising the microbes are coated with a polymer, a polysaccharide, sugar, sugar alcohol, gel, wax, fat, fatty alcohol, or fatty acid.

[0316] In some embodiments, the coating of the microbes or compositions comprising the microbes is modified such that the coating dissolves within 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 minutes of being administered. In some embodiments, the coating of the microbes or compositions comprising the microbes is modified such that the coating dissolves within about 1, about 5, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, or about 60 minutes of being administered.

[0317] In some embodiments, the coating of the microbes or compositions comprising the microbes is modified such that the coating dissolves within 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, or 12 hours of being administered. In some embodiments, the coating of the microbes or compositions comprising the microbes is modified such that the coating dissolves within about 1, about 1.5, about 2, about 2.5, about 3, about 3.5, about 4, about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, about 7.5, about 8, about 8.5, about 9, about 9.5, about 10, about 10.5, about 11, about 11.5, or about 12 hours of being administered.

[0318] In some embodiments, the coating of the microbes or compositions comprising the microbes is modified such that the coating dissolves at a temperature of at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 °C. In some embodiments, the coating of the microbes or compositions comprising the microbes is modified such that the coating dissolves at a temperature of at least about 10, least about 11, least about 12, least about 13, least about 14, least about 15, least about 16, least about 17, least about 18, least about 19, least about 20, least about 21, least about 22, least about 23, least about 24, least about 25, least about 26, least about 2026204768   19 Jun 2026 27, least about 28, least about 29, least about 30, least about 31, least about 32, least about 33, least about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, least about 45, least about 46, least about 47, least about 48, least about 49, or least about 50 °C.

[0319] In some embodiments, the coating of the microbes or compositions comprising the microbes is modified such that the coating dissolves at a pH of at least 3.8, 3.9, 4. 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9 or 10.0. In some embodiments, the coating of the microbes or compositions comprising the microbes is modified such that the coating dissolves at a pH of at least about 3.8, least about 3.9, least about 4. least about 4.1, least about 4.2, least about 4.3, least about 4.4, least about 4.5, least about 4.6, least about 4.7, least about 4.8, least about 4.9, least about 5.0, least about 5.1, least about 5.2, least about 5.3, least about 5.4, least about 5.5, least about 5.6, least about 5.7, least about 5.8, least about 5.9, least about 6.0, least about 6.2, least about 6.3, least about 6.4, least about 6.5, least about 6.6, least about 6.7, least about 6.8, least about 6.9, least about 7.0, least about 7.1, least about 7.2, least about 7.3, least about 7.4, least about 7.5, least about 7.6, least about 7.7, least about 7.8, least about 7.9, least about 8.0, least about 8.1, least about 8.2, least about 8.3, least about 8.4, least about 8.5, least about 8.6, least about 8.7, least about 8.8, least about 8.9, least about 9.0, least about 9.1, least about 9.2, least about 9.3, least about 9.4, least about 9.5, least about 9.6, least about 9.7, least about 9.8, least about 9.9, or least about 10.0. Animal Feed

[0320] In some embodiments, compositions of the present disclosure are mixed with animal feed. In some embodiments, animal feed may be present in various forms such as pellets, capsules, granulated, powdered, mash, liquid, semi-liquid, or mixed rations(s).

[0321] In some embodiments, compositions of the present disclosure are mixed into the premix at the feed mill (e.g., Carghill or Western Millin), alone as a standalone premix, and / or alongside other feed additives such as MONENSIN, vitamins, etc. In one embodiment, compositions of the 2026204768   19 Jun 2026 present disclosure are mixed into the feed itself. In one embodiment, the compositions of the present disclosure are mixed into the feed at the feed mill.

[0322] In some embodiments, feed of the present disclosure may be supplemented with water, premix or premixes, forage, beans (e.g., whole, cracked, or ground), grains (e.g., whole, cracked, or ground), bean- or grain-based oils, bean- or grain-based meals, bean- or gram-based haylage or silage, bean- or grain-based syrups, fatty acids, sugar alcohols (e.g., polyhydric alcohols), commercially available formula feeds, oyster shells and those of other bivalves, and mixtures thereof.

[0323] In some embodiments, forage encompasses hay, haylage, and silage. In some embodiments, hays include grass hays (e.g., sudangrass, orchardgrass, or the like), alfalfa hay, and clover hay. In some embodiments, haylages include grass haylages, sorghum haylage, and alfalfa haylage. In some embodiments, silages include maize, oat, wheat, alfalfa, clover, and the like.

[0324] In some embodiments, premix or premixes may be utilized in the feed. Premixes may comprise micro-ingredients such as vitamins, minerals, amino acids; chemical preservatives; pharmaceutical compositions such as antibiotics, ionophores, and other medicaments; fermentation products, and other ingredients. In some embodiments, premixes are blended into the feed.

[0325] In some embodiments, the feed may include feed concentrates such as soybean hulls, soybean oils, sugar beet pulp, molasses, high protein soybean meal, ground corn, shelled corn, cornflakes, wheat midds, distiller gram, cottonseed hulls, rumen-bypass protein, rumen-bypass fat,and grease. See Luhman (U.S. Publication US20150216817A1), Anderson et al. (U.S. Patent 3,484,243), Porter and Luhman (U.S. Patent 9,179,694B2), Iritani et al. (U.S. Patent 6,090,416), Axelrod et al. (U.S. Publication US20060127530A1), and Katsumi et al. (U.S. Patent 5,741,508) for animal feed and animal feed supplements capable of use in the present compositions and methods.

[0326] In some embodiments, feed occurs as a compound, which includes, in a mixed composition capable of meeting the basic dietary needs, the feed itself, vitamins, minerals, ammo acids, and other necessary components. Compound feed may further comprise premixes. 2026204768   19 Jun 2026

[0327] In some embodiments, microbial compositions of the present disclosure may be mixed with ammai feed, premix, and / or compound feed. Individual components of the animal feed may be mixed with the microbial compositions prior to feeding to beef cattle. The microbial compositions of the present disclosure may be applied into or on a premix, into or on a feed, and / or into or on a compound feed.

[0328] In some embodiments, microbial compositions of the present disclosure may be mixed with animal feed, premix, and / or compound feed at various stages of animal adaptation to the step-up or finishing diet.

[0329] In some embodiments, microbial compositions of the present disclosure are mixed with feed and microingredients. Microingredients include liquid fat blends, glycerin, rumensm, monensin, vitamins, tylan, optaflex, melengesterol acetate, minerals, and amino acids. In some embodiments, the mixing of feed, microbial compositions of the present disclosure, and microingredients is performed at the feedlot.

[0330] In some embodiments, cattle begin a step up or a starting ration. As used herein, a “step-up diet” or “starting ration” is a diet fed to feedlot cattle as a transition to the high grain content of the finishing diet. In some embodiments, the step-up diet may involve one or more step-up diets that ease the cattle into the transition to the finishing diet. In some embodiments, the step-up diet is formulated to slowly increase the amount of high energy feed in the diet while mitigating gastrointestinal distress and the effects of rapid onset acidosis. In some embodiments, the cattle are fed a single type of step-up diet. In some embodiments, the cattle are fed multiple varieties of step-up diets, increasing the amount of high energy feed with each iteration of the step up diet variety'. In some embodiments, the cattle are fed at least one step up diet, wherein the subsequent diets are different from each of those step up diets that follow. In some embodiments, the cattle are fed at least two different step up diets. In some embodiments, the cattle are fed at least three different step up diets.

