Use of lactic acid bacteria to inhibit the growth of methanogens and reduce methane emissions

Abstract

The present invention relates to the use of lactic acid bacteria strains for improving body weight or body composition of animals, such as ruminants and / or monogastric animals, for improving feed efficiency, growth, productivity, and / or milk or meat yield, and / or for inhibiting the growth of methanogenic bacteria and / or archaea in the animal's digestive tract, for reducing the methane production capacity of the rumen and / or gastrointestinal microflora, for reducing methane emissions by animals, for delivering microorganisms to animals, and / or for reducing the greenhouse gas emission footprint of animals. Animal feed compositions are also provided.

Description

[Technical field]

[0001] The present invention relates to the use of lactic acid bacteria strains for improving body weight or body composition of animals, such as ruminants and / or monogastric animals, for improving feed efficiency, growth, productivity, and / or milk or meat yield, and / or for inhibiting the growth of methanogenic bacteria and / or archaea in the animal's digestive tract, for reducing the methane production capacity of the rumen and / or gastrointestinal microflora, for reducing methane emissions by animals, for delivering microorganisms to animals, and / or for reducing the greenhouse gas emission footprint of animals. Animal feed compositions are also provided. [Background technology]

[0002] The use of lactic acid bacteria (LAB) as probiotics to attempt to improve animal health and nutrition is well documented and is being explored as an alternative to antibiotics used as growth promoters.

[0003] On farms, LAB are used as direct-fed microorganisms (DFM), probiotics, and silage inoculants. Their action is exerted in a strain- and host-specific manner. Studies have reported benefits including reduced incidence of diarrhea, enhanced ruminal development, improved feed efficiency, increased weight gain, and reduced morbidity (Krehbiel et al., 2003).

[0004] The use of LAB to reduce methane emissions from ruminant animals has also been proposed.

[0005] The main source of methane emissions is the fermentation of organic matter by methanogenic bacteria and archaea. One of the main sources of anthropogenic methane emissions is agriculture, where methane is produced by enteric fermentation in the digestive tract of ruminants and from manure, such as pig manure. These sources accounted for about 30% of global anthropogenic methane emissions in 2017 (Jackson et al., 2020), and methane emissions from pigs accounted for about 10% of methane production from livestock in China (Mi et al., 2019).

[0006] Furthermore, methane production in ruminants and monogastric animals not only results in greenhouse gas emissions but is also energetically wasteful for the animals. It has long been recognized that methane production dramatically impacts the efficiency with which animals convert feed into metabolic energy. This reduction in efficiency is due to methane representing a caloric loss of approximately 5–10% of total caloric intake in ruminants and approximately 0.1–3.3% of digestive energy losses in pigs (Mi et al., 2019). However, to date, there has been limited research on the potential use of LAB to reduce methane emissions.

[0007] Thus, there remains a need for methods and compositions useful for increasing feed efficiency, increasing body weight or improving body composition, enhancing growth and / or productivity, and / or increasing milk or meat production in animals. Methods and compositions for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal, reducing the ability of the rumen and / or gastrointestinal microflora to produce methane, reducing methane emissions by the animal, delivering microorganisms to the animal, and / or reducing the greenhouse gas emission footprint of the animal are also desirable.

[0008] It is an object of the present invention to go toward achieving one or more of these needs, or at least to provide the public with a useful choice. Summary of the Invention

[0009] In a first aspect, the present invention provides an isolated Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0010] In one embodiment, the Lacticaseibacillus rhamnosus strain FNZ129 is a biologically pure culture.

[0011] In a second aspect, the present invention provides a food or feed composition comprising Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated 2 August 2021) or a derivative thereof.

[0012] In a third aspect, the present invention provides a method for producing a composition comprising the steps of: a) improving the body weight and / or body composition of an animal; b) Increases the feed efficiency of animals; c) enhancing animal growth and / or productivity; d) increasing the yield of milk and / or milk components produced by the animal; e) inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of animals; f) reducing the ability of the gastrointestinal microflora to produce methane; g) Reduce meth excretion from animals; h) delivering the animal to the microorganism; and / or i) A feed composition for reducing greenhouse gas emissions in animals is provided, the feed composition comprising Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0013] In a fourth aspect, the present invention provides a method for improving the body weight and / or body composition of an animal, said method comprising the step of administering to said animal a food or feed composition of the second aspect, or a feed composition of the third aspect.

[0014] In a fifth aspect, the present invention provides a method of increasing feed efficiency in an animal, said method comprising administering to said animal a food or feed composition of the second aspect, or a feed composition of the third aspect.

[0015] In a sixth aspect, the present invention provides a method for enhancing growth and / or productivity in an animal, said method comprising administering to said animal a food or feed composition of the second aspect, or a feed composition of the third aspect.

[0016] In a seventh aspect, the present invention provides a method of increasing the yield of milk and / or milk components produced from an animal, said method comprising administering to said animal a food or feed composition of the second aspect, or a feed composition of the third aspect.

[0017] In an eighth aspect, the present invention provides a method for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal, said method comprising administering to said animal a food or feed composition of the second aspect or a feed composition of the third aspect.

[0018] In a ninth aspect, the present invention provides a method for reducing the methane producing potential of the gastrointestinal microflora of an animal, said method comprising administering to said animal a food or feed composition of the second aspect, or a feed composition of the third aspect.

[0019] In a tenth aspect, the present invention provides a method for reducing methane emissions by an animal, said method comprising administering to said animal a food or feed composition of the second aspect, or a feed composition of the third aspect.

[0020] In an eleventh aspect, the present invention provides a method for delivering a microorganism to an animal, said method comprising administering to said animal a food or feed composition of the second aspect, or a feed composition of the third aspect.

[0021] In a twelfth aspect, the present invention provides a method for reducing the greenhouse gas emission footprint of an animal, said method comprising administering to said animal a food or feed composition of the second aspect, or a feed composition of the third aspect.

[0022] In a thirteenth aspect, the present invention provides a method for improving body weight and / or body composition in an animal, said method comprising administering to said animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0023] In a fourteenth aspect, the present invention provides a method of increasing feed efficiency in an animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0024] In a fifteenth aspect, the present invention provides a method for enhancing growth and / or productivity in an animal, said method comprising the step of administering to said animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0025] In a sixteenth aspect, the present invention provides a method of increasing the yield of milk and / or milk components produced from an animal, said method comprising the step of administering to said animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0026] In a seventeenth aspect, the present invention provides a method for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0027] In an eighteenth aspect, the present invention provides a method for reducing the ability of gastrointestinal microflora to produce methane, the method comprising administering to an animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0028] In a nineteenth aspect, the present invention provides a method for reducing methane emissions by an animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0029] In a twentieth aspect, the present invention provides a method for delivering a microorganism to an animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0030] In a twenty-first aspect, the present invention provides a method for reducing the greenhouse gas emission footprint of an animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0031] In a twenty-second aspect, the present invention provides an animal to which the method of any one of the fourth to twenty-first aspects has been applied.

[0032] In a twenty-third aspect, the present invention provides a method for producing an animal product, such as a dairy product, a meat product, or a wool product, having a reduced greenhouse gas emission footprint, the method comprising: a. providing an animal of the 22nd aspect, and b. Producing animal products from animals Includes.

[0033] In a twenty-fourth aspect, the present invention provides use of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof for the manufacture of a composition for improving body weight and / or body composition of an animal, increasing feed efficiency in an animal, increasing growth and / or productivity in an animal, increasing the yield of milk and / or milk components produced from an animal, inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal, reducing the ability of the gastrointestinal microflora to produce methane, reducing methane emissions by an animal, delivering microorganisms to an animal, and / or reducing the greenhouse gas emission footprint of an animal.

[0034] In a twenty-fifth aspect, the present invention provides Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof for use in improving the body weight and / or body composition of an animal, increasing the feed efficiency of an animal, enhancing the growth and / or productivity of an animal, increasing the yield of milk and / or milk components produced from an animal, inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal, reducing the ability of the gastrointestinal microflora to produce methane, reducing methane emissions by an animal, delivering microorganisms to an animal, and / or reducing the greenhouse gas emission footprint of an animal.

[0035] In a twenty-sixth aspect, the present invention provides a ruminant feed composition for inhibiting the growth of methanogenic bacteria and / or archaea in the forestomach of a ruminant, reducing the ability of the rumen microflora to produce methane, reducing methane emissions by a ruminant, increasing feed efficiency in a ruminant, increasing growth and / or productivity in a ruminant, increasing the yield of milk and / or milk components produced from the ruminant, or improving the body weight and / or body composition of a ruminant, the feed composition comprising Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated 2 August 2021) or a derivative thereof.

[0036] In a twenty-seventh aspect, the present invention provides a method for inhibiting the growth of methanogenic bacteria and / or archaea in the forestomach of a ruminant animal, said method comprising the step of administering to said animal a food or feed composition according to the second aspect, or a ruminant feed composition according to the twentieth aspect.

[0037] In a twenty-eighth aspect, the present invention provides a method for reducing the ruminal microbiota potential of a ruminant comprising administering to said animal a food or feed composition according to the second aspect, or a ruminant feed composition according to the twentieth aspect.

[0038] In a twenty-ninth aspect, the present invention provides a method for reducing methane emission by a ruminant animal, said method comprising administering to said animal a food or feed composition according to the second aspect, or a ruminant feed composition according to the twentieth aspect.

[0039] In a thirtieth aspect, the present invention provides a method for increasing feed efficiency in a ruminant animal, said method comprising administering to said animal a food or feed composition according to the second aspect, or a ruminant feed composition according to the twentieth aspect.

[0040] In a thirty-first aspect, the present invention provides a method of increasing growth and / or productivity in a ruminant animal, said method comprising administering to said animal a food or feed composition according to the second aspect, or a ruminant feed composition according to the twentieth aspect.

[0041] In a thirty-second aspect, the present invention provides a method of increasing the yield of milk and / or milk components produced from a ruminant animal, said method comprising administering to said animal a food or feed composition according to the second aspect, or a ruminant feed composition according to the twentieth aspect.

[0042] In a thirty-third aspect, the present invention provides a method of improving the body weight or body composition of a ruminant, said method comprising the step of administering to said animal a food or feed composition according to the second aspect, or a ruminant feed composition according to the twentieth aspect.

[0043] In a thirty-fourth aspect, the present invention provides a method for inhibiting the growth of methanogenic bacteria and / or archaea in the forestomach of a ruminant comprising administering to the ruminant an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0044] In a thirty-fifth aspect, the present invention provides a method for reducing methane emission by a ruminant animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0045] In a thirty-sixth aspect, the present invention provides a method of increasing feed efficiency in a ruminant animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0046] In a thirty-seventh aspect, the present invention provides a method for enhancing growth and / or productivity in a ruminant animal, said method comprising the step of administering to the animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0047] In a thirty-eighth aspect, the present invention provides a method for increasing the yield of milk and / or milk components produced from a ruminant animal, said method comprising the step of administering to the animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0048] In a thirty-ninth aspect, the present invention provides a method of improving body weight and / or body composition of a ruminant animal, said method comprising the step of administering to said animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0049] In a fortieth aspect, the present invention provides a method for reducing the ability of ruminal microflora to produce methane, the method comprising administering to an animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0050] In a forty-first aspect, the present invention provides a method for improving the absorptive capacity of the forestomach, e.g., for increasing the absorptive capacity of volatile fatty acids (VFAs), comprising administering to an animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0051] In a forty-second aspect, the present invention provides a method for enhancing the physical and / or functional development of the rumen, or other cavity of the forestomach, in a ruminant, such as a young ruminant, such as a pre-weaned young ruminant, comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0052] In a forty-third aspect, the present invention provides the use of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated 2 August 2021) or a derivative thereof for the manufacture of a composition for inhibiting the growth of methanogenic bacteria and / or archaea in the forestomach of a ruminant, reducing the ability of the ruminal microflora to produce methane, reducing methane emissions by a ruminant, increasing feed efficiency in a ruminant, increasing the yield of milk and / or milk components produced from a ruminant, or improving the body weight and / or body composition of a ruminant.

[0053] In a forty-fourth aspect, the present invention provides Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated 2 August 2021) or a derivative thereof for use in inhibiting the growth of methanogenic bacteria and / or archaea in the forestomach of a ruminant, reducing the ability of the ruminal microflora to produce methane, reducing methane emissions by a ruminant, increasing feed efficiency in a ruminant, increasing the yield of milk and / or milk components produced from a ruminant, or improving the body weight and / or body composition of a ruminant.

[0054] In a forty-fifth aspect, the present invention provides a monogastric feed composition for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of a monogastric animal, reducing the methane producing capacity of the gastrointestinal microflora, reducing methane emission by a monogastric animal, increasing feed efficiency in a monogastric animal, enhancing growth and / or productivity in a monogastric animal, increasing the yield of milk and / or milk components produced from the monogastric animal or improving the body weight and / or body composition of a monogastric animal, wherein the feed composition comprises Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0055] In a further aspect, the present invention provides a method for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of a monogastric animal, said method comprising the step of administering to said animal a food or feed composition according to the second aspect or a monogastric feed composition according to the forty-fifth aspect.

[0056] In a further aspect, the present invention provides a method for reducing the methane producing potential of the gastrointestinal microflora, said method comprising administering to said animal a food or feed composition according to the second aspect or a monogastric feed composition according to the forty-fifth aspect.

[0057] In a further aspect, the present invention provides a method for reducing methane emission by a monogastric animal, said method comprising administering to said animal a food or feed composition according to the second aspect or a monogastric feed composition according to the forty-fifth aspect.

[0058] In a further aspect, the present invention provides a method of increasing feed efficiency in a monogastric animal, said method comprising the step of administering to said animal a food or feed composition according to the second aspect, or a monogastric feed composition according to the forty-fifth aspect.

[0059] In a further aspect, the present invention provides a method of increasing the growth and / or productivity of a monogastric animal, said method comprising the step of administering to said animal a food or feed composition according to the second aspect, or a monogastric feed composition according to the forty-fifth aspect.

[0060] In a further aspect, the present invention provides a method of increasing the yield of milk and / or milk components produced from a monogastric animal, said method comprising the step of administering to said animal a food or feed composition according to the second aspect, or a monogastric feed composition according to the forty-fifth aspect.

[0061] In a further aspect, the present invention provides a method of improving body weight or body composition of a monogastric animal, said method comprising the step of administering to said animal a food or feed composition according to the second aspect, or a monogastric feed composition according to the forty-fifth aspect.

[0062] In a further aspect, the present invention provides a method for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of a monogastric animal, the method comprising administering to the monogastric animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0063] In a further aspect, the present invention provides a method for reducing methane emission by a monogastric animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0064] In a further aspect, the present invention provides a method of increasing feed efficiency in a monogastric animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0065] In a further aspect, the present invention provides a method for reducing the ability of gastrointestinal microflora to produce methane, the method comprising administering to an animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0066] In a further aspect, the present invention provides a method for enhancing growth and / or productivity in a monogastric animal, the method comprising administering an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof to the monogastric animal.

[0067] In a further aspect, the present invention provides a method for improving body weight and / or body composition of a monogastric animal, the method comprising administering an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof to the monogastric animal.

[0068] In a further aspect, the present invention provides a method for improving the absorptive capacity of the gastrointestinal tract, for example for increasing the absorptive capacity of volatile fatty acids (VFAs), comprising administering to an animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0069] In a further aspect, the present invention provides a method for promoting physical and / or functional development of the gastrointestinal tract, functional achievement of the gastrointestinal tract, or maturation of the gastrointestinal tract in a monogastric animal, such as a young animal, such as a pre-weaned young animal, comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0070] In a further aspect, the present invention provides the use of Lacticaseibacillus rhamnosus strain FNZ129, NMIA accession number V21 / 015446 dated August 2, 2021, or a derivative thereof, for the manufacture of a composition for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of a monogastric animal, reducing the methane producing capacity of the gastrointestinal microflora, reducing methane production by a monogastric animal, increasing feed efficiency in a monogastric animal, or improving the body weight and / or body composition of a monogastric animal.

[0071] In a further aspect, the present invention provides Lacticaseibacillus rhamnosus strain FNZ129, NMIA accession number V21 / 015446 dated 2 August 2021, or a derivative thereof, for use in inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of a monogastric animal, reducing the methane producing capacity of the gastrointestinal microflora by a monogastric animal, reducing methane production, increasing feed efficiency in a monogastric animal, or improving body weight and / or body composition in a monogastric animal.

[0072] In a further aspect, the present invention provides a method for delivering a microorganism to an animal, said method comprising administering to said animal a food or feed composition according to the second aspect, or a feed composition according to the third aspect.

[0073] In a further aspect, the present invention provides a method for delivering a microorganism to an animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0074] The following embodiments and preferences may relate to any of the above aspects, either alone or in any combination of any two or more.

[0075] In one embodiment, the isolated Lacticaseibacillus rhamnosus strain FNZ129 is a biologically pure culture.

[0076] In some embodiments, the food or feed composition is a ruminant feed composition. In some embodiments, the feed composition is a ruminant feed composition and the animal is a ruminant. In some embodiments, the food or feed composition is a feed composition for a monogastric animal. In some embodiments, the feed composition is a monogastric feed composition and the animal is a monogastric animal.

[0077] In some embodiments, the feed composition is a fermented yogurt-style composition, which is formed through a process of growing L. rhamnosus FNZ129 using a milk-based carrier or a non-dairy-based carrier.

[0078] In some embodiments, the feed composition is or comprises a partial or complete mixed ration (TMR), corn, soybean, forage, cereals, distillers grains, germinated grains, legumes, fiber, forage, grass, hay, straw, silage, grain, leaves, meal, mash feed, fruit pulp, vegetable pulp, fruit or vegetable pomace, citrus meal, wheat shorts, corn cob meal, lick block, or molasses.

[0079] In some embodiments, the feed composition further comprises at least one microorganism of a different species or strain, a methanogen or a vaccine that inhibits methane production, and / or a natural or chemically synthesized methane production inhibitor, and / or a methane production inhibitor such as bromoform.