[0331] As used herein, a “finishing diet” is a concentrated high-energy diet (often high-grain) fed to cattle on a feedlot to rapidly bring the cattle up to get them to market weight by the time the cattle are rendered. In some embodiments, the finishing diet may result in liver disease, liver abscesses, and / or acidosis. 2026204768   19 Jun 2026

[0332] In some embodiments, the microbial compositions of the present disclosure are mixed with step-up diets. In some embodiments, the microbial compositions of the present disclosure are mixed with finishing diets. Administration of Microbial Compositions

[0333] In some embodiments, the microbial compositions of the present disclosure are administered to cattle via the oral route. In some embodiments the microbial compositions are administered via a direct injection route into the gastrointestinal tract. In further embodiments, the direct injection administration delivers the microbial compositions directly to the rumen. In some embodiments, the microbial compositions of the present disclosure are administered to animals anally. In further embodiments, anal administration is in the form of an inserted suppository.

[0334] In some embodiments, the microbial composition is administered in a dose volume comprising a total of, or at least, 1ml, 2ml, 3ml, 4ml, 5ml, 6ml, 7ml, 8ml, 9ml, 10ml, 11 ml, 12ml, 13ml, 14ml, 15ml, 16ml, 17ml, 18ml, 19ml, 20ml, 21ml, 22ml, 23ml, 24ml, 25ml, 26ml, 27ml, 28ml, 29ml, 30ml, 31ml, 32ml, 33ml, 34ml, 35ml, 36ml, 37ml, 38ml, 39ml, 40ml, 41m, 42ml, 43ml, 44ml, 45ml, 46ml, 47ml, 48ml, 49ml, 50ml, 60ml, 70ml, 80ml, 90ml, 100ml, 200ml, 300ml, 400ml, 500ml, 600ml, 700ml, 800ml, 900ml, or 1,000ml.

[0335] In some embodiments, the microbial composition is administered in a dose comprising a total of, or at least, 10ls, 1017, 1016, 1015, 1014, 1013, 1012, 10n, IO:". IO9, 10s, 107, 106, 105, IO4, 10J, or 102 microbial cells.

[0336] In some embodiments, the microbial compositions are mixed with feed, and the administration occurs through the ingestion of the microbial compositi ons along with the feed. In some embodiments, the dose of the microbial composition is administered such that there exists 102 to 1012, 103 to 10i2, 104 to 1012, 105 to 1012, 106 to 1012, 107 to 1012, 10s to 1012, IO9 to 1012, 1010to 1012, 10n to 1012, 102to 10H, 103 to 1011, 104 to 10n, 105 to 10n, 106 to 10n, 107 to 1011, 10s to 1011, 105 to 1011, 1010to 1011, 102 to 1010, 103 to 1010, 104 to 1010, 105 to 1010, 106 to 1010, 107 to 1010, 10s to 1010, 109 to 1010, 102 to 109, 103 to 109, 104 to 109, 105 to 109, 106 to 109, 107 to 109, 10s to 109, 102 to 10s, 103 to 10s, 104 to 10s, 105 to 10s, 106 to 10s, 107 to 10s, 102 to 107, 103 to 107, 104 to 107, 105 to 107, 106 to 107, 102 to 106, 103 to 106, 104 to 106, 105 to 106,102to 105, 103 2026204768   19 Jun 2026 to 105, 104 to 105, 102 to 104, 103 to 104, 102 to 103, 1012, 1011, 1010, 109, 108, 107, 106, 105, 104, 103, or 102 total microbial cells per gram or milliliter of the composition.

[0337] In some embodiments, the administered dose of the microbial composition comprises 102 to 1018, 103 to 1018, 104 to 10!8, 105 to 1018, 106to 1018, 107 to 1018, 108to 1018, 109 to 1018, 1010to 1018, 1011 to 1018, 10!2to IO18, 1013 to IO18, 10i4to 10!8, 1015 to 10!8, 1016to 1018, 10!7to 10i8, 102 to 1012, 103to IO12, 104to IO12, 105to 10!2, 106 to 1012, 107to 10i2, 108to 10!2, 109to 10!2, 10f°to 1012, 10ff to IO12, 102 to 10!!, 103 to 1011, 104 to 10JJ, 105 to 10l!, 106 to 10n, 107 to 10!!, 108 to 10n, 109 to 10!!, 10f°to 10", 102 to 1010, 103 to 10w, 104 to 10!°, 105 to 10!°, 106 to 10i0, 107 to 10l°, 108 to 10i0, 109 to 1010, 102 to 109, 103 to 109, 104 to 109, 105 to 109, 106 to 109, 107 to 109, 108 to 109, 102 to 108, 103 to 108, 104 to 108, 105 to 108, 106 to 10s, 107 to 108, 102 to 107, 103 to 107, 104 to 107, 105 to 107, 106 to 107, 102 to 106, 103 to 106, 104 to 106, 105 to 106,102to 105, 103 to 105, 104 to 105, 102to 104, 103 to 104, 102 to 103, 1018, 10i7, 10!6, 1015, 1014, 1013, 1012, 1011, 1010, 109, 108, 10 ', 106, 105, 104, 103, or 102 total microbial cells.

[0338] In some embodiments, the composition is administered 1 or more times per day. In some aspects, the composition is administered with food each time the animal is fed. In some embodiments, the composition is administered 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 3 to 4, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 4 to 5, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, 6 to 10, 6 to 9, 6 to 8, 6 to 7, 7 to 10, 7 to 9, 7 to 8,8 to 10, 8 to 9, 9 to 10, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times per day.

[0339] In some embodiments, the microbial composition is administered 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 3 to 4, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 4 to 5, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, 6 to 10, 6 to 9, 6 to 8, 6 to 7, 7 to 10, 7 to 9, 7 to 8,8 to 10, 8 to 9, 9 to 10, 1,2, 3, 4, 5, 6, 7, 8, 9, or 10 times per week.

[0340] In some embodiments, the microbial composition is administered 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 3 to 4, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 4 to 5, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, 6 to 10, 6 to 9, 6 to 8, 6 to 7, 7 to 10, 7 to 9, 7 to 8,8 to 10, 8 to 9, 9 to 10, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times per month. 2026204768   19 Jun 2026

[0341] In some embodiments, the microbial composition is administered 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 3 to 4, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 4 to 5, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, 6 to 10, 6 to 9, 6 to 8, 6 to 7, 7 to 10, 7 to 9, 7 to 8,8 to 10, 8 to 9, 9 to 10, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times per year.

[0342] In some embodiments, the microbial composition is administered to animals throughout the entire time they are on the feedlot. In some embodiments, the microbial composition is administered to animals only during a portion of time while they are on the feedlot. In some embodiments, the microbial composition is administered only during the grower phase. In some embodiments, the microbial composition is administered only during the time when animals are in the receiving pen. In some embodiments, the microbial composition is administered only when the animals are receiving vaccinations and / or treatments. In some embodiments, the microbial composition is administered only when the animals are on a step up diet or when being adapted to a high grain diet. In some embodiments, the microbial composition is administered only when the animals are on a finisher diet or a high grain diet.