[0080] In some embodiments, the feed composition further comprises one or more agents selected from one or more prebiotics, one or more probiotics, one or more postbiotics, one or more sources of dietary fiber, one or more galactooligosaccharides, one or more short chain galactooligosaccharides, one or more long chain galactooligosaccharides, one or more fructooligosaccharides, inulin, one or more galactans, one or more fructans, lactulose, one or more milk-derived oligosaccharides (e.g., 2'-fucosyllactose, 3'-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, lacto-N-tetraose, lacto-N-neotetraose), or any mixture of any two or more thereof.

[0081] In some embodiments, the derivative of L. rhamnosus FNZ129 is a cell lysate of L. rhamnosus FNZ129, a cell suspension of L. rhamnosus FNZ129, a metabolic product of L. rhamnosus FNZ129, or a culture supernatant of L. rhamnosus FNZ129 or killed L. rhamnosus FNZ129.

[0082] In some embodiments, the ruminant feed composition is a cattle feed composition. In some embodiments, the ruminant feed composition is a goat feed composition. In some embodiments, the ruminant feed composition is a sheep feed composition. In some embodiments, the ruminant feed composition is a bison feed composition. In some embodiments, the ruminant feed composition is a yak feed composition. In some embodiments, the ruminant feed composition is a buffalo feed composition. In some embodiments, the ruminant feed composition is a deer feed composition. In some embodiments, the ruminant feed composition is a camel feed composition. In some embodiments, the ruminant feed composition is an alpaca feed composition. In some embodiments, the ruminant feed composition is a llama feed composition. In some embodiments, the ruminant feed composition is a wildebeest feed composition. In some embodiments, the ruminant feed composition is an antelope feed composition. In some embodiments, the ruminant feed composition is a nilgai feed composition.

[0083] In some embodiments, the L. rhamnosus FNZ129 or derivative thereof is administered in a feed composition. In some embodiments, the feed composition is a livestock feed composition. In some embodiments, the feed composition is a companion animal feed composition. In some embodiments, the feed composition is a pet food composition. In some embodiments, the feed composition is a herbivore feed composition. In some embodiments, the feed composition is an omnivorous feed composition. In some embodiments, the feed composition is a carnivore feed composition. In some embodiments, the feed composition is a poultry feed composition. In some embodiments, the feed composition is a cat feed composition. In some embodiments, the feed composition is a dog feed composition. In some embodiments, the feed composition is a pig feed composition. In some embodiments, the feed composition is a horse feed composition. In some embodiments, the feed composition is a donkey feed composition. In some embodiments, the feed composition is a rabbit feed composition. In some embodiments, the feed composition is a chicken feed composition. In some embodiments, the feed composition is a duck feed composition. In some embodiments, the feed composition is a goose feed composition. In some embodiments, the feed composition is a turkey feed composition. In some embodiments, the feed composition is a human food composition.

[0084] In some embodiments, the animal is a ruminant. In some embodiments, the animal is a monogastric animal.

[0085] In one embodiment, the method improves ruminal anatomical development, e.g., the method enhances ruminal epithelial development and / or muscularization, e.g., ruminal mass growth, ruminal papilla growth, increased papilla density, e.g., dorsal papilla density, and / or total ruminal wall surface area in an animal.

[0086] In one embodiment, the method increases, for example, rumen weight, rumen wall thickness, or ruminal papilla density per centimeter2 of ruminal wall compared to untreated animals.

[0087] In one embodiment, the method increases the length, width, and / or surface area of ​​the ruminal papillae. For example, in some embodiments, the method increases the length, width, and / or surface area of ​​the ruminal papillae by at least 1.01, 1.02, 1.03, 1.04, 1.05, 1.06, 1.07, 1.08, 1.09, 1.10, 1.11, 1.12, 1.13, 1.14, 1.15, 1.16, 1.17, 1.18, 1.19, 1.20, 1.22, 1.24, 1.26, 1.28, 1.30, 1.32, 1.34, 1.36, 1.38, or 1.40 times that of an untreated animal.

[0088] In one embodiment, the method enhances ruminal functional performance or promotes forestomach maturation, for example, the method stimulates rumination, increases dry matter intake (DMI), increases absorptive capacity, and / or promotes maturation to mature physiology.

[0089] In some embodiments, the method inhibits the growth of methylotrophic methanogens in the forestomach and / or cecum of the animal. In some embodiments, the method inhibits the growth of methanogens from the genus Methanosphaera in the forestomach and / or cecum of the animal. In some embodiments, the method inhibits the growth of methylotrophic methanogens in the gastrointestinal tract of the animal. In one embodiment, the method inhibits the growth of methanogens from the genus Methanosphaera in the gastrointestinal tract of the animal. In some embodiments, the method inhibits the growth of methylotrophic methanogens in the cecum or colorectum of the animal. In one embodiment, the method inhibits the growth of methanogens from the genus Methanosphaera in the cecum or colorectum of the animal.

[0090] In some embodiments, L. rhamnosus FNZ129 or a derivative thereof is administered in a composition that is a food, beverage, food additive, beverage additive, animal feed, animal feed additive, animal feed supplement, dietary supplement, carrier, vitamin or mineral premix, nutritional product, enteral nutritional product, soluble, slurry, supplement, pharmaceutical, lick block, drench, tablet, capsule, pellet, bolus, or intraruminal product, or L. rhamnosus FNZ129 is encapsulated in, for example, a liposome, microbubble, microparticle, or microcapsule.

[0091] In a further aspect, the invention provides a composition comprising Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. In some embodiments, the composition is a food, beverage, food additive, beverage additive, animal feed, animal feed additive, animal feed supplement, dietary supplement, carrier, vitamin or mineral premix, nutritional product, enteral nutritional product, soluble, slurry, supplement, pharmaceutical, lick block, drench, tablet, capsule, pellet, bolus, or intraruminal product, or L. rhamnosus FNZ129 is encapsulated, for example, in a liposome, microbubble, microparticle, or microcapsule.

[0092] In some embodiments, L. rhamnosus FNZ129 or a derivative thereof is administered in drinking water, milk, milk powder, milk replacer, milk fortifier, whey, whey powder, partial or total mixed ration (TMR), feed pellets, corn, soybeans, feed, cereals, distillers grains, germinated grains, legumes, vitamins, amino acids, minerals, fiber, forage, grass, hay, straw, silage, grains, leaves, meal, solubles, slurry, supplements, mash feed, meal, fruit pulp, vegetable pulp, fruit or vegetable pomace, citrus meal, wheat shorts, corn cob meal, molasses, sucrose, maltodextrin, rice hulls, vermiculite, zeolite, or ground limestone.

[0093] The method comprises administering to the animal at least about 10 4 Colony forming units / kg dry weight carrier feed, e.g., at least about 10 5 , at least about 10 6 , at least about 10 7 , at least about 10 8 , at least about 10 9 , at least about 10 10 , at least about 10 11 , at least about 10 12 , or at least about 10 13 In some embodiments, the method comprises administering to the animal 10 colony forming units / kg dry weight of carrier feed of L. rhamnosus FNZ118. 4 ~10 13 In one embodiment, the method comprises administering to the animal 10 colony forming units of L. rhamnosus FNZ129 per kg dry weight of carrier feed. 8 ~10 12 The method comprises administering to the animal at least one colony forming unit of L. rhamnosus FNZ129.

[0094] In some embodiments, the method comprises administering 10 mg / kg of animal body weight. 4 ~10 10 The method includes administering to the animal at least one colony forming unit / day of the antibody.

[0095] In some embodiments, the method comprises culturing L. rhamnosus FNZ129 for 10 4 ~10 13 In some embodiments, the method comprises administering L. rhamnosus FNZ129 in an amount of 10 8 ~10 12 The method includes administering 1000 colony forming units / day.

[0096] In some embodiments, the method further comprises administering at least one microorganism of a different species or strain, a methanogen or a vaccine that inhibits methane production, and / or a natural or chemically synthesized methane production inhibitor and / or a methane production inhibitor. An example of a useful inhibitor of methane production is bromoform, which acts by inhibiting the efficiency of the methyltransferase enzyme by reacting with the reduced vitamin B12 cofactor required for the final stage of methane production.

[0097] In one embodiment, the method further comprises administering at least one microorganism of a different species or strain, a methanogen or a vaccine that inhibits methane production, and / or a natural or chemically synthetic methane production inhibitor, and / or a methane production inhibitor that targets hydrogenotrophic methanogens, e.g., methanogens from the genus Methanobrevibacter.

[0098] In some embodiments, L. rhamnosus FNZ129 or a derivative thereof is administered separately, simultaneously, or sequentially (sequentially) with one or more agents selected from one or more prebiotics, one or more probiotics, one or more postbiotics, one or more dietary fiber sources, one or more galactooligosaccharides, one or more short chain galactooligosaccharides, one or more long chain galactooligosaccharides, one or more fructooligosaccharides, inulin, one or more galactans, one or more fructans, lactulose, or any mixture of any two or more thereof.

[0099] In some embodiments, the methods further enhance the growth or productivity of the animal, for example, the methods increase the yield of milk and / or milk components produced from the animal, hi some embodiments, the methods increase the yield of milk fat, milk protein, or milk solids in the milk produced from the animal.

[0100] In some embodiments, the method further increases the body weight and / or improves the body composition (e.g., changes the muscle to fat ratio) of the animal. In some embodiments, the method increases the body weight and / or improves the body composition (e.g., changes the muscle to fat ratio) of the monogastric animal. In some embodiments, the method reduces the body mass index (BMI) and / or increases the muscle to fat ratio of the monogastric animal.

[0101] In some embodiments, the ruminant is a cow, goat, sheep, bison, yak, buffalo, deer, camel, alpaca, llama, wildebeest, antelope, or nilgai. In one embodiment, the ruminant is a cow or a sheep. In one embodiment, the ruminant is a cow. In one embodiment, the ruminant is a lactating animal. In an alternative embodiment, the ruminant is a pre-weaned animal, such as a calf or lamb.

[0102] In some embodiments, the methods further increase wool growth in ruminant animals.

[0103] In some embodiments, the monogastric animal is a human, a pig, a cat, a dog, a horse, a donkey, a rabbit, or a poultry. In some embodiments, the monogastric animal is a companion animal. In some embodiments, the monogastric animal is a non-human animal. In some embodiments, the monogastric animal is a pig. In some embodiments, the monogastric animal is a chicken, a duck, a goose, or a turkey.

[0104] In some embodiments, the animal is a pre-weaned animal, for example, a calf, lamb, piglet, or foal.

[0105] In some embodiments, L. rhamnosus FNZ129 is administered to the animal both pre-weaning and post-weaning.

[0106] In some embodiments, administration is performed on a pre-weaned animal, and the inhibition of the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of the animal, reduction in methane emissions, e.g., methane production, by the animal, and / or increased feed efficiency in the animal persists after weaning.

[0107] In some embodiments, the inhibition of growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of the animal, the reduction in methane emissions by the animal, e.g., methane production, and / or the increase in feed efficiency in the animal persists for at least 2 days, 3 days, 5 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, or 7 years following the final administration of L. rhamnosus FNZ129.

[0108] In some embodiments, the inhibition of the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of the animal, the reduction in methane emissions, e.g., methane production, by the animal, and / or the increase in feed efficiency in the animal persists for the life of the animal.

[0109] In some embodiments the composition is or comprises a food or feed composition according to the second aspect, or a feed composition according to the third aspect.

[0110] The invention may also be broadly described as consisting of any and all combinations of the parts, elements and features referred to or shown in the specification of this application, either individually or collectively, and any two or more of said parts, elements or features, and where a particular integer having a known equivalent in the art to which this invention pertains is referred to herein, such known equivalent is deemed to be incorporated herein as if individually set forth.

[0111] Reference to a range of numbers disclosed herein (e.g., 1-10) is intended to incorporate reference to all rational numbers within that range (e.g., 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9, and 10) and also any range of rational numbers within that range (e.g., 2-8, 1.5-5.5, and 3.1-4.7), and thus all subranges of every range expressly disclosed herein are hereby expressly disclosed. These are merely examples of what is specifically intended, and all possible combinations of numerical values ​​between the lowest and highest values ​​recited should be considered to be expressly set forth in this application in a similar manner.

[0112] The term "comprising" as used in this specification means "consisting at least in part of." When interpreting each statement in this specification that contains the term "comprising," there may be other features present than the feature prefaced by that term. Related terms such as "comprise" and "comprises" should be interpreted in the same manner.

[0113] Where this specification refers to patents, other external documents, or other sources of information, it is generally for the purpose of providing a context for discussing features of the present invention. Unless specifically stated otherwise, a reference to such an external document is not to be construed as an admission that such document or such source is prior art or part of the common general knowledge in the art in any jurisdiction. [Brief description of the drawings]

[0114] [Figure 1] FIG. 1 shows the effect of dietary supplementation with L. rhamnosus FNZ129 on pig growth. [Diagram 2] Figure 2 shows the effect of L. rhamnosus FNZ129 on the most probable number (MPN) of methanogens per gram of pig feces in the cecum. L. rhamnosus significantly reduced the MPN (p=0.015). [Diagram 3]FIG. 3 shows the effect of L. rhamnosus FNZ129 on the most probable number (MPN) of methanogens per gram of porcine feces in the large intestine. [Figure 4] Figure 4 shows absolute values ​​of the main volatile fatty acids measured in cecal samples taken from pigs after euthanasia. n=8 in the control group and n=7 in the FNZ129-treated group. *T-test FNZ129 vs. control p<0.05. [Diagram 5] Figure 5 shows absolute values ​​of the main volatile fatty acids measured in colorectal samples taken from pigs after euthanasia: n=8 in the control group and n=7 in the FNZ129-treated group. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0115] The present invention is based on the finding that Lacticaseibacillus rhamnosus FNZ129 strain and its derivatives inhibit or suppress the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of animals and / or reduce the methane production capacity of the gastrointestinal microflora. For example, L. rhamnosus FNZ129 and its derivatives inhibit or suppress the growth of methanogenic bacteria and / or archaea in the forestomach of ruminants and / or reduce the methane production capacity of the ruminal microflora. L. rhamnosus FNZ129 and its derivatives also inhibit the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of monogastric animals and / or reduce the methane production capacity of the ruminal microflora. Inhibiting the growth of methanogenic bacteria and / or archaea can reduce methane emissions and can alter the volatile fatty acid (VFA) profile, total VFA concentration, residual feed intake (RFI) and / or fermentation rates in the gastrointestinal tract, which can act as an increased energy source to drive enhanced growth or increased productivity, such as milk, meat, or wool production, and can stimulate gastrointestinal development, such as rumen and / or ruminal papilla development.

[0116] Thus, in a first aspect, the present invention provides an isolated Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0117] In a second aspect, the present invention provides a food or feed composition comprising Lacticaseibacillus rhamnosus strain FNZ129 (NMIA accession number V21 / 015446 dated 2 August 2021) or a derivative thereof.

[0118] In a third aspect, the present invention provides a method for producing a composition comprising the steps of: a) improving the body weight and / or body composition of an animal; b) Increases the feed efficiency of animals; c) enhancing animal growth and / or productivity; d) increasing the yield of milk and / or milk components produced by the animal; e) inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of animals; f) reducing the ability of the gastrointestinal microflora to produce methane; g) Reduce meth excretion from animals; h) delivering the animal to the microorganism; and / or i) A feed composition for reducing greenhouse gas emissions in animals is provided, the feed composition comprising Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0119] In a further aspect, the present invention provides a method for improving the body weight and / or body composition of an animal, said method comprising the step of administering to said animal a food or feed composition of the second aspect or a feed composition of the third aspect.

[0120] In a further aspect, the present invention provides a method of increasing feed efficiency in an animal, said method comprising administering to said animal a food or feed composition of the second aspect, or a feed composition of the third aspect.

[0121] In a further aspect, the present invention provides a method for enhancing growth and / or productivity in an animal, said method comprising administering to said animal a food or feed composition of the second aspect or a feed composition of the third aspect.

[0122] In a further aspect, the present invention provides a method of increasing the yield of milk and / or milk components produced from an animal, said method comprising administering to said animal a food or feed composition of the second aspect, or a feed composition of the third aspect.

[0123] In a further aspect, the present invention provides a method for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal, said method comprising administering to said animal a food or feed composition of the second aspect or a feed composition of the third aspect.

[0124] In a further aspect, the present invention provides a method for reducing the methane producing potential of the gastrointestinal microflora of an animal, said method comprising administering to said animal a food or feed composition of the second aspect or a feed composition of the third aspect.

[0125] In a further aspect, the present invention provides a method for reducing methane emissions by an animal, said method comprising administering to said animal a food or feed composition of the second aspect or a feed composition of the third aspect.

[0126] In a further aspect, the present invention provides a method for delivering a microorganism to an animal, said method comprising administering to said animal a food or feed composition of the second aspect or a feed composition of the third aspect.

[0127] In a further aspect, the present invention provides a method for reducing the greenhouse gas emission footprint of an animal, said method comprising administering to said animal a food or feed composition of the second aspect, or a feed composition of the third aspect.

[0128] In a further aspect, the present invention provides a method for improving body weight and / or body composition in an animal, said method comprising administering to said animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0129] In a further aspect, the present invention provides a method for increasing feed efficiency in an animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0130] In a further aspect, the present invention provides a method for enhancing growth and / or productivity in an animal, said method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0131] In a further aspect, the present invention provides a method of increasing the yield of milk and / or milk components produced from an animal, said method comprising the step of administering to said animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0132] In a further aspect, the present invention provides a method for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0133] In a further aspect, the present invention provides a method for reducing the ability of gastrointestinal microflora to produce methane, the method comprising administering to an animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0134] In a further aspect, the present invention provides a method for reducing methane emissions by an animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0135] In a further aspect, the present invention provides a method for delivering a microorganism to an animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0136] In a further aspect, the present invention provides a method for reducing the greenhouse gas emission footprint of an animal, the method comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0137] In a further aspect, there is provided a use of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof for the manufacture of a composition for improving the body weight and / or body composition of an animal, increasing feed efficiency in an animal, increasing growth and / or productivity in an animal, increasing the yield of milk and / or milk components produced from an animal, inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal, reducing the ability of the gastrointestinal microflora to produce methane, reducing methane emissions by an animal, delivering microorganisms to an animal, and / or reducing the greenhouse gas emission footprint of an animal.