[0343] In some embodiments, the microbial composition is administered during the grower phase, when animals are in the receiving pen, when animals are receiving vaccinations and / or treatments, when animals are being adapted to a high grain diet or are on a step up diet, and / or when the animals are on a finisher diet or a high grain diet.

[0344] In some embodiments, an animal entering the feed lot receives at least one microbial composition prior to entering the feed lot. In some embodiments, an animal on the feed lot receives a microbial composition that is different from the first at least one microbial composition. In further embodiments, an animal on the feed lot receives a microbial composition that is different from the first and second at least one microbial composition.

[0345] In some embodiments, the type of diet fed to the animal corresponds with the type of microbial composition administered to the animal. In some embodiments, a grazing or grass / hay-fed animal will receive a first microbial composition. In further embodiments, the same animal fed a different diet will receive a second microbial composition, wherein the first microbial composition is different from the second microbial composition. In some embodiments, the same animal fed yet a different diet will receive a third microbial composition, wherein the first microbial composition is different from the second and third microbial compositions. In some 2026204768   19 Jun 2026 embodiments, the same animal fed yet a different diet will receive a fourth microbial composition, wherein the first microbial composition is different from the second, third, and fourth microbial compositions. In some embodiments, the same animal fed yet a different diet will receive a fifth microbial composition, wherein the first microbial composition is different from the second, third, fourth, and fifth microbial compositions.

[0346] In some embodiments, the feed can be uniformly coated with one or more layers of the microbes and / or microbial compositions disclosed herein, using conventional methods of mixing, spraying, or a combination thereof through the use of treatment application equipment that is specifically designed and manufactured to accurately, safely, and efficiently apply coatings. Such equipment uses various types of coating technology such as rotary coaters, drum coaters, fluidized bed techniques, spouted beds, rotary mists, or a combination thereof. Liquid treatments such as those of the present disclosure can be applied via either a spinning “atomizer” disk or a spray nozzle, which evenly distributes the microbial composition onto the feed as it moves though the spray pattern. In some aspects, the feed is then mixed or tumbled for an additional period of time to achieve additional treatment distribution and drying.

[0347] In some embodiments, the feed coats of the present disclosure can be up to 10pm, 20pm, 30pm, 40pm, 50pm, 60pm, 70pm, 80pm, 90pm, 100pm, 110pm, 120pm, 130pm, 140pm, 150pm, 160pm, 170pm, 180pm, 190pm, 200pm, 210pm, 220pm, 230pm, 240pm, 250pm, 260pm, 270pm, 280pm, 290pm, 300pm, 310pm, 320 pm, 330pm, 340pm, 350pm, 360 pm, 370pm, 380pm, 390 pm, 400pm, 410pm, 420pm, 430 pm, 440pm, 450pm, 460pm, 470pm, 480pm, 490pm, 500 pm, 510pm, 520pm, 530pm, 540pm, 550pm, 560pm, 570pm, 580 pm, 590 pm, 600pm, 610pm, 620pm, 630pm, 640pm, 650pm, 660pm, 670pm, 680pm, 690pm, 700pm, 710pm, 720pm, 730pm, 740pm, 750 pm, 760pm, 770pm, 780pm, 790pm, 800pm, 810 pm, 820pm, 830pm, 840pm, 850pm, 860pm, 870pm, 880pm, 890pm, 900pm, 910pm, 920pm, 930pm, 940pm, 950pm, 960pm, 970pm, 980pm, 990pm, 1000pm, 1010pm, 1020pm, 1030pm, 1040pm, 1050pm, 1060pm, 1070pm, 1080pm, 1090pm, 1100pm, 1110pm, 1120pm, 1130pm, 1140pm, 1150pm, 1160pm, 1170pm, 1180pm, 1190pm, 1200pm, 1210pm, 1220pm, 1230pm, 1240pm, 1250pm, 1260pm, 1270pm, 1280pm, 1290pm, 1300pm, 1310pm, 1320pm, 1330pm, 1340pm, 1350pm, 1360pm, 1370pm, 1380pm, 1390pm, 1400pm, 1410pm, 1420pm, 1430pm, 1440pm, 1450pm, 1460pm, 1470pm, 1480pm, 1490pm, 1500pm, 1510pm, 1520pm, 1530pm, 1540pm, 1550pm, 1560pm, 1570pm, 1580pm, 1590pm, 1600pm, 1610pm, 1620pm, 2026204768   19 Jun 2026 1630|im, 1640|im, 1650jrm, 1660pm, 1670pm, 1680,11m, 1690pm, 1700pm, 1710pm, 1720pm, 1730pm, 1740,um, 1750pm, 1760pm, 1770pm, 1780pm, 1790pm, 1800pm, 1810pm, 1820pm, 1830pm, 1840pm, 1850pm, 1860pm, 1870pm, 1880pm, 1890,um, 1900,um, 1910pm, 1920pm, 1930,um, 1940,um, 1950pm, 1960pm, 1970pm, 1980pm, 1990pm, 2000pm, 2010pm, 2020pm, 2030pm, 2040pm, 2050pm, 2060pm, 2070pm, 2080pm, 2090pm, 2100pm, 2110pm, 2120pm, 2130pm, 2140pm, 2150pm, 2160pm, 2170pm, 2180pm, 2190pm, 2200pm, 2210pm, 2220pm, 2230pm, 2240pm, 2250pm, 2260pm, 2270pm, 2280pm, 2290pm, 2300pm, 2310pm, 2320pm, 2330pm, 2340pm, 2350pm, 2360pm, 2370pm, 2380pm, 2390pm, 2400pm, 2410pm, 2420pm, 2430pm, 2440pm, 2450pm, 2460pm, 2470pm, 2480pm, 2490pm, 2500pm, 2510pm, 2520pm, 2530pm, 2540pm, 2550pm, 2560pm, 2570pm, 2580pm, 2590pm, 2600pm, 2610pm, 2620pm, 2630pm, 2640pm, 2650pm, 2660pm, 2670pm, 2680pm, 2690pm, 2700pm, 2710pm, 2720pm, 2730pm, 2740pm, 2750pm, 2760pm, 2770pm, 2780pm, 2790pm, 2800pm, 2810pm, 2820pm, 2830pm, 2840pm, 2850pm, 2860pm, 2870pm, 2880pm, 2890pm, 2900pm, 2910pm, 2920pm, 2930pm, 2940pm, 2950pm, 2960pm, 2970pm, 2980pm, 2990pm, or 3000pm thick. [034S] In some embodiments, the microbial cells can be coated freely onto any number of compositions or they can be formulated in a liquid or solid composition before being coated onto a composition. For example, a solid composition comprising the microorganisms can be prepared by mixing a solid carrier with a suspension of the spores until the solid carriers are impregnated with the spore or cell suspension. This mixture can then be dried to obtain the desired particles.

[0349] In some other embodiments, it is contemplated that the solid or liquid microbial compositions of the present disclosure further contain functional agents e.g., activated carbon, minerals, vitamins, and other agents capable of improving the quality of the products or a combination thereof.

[0350] Methods of coating and compositions in use of said methods that are known in the art can be particularly useful when they are modified by the addition of one of the embodiments of the present disclosure. Such coating methods and apparatus for their application are disclosed in, for example: U.S. Pat. Nos. 8,097,245 and 7,998,502; and PCT Pat. App. Pub, Nos. WO 2008 / 076975, WO 2010 / 138522, WO 2011 / 094469, WO 2010 / 111347, and WO 2010 / 111565 each of which is incorporated by reference herein. 2026204768   19 Jun 2026

[0351] In some embodiments, the microbes or microbial compositions of the present disclosure exhibit a synergistic effect, on one or more of the traits described herein, in the presence of one or more of the microbes or microbial compositions coming into contact with one another. The synergistic effect obtained by the taught methods can be quantified, for example, according to Colby’s formula (i.e., (E) = X+Y - (X*Y / 100)). See Colby, RS., “Calculating Synergistic and Antagonistic Responses of Herbicide Combinations,” 1967. Weeds. Vol. 15, pp. 20-22, incorporated herein by reference in its entirety. Thus, “synergistic” is intended to reflect an outcome / parameter / effect that has been increased by more than an additive amount.

[0352] In some embodiments, the microbes or microbial compositions of the present disclosure may be administered via bolus. In one embodiment, a bolus (e.g., capsule containing the composition) is inserted into a bolus gun, and the bolus gun is inserted into the buccal cavity and / or esophagas of the animal, followed by the release / injection of the bolus into the animal’s digestive tract. In one embodiment, the bolus gun / applicator is a BOVIKALC bolus gun / applicator. In another embodiment, the bolus gun / applicator is a QUADRICAL gun / applicator.

[0353] In some embodiments, the microbes or microbial compositions of the present disclosure may be administered via drench. In one embodiment, the drench is an oral drench. A drench administration comprises utilizing a drench kit / applicator / syringe that injects / releases a liquid comprising the microbes or microbial compositions into the buccal cavity and / or esophagus of the animal.

[0354] In some embodiments, the microbes or microbial compositions of the present disclosure may be administered in a time-released fashion. The composition may be coated in a chemical composition, or may be contained in a mechanical device or capsule that releases the microbes or microbial compositions over a period of time instead all at once. In one embodiment, the microbes or microbial compositions are administered to an animal in a time-release capsule. In one embodiment, the composition may be coated in a chemical composition, or may be contained in a mechanical device or capsule that releases the microbes or microbial compositions all at once a period of time hours post ingestion.

[0355] In some embodiments, one microbe composition is administered one or more times when the animals are on a step up diet, and a different microbe composition is administered one or more times when the animals are on a finishing diet. In some embodiments, one microbe 2026204768   19 Jun 2026 composition is administered one or more times when the animals are on a step up diet, a different microbe composition is administered one or more times when the animals are on the first thirty days of the finishing diet, and yet a different microbe composition is administered one or more times when the animals have been on the finishing diet for greater than thirty days.

[0356] In some embodiments, one microbe composition is administered one or more times while the animals exhibit signs of acidosis, and different microbe composition is administered one or more times once the signs of acidosis have abated. In some embodiments, a microbe composition is administered to animals that do not exhibit signs of acidosis, and a different microbe composition is administered if the animals exhibit signs of acidosis.

[0357] In some embodiments, the microbes or microbial compositions are administered in a time-released fashion between 1 to 5, 1 to 10, 1 to 15, 1 to 20, 1 to 24, 1 to 25, 1 to 30, 1 to 35, 1 to 40, 1 to 45, 1 to 50, 1 to 55, 1 to 60, 1 to 65, 1 to 70, 1 to 75, 1 to 80, 1 to 85, 1 to 90, 1 to 95, or 1 to 100 hours.

[0358] In some embodiments, the microbes or microbial compositions are administered in a time-released fashion between 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 1 to 11, 1 to 12, 1 to 13, 1 to 14, 1 to 15, 1 to 16, 1 to 17, 1 to 18, 1 to 19, 1 to 20, 1 to 21, 1 to 22, 1 to 23, 1 to 24, 1 to 25, 1 to 26, 1 to 27, 1 to 28, 1 to 29, or 1 to 30 days. Microorganisms

[0359] As used herein the term “microorganism” should be taken broadly. It includes, but is not limited to, the two prokaryotic domains, Bacteria and Archaea, as well as eukaryotic fungi, protozoa, and viruses.

[0360] By way of example, the microorganisms may include species of the genera of: Fibrobacter, Saccharofermentans, Bacillus, Spirochaeta, Bacteroides, Lachnospiracea incertae sedis, Clostridium XLVa, Ruminococcus, Butyricimonas, Olsenella, Acidaminococcus, Parabacteroides, Clostridum sensu stricto, Oribacterium, Pseudoflavonifractor, Treponema, Rhodobacter, Fluviicola, Succiniclasticum, Solobaclerium, Veillonella, Cellulosimicrobium, Cupriavidus, Megasphaera, Suednivibrio, Oscillibacter, Pseudomonas, Corynebacterium, Adlercreutzia, Dorea, Roseburia, Anaerovibrio, Sporosarcina, Streptomyces, Syntrophococcus, Butyrivibrio, Lachnobacterium, Pyramidobacler, Coprococcus, Ruminobacter, Thermobifidia, 2026204768   19 Jun 2026 Papillibacter, Aquimarina, Propioniciclava, Staphylococcus, Mogibacterium, Pseudobutyrivibrio, Asteroleplasma, Turicibacter, Aggregalibacter, Brevundimonas, Phascolarctobacterium, and Sphingobium..

[0361] In certain embodiments, the microorganism is unculturable. This should be taken to mean that the microorganism is not known to be culturable or is difficult to culture using methods known to one skilled in the art.

[0362] In one embodiment, the microbes are obtained from animals (e.g., mammals, reptiles, birds, and the like), soil (e.g., rhizosphere), air, water (e.g., marine, freshwater, wastewater sludge), sediment, oil, plants (e.g., roots, leaves, stems), agricultural products, and extreme environments (e.g., acid mine drainage or hydrothermal systems). In a further embodiment, microbes obtained from marine or freshwater environments such as an ocean, river, or lake. In a further embodiment, the microbes can be from the surface of the body of water, or any depth of the body of water (e.g., a deep sea sample).

[0363] The microorganisms of the disclosure may be isolated in substantially pure or mixed cultures. They may be concentrated, diluted, or provided in the natural concentrations in which they are found in the source material. For example, microorganisms from saline sediments may be isolated for use in this disclosure by suspending the sediment in fresh water and allowing the sediment to fall to the bottom. The water containing the bulk of the microorganisms may be removed by decantation after a suitable period of settling and either administered to the GI tract of beef cattle, or concentrated by filtering or centrifugation, diluted to an appropriate concentration and administered to the GI tract of beef cattle with the bulk of the salt removed. By way of further example, microorganisms from mineralized or toxic sources may be similarly treated to recover the microbes for application to beef cattle to minimize the potential for damage to the animal,

[0364] In another embodiment, the microorganisms are used in a crude form, in which they are not isolated from the source material in which they naturally reside. For example, the microorganisms are provided in combination with the source material in which they reside; for example, fecal matter, rumen content, rumen fluid, or other composition found in the gastrointestinal tract. In this embodiment, the source material may include one or more species of microorganisms. 2026204768   19 Jun 2026

[0365] In some embodiments, a mixed population of microorganisms is used in the methods of the disclosure.