[0138] In a further aspect, the present invention provides Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof for use in improving the body weight and / or body composition of an animal, increasing the feed efficiency of an animal, enhancing the growth and / or productivity of an animal, increasing the yield of milk and / or milk components produced from an animal, inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal, reducing the ability of the gastrointestinal microflora to produce methane, reducing methane emissions by an animal, delivering microorganisms to an animal, and / or reducing the greenhouse gas emission footprint of an animal.

[0139] In one embodiment, the methods and compositions enhance ruminal anatomical development, for example, the methods enhance ruminal epithelial development and / or muscularization, e.g., ruminal mass growth, ruminal papilla growth, increased papilla density, e.g., dorsal papilla density, and / or total ruminal wall surface area in an animal.

[0140] In one embodiment, the methods and compositions disclosed herein increase rumen weight, rumen wall thickness, or ruminal papilla density per cm2 of ruminal wall.

[0141] In one embodiment, the methods and compositions disclosed herein increase the functional performance of the rumen. For example, the methods stimulate rumination and / or increase dry matter intake (DMI). In one embodiment, the methods and compositions disclosed herein increase ruminal turnover rate and / or increase post-ruminal digestion. Without wishing to be bound by theory, it is hypothesized that a higher ruminal turnover rate selects for microorganisms capable of rapid heterofermentative growth on soluble sugars, which produces less hydrogen and leads to less methane formation. For example, Kamke et al. (2016) note that conversion of lactate to butyrate rather than propionate produces 2 moles of hydrogen per hexose, which can produce 0.5 moles of methane via the hydrogenotrophic pathway, and hypothesize that direct fermentation of hexose to butyrate and acetate by members of the Ruminococcaceae produces 2.66 moles of hydrogen and allows for the formation of 0.66 moles of methane. Therefore, lower hydrogen production via the lactate to butyrate pathway is predicted to reduce methane production.

[0142] The term "administering" refers to the act of introducing an effective amount of the L. rhamnosus FNZ129 strain or a derivative thereof into the forestomach of a ruminant animal. More particularly, this administration is by the oral route. This administration can be carried out in particular by supplementing the strain in animal feed or drink and allowing the animal to ingest the supplemented feed or drink.

[0143] The term "effective amount" refers to an amount of L. rhamnosus strain FNZ129 or a derivative thereof sufficient to achieve the desired effect, i.e., inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal (e.g., the forestomach of an animal), reducing methane emission by the animal, or increasing feed efficiency in an animal, as compared to a reference. The desired effect (e.g., inhibiting the growth of methanogenic bacteria and / or archaea and / or reducing methane production or emission) can be measured in vitro or in vivo. For example, the desired effect can be measured in vitro by oral administration to an animal, e.g., in an artificial ruminal system such as that described in the Examples below or in T. Hano (1993) J. Gen. Appl. Microbiol., 39, 35-45, or using the methods described herein.

[0144] This effective amount can be administered to the animal in one or more doses.

[0145] The terms "reducing methane production" and "reducing methane emissions", e.g., "reducing methane production by animals" and "reducing methane emissions by animals", refer to reducing methane production or emissions by any mechanism and from any animal-related source. For example, when referring to ruminant animals, the terms may refer to a reduction in methane produced in the forestomach of the ruminant animal, or a reduction in methane produced or released by the feces of the ruminant animal. When referring to monogastric animals, the terms may refer to a reduction in methane produced in the gastrointestinal tract of the monogastric animal, or a reduction in methane produced or released by the feces or manure of the monogastric animal.

[0146] The reduction in methane production is expected to be due to a variety of mechanisms, including, for example, killing methanogens (i.e., a bactericidal / archaecidal effect), inhibiting the growth of methanogens (i.e., a bacteriostatic / archaecidal effect), and / or inhibiting the ability of the gastrointestinal, forestomach, or ruminal microbiota to produce methane. Inhibiting the ability of the gastrointestinal, forestomach, or ruminal microbiota to produce methane may be through a variety of mechanisms, including, for example, physical and / or chemical changes to the gastrointestinal tract, cecum, forestomach, or ruminal environment, changes to the microbiota, inhibition of one or more methane production pathways, and / or intermediate cross-feeding (or prevention of cross-feeding) between members of the microbiota.

[0147] It is understood that reducing greenhouse gas (GHG) emissions, such as methane emissions, is desirable. GHG emissions can be reduced directly or indirectly, for example, by reducing the ability of the gastrointestinal microflora to produce methane and / or by reducing methane emissions by animals. One example of an indirect reduction of GHG emissions is through land use changes or land disposal. Animals with improved feed efficiency (e.g., animals to which the method or composition of the present invention has been applied) require less pasture for roughage and / or less imported feed. Alternatively, or in addition, more animals can be raised on a given land area, allowing for the same production with reduced land use. In any case, the reduction in land requirements may allow unused pasture to be retired, for example by planting trees or other vegetation for carbon sequestration. Such land use changes may further reduce GHG emissions per farm, resulting in lower GHG emissions per animal and / or per kg of animal product (dairy products, meat, wool products, etc.).

[0148] The GHG emissions of animals and / or animal products can be determined using techniques known in the art. It is recognized that certain GHGs produce more global warming potential than others. For example, emitting 1 kg of methane gas produces a global warming effect equivalent to 25 kg of CO2. To account for this, GHG emissions are typically reported as CO2 equivalents (CO2e), which is the amount of CO2 equivalent to global warming. GHG emission footprints can be calculated per animal or per amount of animal product (e.g., per kg of milk solids, per kg of meat, per kg of wool). As mentioned above, the GHG emission footprint should take into account land use changes, such as planting trees and other vegetation for carbon sequestration.

[0149] The term "animal product" refers to any product produced from or by animals, or that contains animal-derived ingredients. This term is intended to include products produced directly by animals (e.g., milk, meat, and wool), as well as products that contain animal ingredients or are produced from animal ingredients, optionally with other ingredients, and optionally further processed. For example, this term is intended to include foods and beverages that contain animal ingredients, such as various dairy products (including buttermilk, cheese, cream, formula, ice cream, milk, milk powder, puddings, shakes, smoothies, and yogurt), meat products (such as chops, minced meat, hamburgers, sausages, sausage meat, steaks, and chicken wings), and other products that contain animal ingredients.

[0150] The term "feed efficiency" refers to the relationship between feed intake and muscle gain or milk production. Microbial fermentation in the gastrointestinal tract, forestomach, or rumen produces volatile fatty acids (VFAs), such as acetic acid, propionic acid, and butyric acid. These fatty acids are absorbed directly through the gastrointestinal (GI) tract and / or rumen wall and used as feedstock, milk components, and other end-of-digestion products for the growth and development of the animal. The majority of the energy consumed by body tissues is used to produce milk, milk components, or muscle. Therefore, improved energy utilization can increase milk production, e.g., milk yield, and / or milk fat, milk protein, and / or milk solids. Improvements in body composition, such as increased muscle and / or changes in muscle / fat ratio in animals, can also be achieved.

[0151] Feed efficiency can be calculated by dividing the weight of milk produced by an animal, or the live weight of an animal by the weight of dry matter consumed by the animal. Thus, animals with higher feed efficiency will produce more milk, milk with a higher content of milk components such as, but not limited to, fat and protein, and / or exhibit increased weight gain compared to animals with lower feed efficiency when given the same nutrient inputs. Feed efficiency can be measured by the difference in the growth of an animal by any of the following parameters: average daily gain, total gain, feed conversion ratio including both feed:gain and gain:feed, feed efficiency, mortality rate, and feed intake. That is, improved feed efficiency means that the feed intake / muscle gain ratio is reduced. Improved feed efficiency also means that the muscle weight gain / feed intake ratio is increased. The term feed efficiency can also refer to feed intake / weight gain or weight gain / feed intake. Feed efficiency can be standardized to account for differences in protein and fat content by using energy corrected milk (ECM) yield instead of milk weight. This can be calculated using the following formula (Tyrrell and Reid, 1965): ECM = (12.82 x fat weight (lbs)) + (7.13 x protein weight (lbs)) + (0.323 x milk weight (lbs))

[0152] "Feed conversion" and "residual feed intake (RFI)" are also commonly used measures of feed efficiency, and the terms are often used interchangeably. In animal husbandry, feed conversion or feed conversion ratio is the ratio or rate measurement of efficiency with which an animal's body converts the animal's feed into a desired output. RFI is defined as the difference between an animal's actual dry matter intake (DMI) and the expected DMI required for maintenance and growth.

[0153] The primary benefit of improving feed efficiency (i.e., improving feed conversion ratio or lowering RFI) is to reduce the DMI of the animals without compromising growth performance, as feed-related costs are often the largest production expense in beef or milk production. Any reduction in DMI to produce one unit of beef or dairy results in minimizing feed costs and maximizing overall profitability for the beef or dairy industry.

[0154] In one embodiment, the feed efficiency in the ruminant animal is increased by at least about 1.01 times, such as at least about 1.02 times, 1.03 times, 1.04 times, 1.05 times, 1.06 times, 1.07 times, 1.08 times, 1.09 times, 1.10 times, 1.12 times, 1.14 times, 1.16 times, 1.18 times, such as at least about 1.20 times, of the feed efficiency of an untreated animal.

[0155] Increased feed efficiency may result from changes in the volatile fatty acid (VFA) profile, total VFA concentration and / or fermentation rate in the rumen and forestomach.

[0156] In some embodiments, L. rhamnosus FNZ129 or its derivatives promote propionic acid production. Propionic acid has a higher ATP production efficiency than other volatile fatty acids, which promotes propionic acid production and improves feed efficiency. Propionic acid is also glucogenic, and therefore can promote lactose synthesis in the mammary gland.

[0157] In some embodiments, L. rhamnosus FNZ129 or a derivative thereof shifts hydrogen metabolism from methane production to short chain / volatile fatty acid (VFA) production, such as propionic acid production. Propionic acid is primarily used as a precursor to glucose, and the formation of more propionic acid appears to result in more efficient utilization of feed energy. Maximizing the flow of metabolic hydrogen in the forestomach or rumen away from methane and toward VFAs (mainly propionic acid) increases the efficiency and reduces the environmental impact of livestock production (e.g., ruminant production), and enhances rumen development and / or ruminal papilla development.

[0158] Together with β-hydroxybutyrate, which is produced when butyrate is taken up, acetate is the major substrate for mammary lipogenesis. As a result, a high acetate fermentation pattern provides substrates to maintain or increase milk fat.

[0159] Thus, in some embodiments, the L. rhamnosus strain FNZ129 or a derivative thereof provides increased milk fat, milk protein, total milk yield and / or milk solids as a result of increased VFAs in the forestomach or rumen, which can act as an increased energy source to drive increased production.

[0160] In some embodiments, the yield of milk and / or milk components produced from the animal is preferably increased by at least 1.5%, more preferably at least 3.0%, at least 4.5%, or at least 6.0%.

[0161] In some embodiments, the L. rhamnosus strain FNZ129 or a derivative thereof results in increased live weight, muscle mass, and / or fat deposition, and / or improved body composition (e.g., a change in muscle / fat ratio) in the animal as a result of increased VFAs in the forestomach or rumen, which can act as an increased energy source to drive increased production.

[0162] In some embodiments, the live weight of the animal is preferably increased by at least 1%, more preferably at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, or at least 10% compared to a reference animal.

[0163] It is also expected that the present invention can be used to extend the lactation cycle of lactating ruminants, such as dairy cows. Dairy cows direct a significant portion of their energy toward producing milk during lactation. After a long lactation period, body condition deteriorates. For this reason, lactation periods are usually shortened or abbreviated to prevent excessive deterioration in body condition. It is expected that the methods and ruminant feed compositions disclosed herein will increase the feed efficiency of ruminants, thus reducing the impact of milk production on body condition. As a result, dairy cows can be milked for longer periods.

[0164] It is also expected that the present invention can be used to reduce or improve the deterioration of body condition due to lactation. It is expected that the methods and ruminant feed compositions disclosed herein will increase the feed efficiency by ruminants, thus resulting in ruminants with improved body condition at the end of lactation. For example, as an animal enters the dry period, the animal will have a higher body condition score (BCS). As a result, ruminants will require less dry matter intake in the off-season to achieve body condition. Alternatively or additionally, the methods and ruminant feed compositions disclosed herein are useful for improving the body condition of animals prior to lactation. For example, the methods and compositions disclosed herein can improve the body composition of the mother and / or fetus or newborn. For example, the methods and compositions disclosed herein can improve the body composition and / or weight of the newborn at birth.

[0165] It is also expected that the methods of the present invention can be used to reduce or improve the deterioration of body condition due to parturition or egg laying. It is expected that the methods disclosed herein will increase the feeding efficiency by monogastric animals, thus resulting in monogastric animals with improved body condition at the end of parturition or egg laying. As a result, monogastric animals will require less feed intake to achieve body condition. Alternatively or additionally, the methods and feed compositions disclosed herein are useful for improving the body condition of pre-lactation animals. For example, the methods and compositions disclosed herein can improve the body composition of the mother and / or fetus or newborn. For example, the methods and compositions disclosed herein can improve the body composition and / or weight of the newborn at birth.

[0166] It is also anticipated that the present invention may be similarly useful for reducing or ameliorating deterioration in body condition during other stresses such as parturition, drought, or inadequate feed intake.

[0167] As discussed above, the methods and compositions disclosed herein enhance the physical and / or functional development of the rumen, particularly in the early life of young or preweaned ruminants. Rumen development involves three distinct processes: (i) anatomical development (e.g., ruminal mass growth and ruminal papilla growth), (ii) functional achievement (e.g., fermentative capacity and enzyme activity), and (iii) microbial colonization (bacteria, fungi, methanogens, and protozoa).

[0168] Anatomical development of the rumen is a process that occurs following three stages: non-ruminant (0–3 weeks), transitional (3–8 weeks), and ruminal (from 8 weeks). During the transitional stage, growth and development of the ruminal absorptive surface area (papillae) is essential to allow absorption and utilization of digestive end products, especially ruminal volatile fatty acids. The presence and absorption of volatile fatty acids stimulates ruminal epithelial metabolism and may be key in the initiation of ruminal epithelial development. Constant exposure to volatile fatty acids maintains the development, size, and function of ruminal papillae. Different volatile fatty acids stimulate such growth differently, with butyric acid being the most stimulating, followed by propionic acid. Thus, a shift in hydrogen metabolism from methane production to short-chain / volatile fatty acid (VFA) production, e.g., propionic acid production, is expected to enhance ruminal epithelial growth and development.

[0169] As used herein, the term "gastrointestinal tract" refers to the portion of the digestive system beginning with the stomach and ending with the rectum, including the small intestine. Thus, for purposes of this application, the oral cavity and esophagus are not considered part of the digestive tract.

[0170] In some embodiments, the growth of methanogenic bacteria and / or archaea is inhibited in the feces of the animal. In some embodiments, the growth of methanogenic bacteria and / or archaea is inhibited in the distal gut of the animal. In some embodiments, the growth of methanogenic bacteria and / or archaea is inhibited in the colon of the animal. In some embodiments, the growth of methanogenic bacteria and / or archaea is inhibited in the rectum of the animal. In some embodiments, the growth of methanogenic bacteria and / or archaea is inhibited in the small intestine of the animal. In some embodiments, the growth of methanogenic bacteria and / or archaea is inhibited in the hindgut of the animal. In some embodiments, the growth of methanogenic bacteria and / or archaea is inhibited in the cecum of the animal.

[0171] It is also expected that the methods of the present invention may be useful for improving intestinal comfort or for preventing, reducing or ameliorating symptoms caused by gases produced by methanogens in the gastrointestinal tract of an animal, such as excessive flatulence, abdominal distension (bloating) and abdominal pain.

[0172] Ruminants In some embodiments, the food or feed composition is a ruminant feed composition. In some embodiments, the animal is a ruminant.

[0173] Ruminants are a group of herbivorous animals with a multi-compartment stomach that digests food by first microbial fermentation in the rumen to form the rumen, expels the rumen for chewing, and then swallows the chewed rumen for further digestion. This group includes, but is not limited to, the ruminants and the suborder Tylopoda, including several species of livestock. In one embodiment, the ruminant is a cow, goat, sheep, bison, yak, buffalo, deer, camel, alpaca, llama, wildebeest, antelope, or nilgai. In a preferred embodiment, the ruminant is a cow or a sheep.

[0174] In one embodiment, the ruminant is a lactating animal, hi an alternative embodiment, the ruminant is a pre-weaned animal, such as a calf or lamb.

[0175] The rumen is divided into the non-glandular forestomach (rumen, reticulum, and omasum) and the terminal glandular stomach, the abomasum.

[0176] In one embodiment, the ruminant is a newborn, neonatal, or juvenile. For example, in some embodiments, the ruminant is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, or 2 months old.

[0177] In some embodiments, the L. rhamnosus FNZ129 strain or derivative thereof is administered to the ruminant prior to weaning. In some embodiments, the L. rhamnosus FNZ129 or derivative thereof is administered to the ruminant after weaning. In some embodiments, the L. rhamnosus FNZ129 or derivative thereof is administered to the ruminant both prior to and after weaning. For example, in some embodiments, the L. rhamnosus FNZ129 strain or derivative thereof is administered throughout the life of the ruminant.