[0366] In embodiments of the disclosure where the microorganisms are isolated from a source material (for example, the material in which they naturally reside), any one or a combination of a number of standard techniques which will be readily known to skilled persons may be used. However, by way of example, these in general employ processes by which a solid or liquid culture of a single microorganism can be obtained in a substantially pure form, usually by physical separation on the surface of a solid microbial growth medium or by volumetric dilutive isolation into a liquid microbial growth medium. These processes may include isolation from dry material, liquid suspension, slurries or homogenates in which the material is spread in a thin layer over an appropriate solid gel growth medium, or serial dilutions of the material made into a sterile medium and inoculated into liquid or solid culture media.

[0367] While not essential, in one embodiment, the material containing the microorganisms may be pre-treated prior to the isolation process in order to either multiply all microorganisms in the material, remove certain microorganisms in the material, and / or shift the distribution of microorganisms in the material. Microorganisms can then be isolated from the enriched materials as disclosed above.

[0368] In certain embodiments, as mentioned herein before, the microorganism(s) may be used in crude form and need not be isolated from an animal or a media. For example, feces, or growth media which includes the microorganisms identified to be of benefit to increased feed efficiencymay be obtained and used as a crude source of microorganisms for the next round of the method or as a crude source of microorganisms at the conclusion of the method. For example, fresh feces could be obtained and optionally processed. Microbiome Shift and Abundance of Microbes

[0369] In some embodiments, the microbiome of beef cattle, including the rumen microbiome comprises a diverse arrive of microbes with a wide variety of metabolic capabilities. The microbiome is influenced by a range of factors including diet, variations in animal metabolism, and breed, among others. Most cattle diets are plant-based and rich in complex polysaccharides 2026204768   19 Jun 2026 that enrich the gastrointestinal microbial community for microbes capable of breaking down specific polymeric components in the diet such as cellulose, hemicellulose, lignin, etc. The end products of primary degradation sustain a chain of microbes that ultimately produce a range of organic acids together with hydrogen and carbon dioxide. Because of the complex and interlinked nature of the microbiome, changing the diet and thus substrates for primary degradation may have a cascading effect on gut microbial metabolism, with changes in both the organic acid profiles and the methane levels produced, thus impacting the quality and quantity of animal production and or the products produced by the animal. See Menezes et al. (2011. FEMS Microbiol. Ecol. 78(2):256-265.)

[0370] In some aspects, the present disclosure is drawn to administering microbial compositions described herein to modulate or shift the microbiome of beef cattle.

[0371] In some embodiments, the microbiome is shifted through the administration of one or more microbes to one or more sections of the gastrointestinal tract. In some embodiments, the microbiome is shifted through the administration of one or more microbes to the rumen. In further embodiments, the one or more microbes are those selected from Table 1 and / or Table 2. In some embodiments, the microbiome shift or modulation includes a decrease or loss of specific microbes that were present prior to the administration of one or more microbes of the present disclosure. In some embodiments, the microbiome shift or modulation includes an increase in microbes that were present prior to the administration of one or more microbes of the present disclosure. In some embodiments, the microbiome shift or modulation includes a gain of one or more microbes that were not present prior to the administration of one or more microbes of the present disclosure. In a further embodiment, the gain of one or more microbes is a microbe that was not specifically included in the administered microbial composition.

[0372] In some embodiments, the administration of microbes of the present disclosure results in a sustained modulation of the microbiome such that the administered microbes are present in the microbiome for a period of at least 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 3 to 4, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 4 to 5, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, 6 to 10, 6 to 9, 6 to 8, 6 to 7, 7 to 10, 7 to 9, 7 to 8,8 to 10, 8 to 9, 9 to 10, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days. 2026204768   19 Jun 2026

[0373] In some embodiments, the administration of microbes of the present disclosure results in a sustained modulation of the microbiome such that the administered microbes are present in the microbiome for a period of at least 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 3 to 4, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 4 to 5, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, 6 to 10, 6 to 9, 6 to 8, 6 to 7, 7 to 10, 7 to 9, 7 to 8,8 to 10, 8 to 9, 9 to 10, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks.

[0374] In some embodiments, the administration of microbes of the present disclosure results in a sustained modulation of the microbiome such that the administered microbes are present in the microbiome for a period of at least 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, 3 to 4, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 4 to 5, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, 6 to 10, 6 to 9, 6 to 8, 6 to 7, 7 to 10, 7 to 9, 7 to 8,8 to 10, 8 to 9, 9 to 10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

[0375] In some embodiments, the presence of the administered microbes are detected by sampling the gastrointestinal tract and using primers to amplify the 16S or 18S rDNA sequences, or the ITS rDNA sequences of the administered microbes. In some embodiments, the administered microbes are one or more of those selected from Table 1 and / or Table 2. In some embodiments, the administered microbes are one or more of those comprising rDNA sequences selected from SEQ ID NO: 1-5993.

[0376] In some embodiments, the microbiome of beef cattle is measured by amplifying polynucleotides collected from gastrointestinal samples, wherein the polynucleotides may be 16S or 18S rDNA fragments, or ITS rDNA fragments of microbial rDNA. In one embodiment, the microbiome is fingerprinted by a method of denaturing gradient gel electrophoresis (DGGE) wherein the amplified rDNA fragments are sorted by where they denature, and form a unique banding pattern in a gel that may be used for comparing the microbiome of the same beef cattle over time or the microbiomes of multiple. In another embodiment, the microbiome is fingerprinted by a method of terminal restriction fragment length polymorphism (T-RFLP), wherein labelled PCR fragments are digested using a restriction enzyme and then sorted by size. In a further embodiment, the data collected from the T-RFLP method is evaluated by nonmetric 2026204768   19 Jun 2026 multidimensional scaling (nMDS) ordination and PERMANOVA statistics identify differences in microbiomes, thus allowing for the identification and measurement of shifts in the microbiome. See also Shanks et al. (2011. Appl. Environ. Microbiol. 77(9):2992-3001), Petri el al. (2013. PLOS one. 8(12):e83424), and Menezes et al. (2011. FEMS Microbiol. Ecol. 78(2):256-265.)

[0377] In some embodiments, administration of one or more microbial compositions results in a shift in the microbiome that increases the number and / or type of carbon dioxide fixing microbes. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that increases the number and / or type of carbon dioxide fixing microbes by at least 0.5%, at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, or at least 700%. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that increases the number and / or type of carbon dioxide fixing microbes by at least about 0.5%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 300%, at least about 400%, at least about 500%, at least about 600%, or at least about 700%.