[0178] For example, L. rhamnosus FNZ129 or a derivative thereof is administered to a ruminant on or about postnatal day 0, e.g., on or about postnatal day 0, 1 or 2. Administration may then be given at least once a day, e.g., multiple times a day, sufficient to provide a sustained effect. For example, administration may continue for 2, 3, 4, 5, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 6 weeks, 2 months, 10 weeks, 3 months or more from birth. In some embodiments, administration of the L. rhamnosus strain FNZ129 or a derivative thereof continues for 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 years, 15 years, 16 years, 17 years, 18 years, 19 years, 20 years, or for the life of the animal.

[0179] monogastric animals In some embodiments, the food composition or feed composition is a feed composition for a monogastric animal. In some embodiments, the animal is a monogastric animal.

[0180] Monogastric animals are a group of animals that have a single chamber stomach, as compared to ruminants, which have a stomach containing multiple compartments, including the foregut or rumen. Monogastric animals include carnivores, omnivores, and herbivores, such as humans, cats, dogs, pigs, horses, donkeys, rabbits, and poultry.

[0181] Monogastric animals include some species of livestock. In one embodiment, the monogastric animal is a human, a pig, a horse, a donkey, a rabbit, or a poultry. In a preferred embodiment, the monogastric animal is a pig. In one embodiment, the monogastric animal is a chicken, a duck, a goose, or a turkey. In one embodiment, the monogastric animal is a companion animal, such as a cat or a dog.

[0182] In one embodiment, the monogastric animal is a pre-weaned animal, such as a piglet or a foal. In some embodiments, L. rhamnosus FNZ129 or a derivative thereof is administered to the monogastric animal before weaning. In some embodiments, L. rhamnosus FNZ129 or a derivative thereof is administered to the monogastric animal after weaning. In some embodiments, L. rhamnosus FNZ129 or a derivative thereof is administered to the monogastric animal both before and after weaning. For example, in some embodiments, the L. rhamnosus FNZ129 strain or a derivative thereof is administered throughout the life of the animal.

[0183] For example, L. rhamnosus FNZ129 or a derivative thereof is administered to the animal on or about birth day 0, e.g., on or about birth day 0, 1 or 2. Administration may then be given at least once a day, e.g., multiple times a day, sufficient to provide a sustained effect. For example, administration may continue for 2, 3, 4, 5, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 6 weeks, 2 months, 10 weeks, 3 months or more from birth. In some embodiments, administration of the L. rhamnosus strain FNZ129 or a derivative thereof continues for 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 years, 15 years, 16 years, 17 years, 18 years, 19 years, 20 years, or for the life of the animal.

[0184] Lacticaseibacillus rhamnosus FNZ129 A culture of Lacticaseibacillus rhamnosus FNZ129 (also known as Lactobacillus rhamnosus FNZ129) was isolated from a human source and deposited on August 2, 2021 at the National Measurement Institute of Australia (NMIA), 1 / 153 Bertie Street, Port Melbourne, Victoria, Australia 3207, and was accorded the accession number V21 / 015446, an international depository recognized under the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. The terms Lactobacillus rhamnosus strain FNZ129, Lactobacillus rhamnosus FNZ129, Lacticaseibacillus rhamnosus FNZ129, and L. rhamnosus FNZ129 are used interchangeably herein.

[0185] Whole-genome sequencing using a combination of short-read (Illumina) and long-read (PacBio) sequencing technologies was used to generate a hybrid genome assembly. The final hybrid assembly contained two contigs. The total length was 3,094,881 bp (3.09 Mb). The species ID for strain FNZ129 was confirmed as Lacticaseibacillus rhamnosus using the taxonomic sequence classification program Kraken.

[0186] All working groups and associated bioinformatics were carried out in accordance with the EFSA guidelines available at https: / / efsa.onlinelibrary.wiley.com / doi / pdf / 10.2903 / j.efsa.2018.5206 and the latest EFSA statement (July 2021) available at https: / / efsa.onlinelibrary.wiley.com / doi / full / 10.2903 / j.efsa.2021.6506

[0187] Morphological properties The morphological characteristics of L. rhamnosus FNZ129 are described below.

[0188] When grown in MRS broth, they are short to medium rod-shaped with square ends in chains, typically 0.7 x 1.1 x 2.0-4.0 μm in size. They are gram-positive, non-mobile, non-spore-forming, catalase-negative facultative anaerobic rods.

[0189] Further characterization Of course, there are methods widely known and available to those of skill in the art that can be used to confirm the identity of L. rhamnosus FNZ129; exemplary methods include DNA fingerprinting, genomic analysis, sequencing, and related genomic and proteomic techniques.

[0190] L. rhamnosus strain FNZ129 and its derivatives As described herein, certain embodiments of the invention utilize live L. rhamnosus strain FNZ129. In other embodiments, derivatives of L. rhamnosus strain FNZ129 are utilized.

[0191] As used herein, the term "derivatives" and its grammatical equivalents when used in reference to bacteria (including when used in reference to specific strains of bacteria such as L. rhamnosus FNZ129) contemplates mutants and homologs derived from bacteria, killed or attenuated bacteria, such as, but not limited to, heat-killed, lysed, fractionated, pressure-killed, irradiated, and UV- or light-treated bacteria, as well as antimethanogenic factors from bacteria, bacterial metabolites, bacterial cell suspensions, bacterial culture supernatants, and the like, where the derivatives retain antimethanogenic activity. Transgenic microorganisms engineered to express one or more antimethanogenic factors are also contemplated. Methods for producing such derivatives, such as, but not limited to, one or more mutants or one or more antimethanogenic factors of L. rhamnosus strain FNZ129, particularly derivatives suitable for administration to ruminants (e.g., in compositions), are well known in the art.

[0192] It will be appreciated that methods suitable for identifying L. rhamnosus strain FNZ129 are equally suitable for identifying mutants or homologs of L. rhamnosus strain FNZ129, or derivatives of L. rhamnosus strain FNZ129, for example, including bacterial metabolic products from L. rhamnosus strain FNZ129.

[0193] The term "antimethanogenic factors" refers to bacterial molecules responsible for mediating antimethanogenic activity, including but not limited to bacterial DNA motifs, RNA (including mRNA and miRNA), proteins, exosomes, bacteriocins, bacteriocin-like molecules, antimicrobial peptides, antibiotics, antimicrobial agents, small molecules, polysaccharides, or cell wall components such as lipoteichoic acid and peptidoglycan, or mixtures of any two or more thereof. As noted above, these molecules have not been definitively identified and without wishing to be bound by any theory, their presence can be inferred by the presence of antimethanogenic activity.

[0194] The term "antimethanogenic activity" refers to the ability of a particular microorganism to inhibit the growth of methanogenic bacteria and / or archaea and / or to reduce the production of methane by methanogenic bacteria and / or archaea. This ability can be limited to inhibiting the growth and / or methane producing capacity of a particular group of methanogenic bacteria and / or archaea, for example inhibiting the growth of hydrogenotrophic methanogens, inhibiting the methane producing capacity of hydrogenotrophic methanogens, inhibiting the growth of methylotrophic methanogens, inhibiting the methane producing capacity of methylotrophic methanogens, inhibiting the growth of a particular species of methanogens, or inhibiting the methane producing capacity of a particular species of methanogens.

[0195] Reference to retaining anti-methanogen activity is intended to mean that a derivative of the microorganism, such as a mutant or homologue of the microorganism, or an attenuated or killed microorganism, or a cell culture supernatant, still has useful anti-methanogen activity, or that a composition comprising the microorganism or a derivative thereof still has useful anti-methanogen activity. Although the bacterial molecules responsible for mediating the anti-methanogen activity have not been definitively identified, molecules that have been proposed as possible candidates include bacterial DNA motifs, RNA including mRNA and miRNA, proteins, exosomes, bacteriocins, antibiotics, surface proteins, small organic acids, polysaccharides, and cell wall components such as lipoteichoic acid and peptidoglycan. These are hypothesized to interact with components of methanogenic bacteria and / or archaea to provide a growth inhibitory effect. Preferably, the retained activity is at least about 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99 or 100% of the activity of an untreated (i.e., live or non-attenuated) control, and useful ranges can be selected between any of these values ​​(e.g., about 35 to about 100%, about 50 to about 100%, about 60 to about 100%, about 70 to about 100%, about 80 to about 100%, and about 90 to about 100%).

[0196] L. rhamnosus strain FNZ129 can be grown in sufficient quantities to permit use as contemplated herein using conventional solid substrate and liquid fermentation techniques well known in the art. For example, L. rhamnosus FNZ129 can be grown using nutrient film or submerged culture growth techniques, e.g., as described in WO 99 / 10 4 76, can be produced in bulk for formulation. Briefly, growth is performed under aerobic conditions at any temperature sufficient for the growth of the organism. For example, for L. rhamnosus FNZ129 strain, a temperature range of 30-40°C, preferably 37°C, is preferred. The pH of the growth medium is slightly acidic, preferably about 6.0-6.5. The incubation time is sufficient for the isolate to reach stationary growth phase.