[0378] In some embodiments, administration of one or more microbial compositions results in a shift in the microbiome that decreases the number and / or type of methanogenic microbes. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that reduces the number and / or type of methanogenic microbes by at least 0.5%, at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that decreases the number and / or type of methanogenic microbes by at least about 0.5%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 2026204768   19 Jun 2026 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95%.

[0379] In some embodiments, administration of one or more microbial compositions results in a shift in the microbiome that decreases the number and / or type of lactate producing microbes. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that reduces the number and / or type of lactate producing microbes by at least 0.5%, at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that decreases the number and / or type of lactate producing microbes by at least about 0.5%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95%. 103801 In some embodiments, administration of one or more microbial compositions results in a shift in the microbiome that increases the number and / or type of lactate degrading microbes. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that increases the number and / or type of lactate degrading microbes by at least 0.5%, at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45? / ), at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95? / ), at least 100%, at least 200%, at least 300%, at least 400? / , at least 500%, at least 600%, or at least 700%. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that increases the number and / or type of lactate degrading microbes by at least about 0.5%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, 2026204768   19 Jun 2026 at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 300%, at least about 400%, at least about 500%, at least about 600%, or at least about 700%.

[0381] In some embodiments, administration of one or more microbial compositions results in a shift in the microbiome that increases the number and / or type of volatile fatty acid (VFA) -producing microbes. In some embodiments, the VFAs include acetate, butyrate, propionate, isobutyrate, isovalerate, and valerate. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that increases the number and / or type of VFA-producing microbes by at least 0.5%, at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, or at least 700%. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that increases the number and / or type of VFA-producing microbes by at least about 0.5%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 300%, at least about 400%, at least about 500%, at least about 600%, or at least about 700%.

[0382] In some embodiments, administration of one or more microbial compositions results in a shift in the microbiome that increases the number and / or type of microbes that are utilized as protein sources for the animal. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that increases the number and / or type of microbes that are utilized as protein sources for the animal by at least 0.5%, at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, or at least 700%. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that 2026204768   19 Jun 2026 increases the number and / or type of microbes that are utilized as protein sources for the animal by at least about 0.5%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 300%, at least about 400%, at least about 500%, at least about 600%, or at least about 700%.

[0383] In some embodiments, administration of one or more microbial compositions results in a shift in the microbiome that increases the number and / or type of vitamin synthesizing microbes. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that increases the number and / or type of vitamin synthesizing microbes by at least 0.5%, at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, or at least 700%. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that increases the number and / or type of vitamin synthesizing microbes by at least about 0.5%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65? / ), at least about 70 / 1), at least about 75 / 1), at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100? / ), at least about 200%, at least about 300%, at least about 400%, at least about 500%, at least about 600%, or at least about 700%.

[0384] In some embodiments, administration of one or more microbial compositions results in a shift in the microbiome that reduces the overall alpha diversity of the microbial community. In some embodiments, administration of one or more microbial composition results in a shift in the microbiome that reduces the overall alpha diversity of the microbial community by at least 0.5%, at least l?zo, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, or at least 700%. In some 2026204768   19 Jun 2026 embodiments, administration of one or more microbial composition results in a shift in the microbiome that reduces the overall alpha diversity of the microbial community by at least about 0.5%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, at least about 200%, at least about 300%, at least about 400%, at least about 500%, at least about 600%, or at least about 700%.

[0385] In some embodiments, the administration of microbes of the present disclosure results in a modulation or shift of the microbiome which further results in a desired phenotype or improved trait. Cattle Microbial Compositional Diversity

[0386] Bovine in a commercial settings have been found to exhibit a high degree of animal-to-animal variability in terms of the microbial diversify of the rumen. The increased variability of the microbial compositions of the rumen may lead to a lower ability to reach a stable microbial composition. Lower variability in turn results in a considerable difference in health, weight, and other attributes that affect commercial viability of the animal. See Shabat SKB et al. (ISME J 10:2958 2972.)

[0387] In some embodime...

Claims

1. A ruminant supplement capable of treating or preventing acidosis or bloat in a ruminant, comprising:a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993: andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to treat or prevent acidosis or bloat in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

2. A ruminant supplement capable of treating or preventing acidosis or bloat in a ruminant, comprising:a) a purified population of bacteria selected from:(i) SuccinMbrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO :86, and / or(in) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in anamount effective to treat or prevent acidosis or bloat in a ruminant administered the supplement,as compared to a ruminant not administered the supplement.

3. The ruminant supplement of claim 1, wherein at least one of the bacteria are selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97%2026204768   19 Jun 2026identical to SEQ ID NO:86, and / or(iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

4. The ruminant supplement of claim 2, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:75,(li) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO :86, and / or(iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 13.

5. The ruminant supplement of claim 2, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence comprising SEQ ID NO:75,(li) Prevotella bacteria with a 16S nucleic acid sequence comprising SEQ IDNO :86, and / or(iii) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 13.

6. The ruminant supplement of claim 2, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria deposited as B-67550.(li) Prevotella bacteria deposited as B-67552, and / or(iii) Bacteroides bacteria deposited as B-67555.

7. The ruminant supplement of claim 2, further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.2026204768   19 Jun 20268. The ruminant supplement of claim 2, further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

9. The ruminant supplement of claim 2, further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

10. The ruminant supplement of claim 2, wherein the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, plastic, glass, polysaccharide, lipid, wax, oil, fatty acid, or glyceride.

11. The ruminant supplement of claim 10, wherein the encapsulated bacteria are vitrified.

12. The ruminant supplement of claim 10, wherein the encapsulated bacteria are furtherencapsulated in a wax, fat, fatty acid, fatty alcohol, or glyceride.

13. The ruminant supplement of claim 2, wherein the purified population of bacteria are in the form of spores.

14. The ruminant supplement of claim 13, wherein the spores are spray dried.

15. The ruminant supplement of claim 13, formulated as a tablet, capsule, pill, feed additive,food ingredient, food additive, food preparation, food supplement, consumable solution, consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof.

16. A method for treating or preventing acidosis or bloat in a ruminant, comprising: administering to a ruminant an effective amount of a ruminant supplement comprising:2026204768   19 Jun 2026a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to treat or prevent acidosis or bloat in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

17. A method for treating or preventing acidosis or bloat in a ruminant, comprising: administering to a ruminant an effective amount of a ruminant supplement comprising:a) a purified population of bacteria selected from:(i) Succimvibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to treat or prevent acidosis or bloat in a ruminant administered the supplement, as compared to a ruminant not administered the supplement,18. The method of claim 16, wherein at least one of the bacteria are selected from:(i) Succinhibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

19. The method of claim 17, wherein the purified population of bacteria is selected from:2026204768   19 Jun 2026(1) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO :75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:86, and / or(lii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 13.

20. The method of claim 17, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence comprising SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence comprising SEQ ID NO:86, and / or(in) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 13.

21. The method of claim 17, wherein the purified population of bacteria is selected from :(i) Succinivibrio bacteria deposited as B-67550.(ii) Prevotella bacteria deposited as B-67552, and / or (in) Bacteroides bacteria deposited as B-67555.

22. The method of claim 17, wherein the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

23. The method of claim 17, wherein the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.2026204768   19 Jun 202624. The method of claim 17, wherein the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

25. The method of claim 17, wherein the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, sugar alcohol, surfactant, plastic, glass, polysaccharide, lipid, wax, oil, fatty acid, ammo acid, or glyceride.

26. The method of claim 25, wherein the encapsulated bacteria are vitrified.

27. The method of claim 25, wherein the encapsulated bacteria are further encapsulated in awax, fat, fatty' acid, fatty alcohol, or glyceride.

28. The method of claim 17, wherein the purified population of bacteria are in the form of spores.

29. The method of claim 28, wherein the spores are spray dried.

30. The method of claim 28, formulated as a tablet, capsule, pill, feed additive, foodingredient, food additive, food preparation, food supplement, consumable solution, consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof,31. The method of claim 17, wherein the ruminant supplement is administered to a ruminant orally.

32. The method of claim 31, wherein the ruminant is a cow or a steer.

33. The method of claim 31, wherein the ruminant is fed a step-up diet.

34. The method of claim 31, wherein the ruminant is fed a finishing diet.2026204768   19 Jun 202635. A ruminant supplement capable of decreasing the amount of carbon dioxide and / or carbonic acid in the rumen of a ruminant, comprising:a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ IDNO: 1-5993: andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to decrease the amount of carbon dioxide and / or carbonic acid in the rumen of a ruminant administered the supplement,as compared to a ruminant not administered the supplement.

36. A ruminant supplement capable of decreasing the amount of carbon dioxide and / or carbonic acid in the rumen of a ruminant, comprising:a) a purified population of bacteria selected from:(i) Succimvibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(li) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(lii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to decrease the amount of carbon dioxide and / or carbonic acid in therumen of a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

37. The ruminant supplement of claim 35, wherein at least one of the bacteria are selected from:(i) Succinhibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,2026204768   19 Jun 2026(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

38. The ruminant supplement of claim 36, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO :86, and / or(in) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 13.

39. The ruminant supplement of claim 36, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence comprising SEQ IDNO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence comprising SEQ IDNO :86, and / or(lii) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO:13.

40. The ruminant supplement of claim 36, wherein the purified population of bacteria is selected from :(i) Succinivibrio bacteria deposited as B-67550.(ii) Prevotella bacteria deposited as B-67552, and / or(iii) Bacteroides bacteria deposited as B-67555.2026204768   19 Jun 202641. The ruminant supplement of claim 36, further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

42. The ruminant supplement of claim 36, further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

43. The ruminant supplement of claim 36, further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

44. The ruminant supplement of claim 36, wherein the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, sugar alcohol, surfactant, plastic, glass, polysaccharide, lipid, wax, oil, fatty acid, amino acid, or glyceride.

45. The ruminant supplement of claim 44, wherein the encapsulated bacteria are vitrified.

46. The ruminant supplement of claim 44, wherein the encapsulated bacteria are furtherencapsulated in a wax, fat, fatty acid, fatty alcohol, or glyceride.

47. The ruminant supplement of claim 36, wherein the purified population of bacteria are in the form of spores.

48. The ruminant supplement of claim 47, wherein the spores are spray dried.

49. The ruminant supplement of claim 47, formulated as a tablet, capsule, pill, feed additive,food ingredient, food additive, food preparation, food supplement, consumable solution, consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof.2026204768   19 Jun 202650. A method decreasing the amount of carbon dioxide and / or carbonic acid in the rumen of a ruminant, comprising:administering to a ruminant an effective amount of a ruminant supplement comprising:a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to decrease the amount of carbon dioxide and / or carbonic acid in the rumen of a ruminant administered the supplement,as compared to a ruminant not administered the supplement.

51. A method decreasing the amount of carbon dioxide and / or carbonic acid in the rumen of a ruminant, comprising:administering to a ruminant an effective amount of a ruminant supplement comprising:a) a purified population of bacteria selected from:(i) SuccinMbrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(in) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to decrease the amount of carbon dioxide and / or carbonic acid in the rumen of a ruminant administered the supplement,as compared to a ruminant not administered the supplement.

52. The method of claim 50, wherein at least one of the bacteria are selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,2026204768   19 Jun 2026(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

53. The method of claim 51, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO :86, and / or(hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 13.

54. The method of claim 51, wherein the purified population of bacteria is selected from :(i) Succinivibrio bacteria with a 16S nucleic acid sequence comprising SEQ ID NO :75,(ii) Prevotella. bacteria with a 16S nucleic acid sequence comprising SEQ IDNO:86, and / or(in) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 13.

55. The method of claim 51, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria deposited as B-67550.(ii) Prevotella bacteria deposited as B-67552, and / or(iii) Bacteroides bacteria deposited as B-67555.

56. The method of claim 51, wherein the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.2026204768   19 Jun 202657. The method of claim 51, wherein the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

58. The method of claim 51, wherein the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

59. The method of claim 51, wherein the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, sugar alcohol, surfactant, plastic, glass, polysaccharide, lipid, wax, oil, fatty' acid, ammo acid, or glyceride.

60. The method of claim 59, wherein the encapsulated bacteria are vitrified.

61. The method of claim 59, wherein the encapsulated bacteria are further encapsulated in awax, fat, fatty acid, fatty alcohol, or glyceride.

62. The method of claim 51, wherein the purified population of bacteria are in the form of spores.

63. The method of claim 62, wherein the spores are spray dried.

64. The method of claim 62, formulated as a tablet, capsule, pill, feed additive, foodingredient, food additive, food preparation, food supplement, consumable solution, consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof.

65. The method of claim 51, wherein the ruminant supplement is administered to a ruminant orally.

66. The method of claim 65 wherein the ruminant is a cow or a steer.2026204768   19 Jun 202667.    The method of claim 65, wherein the ruminant is fed a step-up diet.

68. The method of claim 65, wherein the ruminant is fed a finishing diet.

69. A ruminant supplement capable of increasing the amount of meat marbling in a ruminant, comprising:a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993: andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase the amount of meat marbling in the ruminant administered the supplement,as compared to a ruminant not administered the supplement.

70. A ruminant supplement capable of increasing the amount of meat marbling in a ruminant, comprising:a) a purified population of bacteria selected from:(i) Succinivihrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(in) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in anamount effective to increase the amount of meat marbling in the ruminant administered the supplement,as compared to a ruminant not administered the supplement.

71. The ruminant supplement of claim 69, wherein at least one of the bacteria are selected from:2026204768   19 Jun 2026(1) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO :75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(lii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

72. The ruminant supplement of claim 70, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO :75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO:86, and / or(lii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 13.

73. The ruminant supplement of claim 70, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence comprising SEQ ID NO :75,(ii) Prevotella bacteria with a 16S nucleic acid sequence comprising SEQ IDNO:86, and / or(in) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO:13.

74. The ruminant supplement of claim 70, wherein the purified population of bacteria is selected from :(i) Succinivibrio bacteria deposited as B-67550.(ii) Prevotella bacteria deposited as B-67552, and / or(lii) Bacteroides bacteria deposited as B-67555.2026204768   19 Jun 202675. The ruminant supplement of claim 70, further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

76. The ruminant supplement of claim 70, further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

77. The ruminant supplement of claim 70, further comprising a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

78. The ruminant supplement of claim 70, wherein the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, sugar alcohol, surfactant, plastic, glass, polysaccharide, lipid, wax, oil, fatty acid, amino acid, or glyceride.

79. The ruminant supplement of claim 78, wherein the encapsulated bacteria are vitrified.

80. The ruminant supplement of claim 78, wherein the encapsulated bacteria are furtherencapsulated in a wax, fat, fatty acid, fatty alcohol, or glyceride.

81. The ruminant supplement of claim 70, wherein the purified population of bacteria are in the form of spores.

82. The ruminant supplement of claim 81, wherein the spores are spray dried.

83. The ruminant supplement of claim 81, formulated as a tablet, capsule, pill, feed additive,food ingredient, food additive, food preparation, food supplement, consumable solution, consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof.2026204768   19 Jun 202684. A method increasing the amount of meat marbling in a ruminant, comprising: administering to a ruminant an effective amount of a ruminant supplement comprising:a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase the amount of meat marbling in the ruminant administered the supplement,as compared to a ruminant not administered the supplement.

85. A method increasing the amount of meat marbling in a ruminant, comprising: administering to a ruminant an effective amount of a ruminant supplement comprising:a) a purified population of bacteria selected from:(i) SuccinMbrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO :86, and / or(in) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase the amount of meat marbling in the ruminant administered the supplement,as compared to a ruminant not administered the supplement.

86. The method of claim 84, wherein at least one of the bacteria are selected from:(i) Succinhibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO :75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO :86, and / or2026204768   19 Jun 2026(lii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

87. The method of claim 85, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO :75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO :86, and / or(iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 99% identical to SEQ ID NO: 13.

88. The method of claim 85, wherein the purified population of bacteria is selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence comprising SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence comprising SEQ IDNO :86, and / or(iii) Bacteroides bacteria with a 16S nucleic acid sequence comprising SEQ ID NO: 13.

89. The method of claim 85, wherein the purified population of bacteria is selected from :(i) Succinivibrio bacteria deposited as B-67550.(ii) Prevotella bacteria deposited as B-67552, and / or(iii) Bacteroides bacteria deposited as B-67555.

90. The method of claim 85, wherein the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 97% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.2026204768   19 Jun 202691. The method of claim 85, wherein the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence that is at least about 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

92. The method of claim 85, wherein the ruminant supplement further comprises a purified population of bacteria that comprises bacteria with a 16S nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-5993.

93. The method of claim 85, wherein the purified population of bacteria are encapsulated in one or more of a polymer, carbohydrate, sugar, sugar alcohol, surfactant, plastic, glass, polysaccharide, lipid, wax, oil, fatty' acid, ammo acid, or glyceride.

94. The method of claim 93, wherein the encapsulated bacteria are vitrified.

95. The method of claim 93, wherein the encapsulated bacteria are further encapsulated in awax, fat, fatty acid, fatty alcohol, or glyceride.

96. The method of claim 85 wherein the purified population of bacteria are in the form of spores.

97. The method of claim 96, wherein the spores are spray dried,98.    The method of claim 96, formulated as a tablet, capsule, pill, feed additive, foodingredient, food additive, food preparation, food supplement, consumable solution, consumable spray additive, consumable solid, consumable gel, injection, bolus, or combinations thereof.

99. The method of claim 96, wherein the ruminant supplement is administered to a ruminant orally.

100. The method of claim 99 wherein the ruminant is a cow or a steer.2026204768   19 Jun 2026101.   The method of claim 99, wherein the ruminant is fed a step-up diet.

102. The method of claim 99, wherein the ruminant is fed a finishing diet.

103. The method of claim 85, wherein the increase in meat marbling is an increase of at least10%.

104. A ruminant supplement capable of increasing feed efficiency in a ruminant, comprising: a) a purified population of bacteria selected from any one or more bacteria comprising a16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993: andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase feed efficiency in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

105. A ruminant supplement capable of increasing feed efficiency in a ruminant, comprising: a) a purified population of bacteria selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(li) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(lii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase feed efficiency in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.2026204768   19 Jun 2026106. The ruminant supplement of claim 104, wherein at least one of the bacteria are selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO :75,(li) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO :86, and / or(hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

107. A method for increasing feed efficiency in a ruminant, comprising:administering to a ruminant an effective amount of a ruminant supplement comprising:a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase feed efficiency in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

108. A method increasing feed efficiency in a ruminant, comprising:administering to a ruminant an effective amount of a ruminant supplement comprising:a) a purified population of bacteria selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an2026204768   19 Jun 2026amount effective to increase feed efficiency in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

109. The method of claim 107, wherein at least one of the bacteria are selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(li) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO :86, and / or(ni) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

110. A ruminant supplement capable of increasing performance in a ruminant, comprising: a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993: andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase performance in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

111. A ruminant supplement capable of increasing performance in a ruminant, comprising: a) a purified population of bacteria selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an amount effective to increase performance in a ruminant administered the supplement,2026204768   19 Jun 2026as compared to a ruminant not administered the supplement.

112. The ruminant supplement of claim 104, wherein at least one of the bacteria are selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(li) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO :86, and / or(lii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

113. A ruminant supplement capable of reducing methane production and emission in a ruminant, comprising:a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993: andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in anamount effective to reduce methane production and emission in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

114. A ruminant supplement capable of reducing methane production and emission in a ruminant, comprising:a) a purified population of bacteria selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO :75,(li) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(lii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in an2026204768   19 Jun 2026amount effective to reduce methane production and emission in a ruminant administered the supplement,as compared to a ruminant not administered the supplement.

115. The ruminant supplement of claim 104, wherein at least one of the bacteria are selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(li) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO :86, and / or(hi) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

116. A ruminant supplement capable of reducing methane production and emission in a ruminant, comprising:a) a purified population of bacteria selected from any one or more bacteria comprising a 16S nucleic acid sequence that is at least about 97% identical to any one of SEQ ID NO: 1-5993: andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in anamount effective to reduce methane production and emission in a ruminant administered the supplement, as compared to a ruminant not administered the supplement.

117. A ruminant supplement capable of reducing methane production and emission in a ruminant, comprising:a) a purified population of bacteria selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(iii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about2026204768   19 Jun 202697% identical to SEQ ID NO: 13; andb) a carrier suitable for ruminant administration;wherein the purified population of bacteria of a) is present in the supplement in anamount effective to reduce methane production and emission in a ruminant administered the supplement,as compared to a ruminant not administered the supplement.

118. The ruminant supplement of claim 104, wherein at least one of the bacteria are selected from:(i) Succinivibrio bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:75,(ii) Prevotella bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO:86, and / or(lii) Bacteroides bacteria with a 16S nucleic acid sequence that is at least about 97% identical to SEQ ID NO: 13.

119. The method of any one of claims 16-18, 50-52, 84-86, or 107-109, wherein the microbes are administered with a prebiotic, a vitamin, or a mineral.

120. The method of any one of claims 16-18, 50-52, 84-86, or 107-109, wherein the microbes are administered with vitamin B or a precursor thereof.

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