[0197] The bacterial cells may be harvested by methods well known in the art, such as conventional filtration or sedimentation methods (e.g., centrifugation), or harvested in a dry state using a cyclone system. The bacterial cells may be used immediately or stored for as long as required using standard techniques, preferably freeze-dried or chilled at -20°C to 6°C, preferably -4°C. Cryoprotectants, cryopreservatives, and / or cryoprotectants may also be used to increase the stability and / or viability of the bacterial cells when dried and / or frozen, as known in the art.

[0198] Supernatant Further embodiments of the present invention utilize supernatants from cell cultures comprising L. rhamnosus strain FNZ129 or derivatives thereof. These embodiments include processes for preparing bacterial culture supernatants, said processes comprising culturing bacterial cells and separating the supernatant from the cultured cells, thereby obtaining a supernatant. This method also allows for further isolation of bacterial molecules responsible for mediating anti-methanogen activity, which can be obtained from the supernatant.

[0199] As will be appreciated by one of skill in the art, supernatants useful in the present invention include both supernatants from such cultures, and / or concentrates of such supernatants and / or fractions of such supernatants.

[0200] The term "supernatant" in this context refers to the medium from a bacterial culture from which the bacteria have subsequently been removed, for example by centrifugation or filtration.

[0201] The supernatants useful in the present invention can be readily obtained by a simple process for preparing bacterial culture supernatants, which process comprises: a) Cultivation of cells of L. rhamnosus strain FNZ129; b) optionally, the release of active compounds and / or extracellular components of the cells by various cell treatments, such as, but not limited to, acidic or alkaline modification, sonication, detergents, e.g. sodium dodecyl sulfate (SDS) and / or Triton X, wall-soluble enzymes such as mutarolysin and / or lysozyme, salts and / or alcohols, and c) Isolation of supernatant from cultured cells thereby obtaining said supernatant liquid.

[0202] In a preferred embodiment of this process, the supernatant composition is further subjected to a drying step to obtain a dried culture product.

[0203] The drying step may conveniently be freeze-drying or spray-drying, but any drying process suitable for drying antimethanogenic agents such as bacteriocins is contemplated, including vacuum drying and air drying.

[0204] Although the contents of the supernatant produced by L. rhamnosus strain FNZ129 have not yet been characterized in detail, it is known that certain bacterial strains can produce bacteriocins, which are small thermostable proteins, and therefore, without wishing to be bound by theory, it is expected that drying methods, including spray drying, which result in moderate heating of the culture eluate product, will also result in active compositions, as demonstrated in the examples described herein.

[0205] lysate The liquid containing the contents of lysed cells is called a lysate. The lysate contains the active components of the bacterial cells and can be either crude, i.e. containing all cellular components, or partially and / or completely separated into separate fractions such as extracellular components, intracellular components, proteins, etc.

[0206] Methods for producing bacterial cell lysates are well known in the art. Such methods may include, but are not limited to, mechanical lysis, such as mechanical shearing, grinding, milling, or sonication, enzymatic lysis, such as enzymes that degrade bacterial cell walls, chemical lysis, such as using detergents, denaturants, pressure changes, and / or osmotic shock, and combinations of the above.

[0207] Thus, a further embodiment of the present invention utilises a lysate of L. rhamnosus strain FNZ129 or a derivative thereof.

[0208] Cell suspension The present invention may also, in some embodiments, utilize a cell suspension comprising L. rhamnosus strain FNZ129 or a derivative thereof.

[0209] In the present context, the term "cell suspension" relates to a plurality of cells of L. rhamnosus strain FNZ129 or a derivative thereof dispersed or suspended in a liquid, such as a liquid nutrient medium, culture medium or saline solution.

[0210] The cells may be provided in the form of a cell suspension in a suitable solution for dispersion, which may be dispersed, for example, by spraying, dipping, or any other application process.

[0211] The cells may be viable, but the suspension may also contain inactivated or dead cells or their lysates. In one embodiment, the suspension of the present invention contains live cells. In another embodiment, the suspension of the present invention contains inactivated, dead or lysed cells.

[0212] Bacteriocins Bacteriocins are antibacterial compounds produced by bacteria to inhibit other bacterial strains and species.

[0213] Lactic acid bacteria (LAB) are well known to produce bacteriocins, and these compounds are of global interest to the food industry because they inhibit the growth of many spoilage and pathogenic bacteria, thus extending the shelf life and safety of foods. Bacteriocins are generally considered to be narrow-spectrum antibiotics. Moreover, bacteriocins, especially from LAB, show very low human toxicity and have been consumed in fermented foods for thousands of years.

[0214] A further aspect of the invention provides an isolated antibacterial compound obtained from L. rhamnosus strain FNZ129 or a derivative thereof. Such an antibacterial compound may, for example, be obtained from a supernatant or lysate resulting from a process described herein further comprising an isolation step.

[0215] As illustrated in the Examples disclosed herein, L. rhamnosus strain FNZ129 and / or compositions comprising L. rhamnosus strain FNZ129 and / or culture supernatants of L. rhamnosus strain FNZ129 have been found to be useful as antimicrobial compounds, particularly for inhibiting the growth of methanogenic bacteria and / or for inhibiting the ability of methanogens to produce methane.

[0216] In the present context, the term antimicrobial compound utilizes compounds that kill, impair the survival or inhibit the growth of microorganisms.

[0217] Antibacterial compounds can be classified according to the microorganisms they primarily act on: for example, antibacterial agents are used against bacteria and antifungals against fungi. They can also be classified according to their function: compounds that kill microorganisms are called bacteriocidal, those that only inhibit their growth are called bacteriostatic.

[0218] In one embodiment, the invention relates to antimicrobial compounds that are bactericidal. In another embodiment, the invention relates to antimicrobial compounds that are bacteriostatic. In another embodiment, the invention relates to antimicrobial compounds that are antimicrobial.

[0219] Feed or carrier composition Feed compositions useful herein (e.g., ruminant feed compositions or monogastric feed compositions) can be formulated as a food, beverage, food additive, beverage additive, animal feed, animal feed additive, animal feed supplement, dietary supplement, carrier, vitamin or mineral premix, nutritional product, enteral feed product, soluble, slurry, supplement, pharmaceutical, lick block, drench, tablet, capsule, pellet or intraruminal product, e.g., bolus. Suitable formulations can be prepared by one of ordinary skill in the art having regard to the technology and the teachings of this specification.

[0220] The composition can be administered as a top dressing or mixed into standard feed ingredients such as the daily ration. Additionally, the strain can be administered in partial or total mixed feeds (TMR), pelleted feeds, mixed with liquid feeds or beverages, mixed with protein premixes, or delivered via vitamin and mineral premixes.

[0221] In one embodiment, the compositions useful herein include any edible feed product capable of carrying bacteria or bacterial derivatives. As used in this application, the term "feed" or "animal feed" refers to a substance consumed by an animal that contributes energy and / or nutrients to the animal's diet. Animal feed typically includes many different ingredients that may be present in the form of concentrates, premixes, co-products, or pellets. Examples of feeds and feed ingredients include partial or total mixed rations (TMR), corn, soybeans, forage, grains, distillers grains, germinated grains, legumes, vitamins, amino acids, minerals, fiber, forage, grass, hay, straw, silage, grains, leaves, meals, solubles, slurries, supplements, mash meals, meals, fruit pulps, vegetable pulps, fruit or vegetable pomace, citrus meal, wheat shorts, corn cob meal, molasses, and the like. Other compositions useful as carriers include milk, milk powder, milk replacers, milk fortifiers, colostrum, whey, whey powder, sucrose, maltodextrin, rice hulls, and the like.

[0222] In certain embodiments, the feed composition is formed through a process of growing L. rhamnosus strain FNZ129 using a milk-based carrier, such as thermalized milk, or a non-milk-based carrier, to create a fermented yogurt-style composition. Methods of creating such fermented yogurt-style compositions are well known in the art and may include, for example, incubating the milk at an appropriate temperature, using a hot water bath or other heating means, for example, for 12 hours or more until sufficient cell density is reached. In one embodiment, the temperature is 25-30°C. Optionally, the milk may contain other additives to promote bacterial growth, such as yeast extract. In certain embodiments, the method is performed in the field, such as a farm, where probiotic feed supplementation is performed. The fermented yogurt-style composition may be administered by oral application, such as drenching. In some embodiments, the fermented yogurt-style composition is administered at a dose of 1-100 ml per day, for example, 2-50, 5-30, or 10-20 ml per day.

[0223] Other suitable feed formulations for ruminants are described in E. W. Crampton et al., Applied Animal Nutrition, WH Freeman and Company, San Francisco, Calif., 1969 and DC Church, Livestock Feeds and Feeding, 0 & B Books, Corvallis, Oreg., 1977, both of which are incorporated herein by reference.

[0224] In one embodiment, compositions useful herein include any non-feed carrier that is consumed by the animal to which the bacteria or bacterial derivatives are added, such as vermiculite, zeolite, or crushed limestone.

[0225] In one embodiment, the compositions useful herein include pet food compositions for companion animals such as cats and dogs. In a particular embodiment, L. rhamnosus FNZ129 is present at about 10 4 cfu (colony forming units) / g of pet food ~ approx. 10 14 cfu / g of the pet food. In certain embodiments, the composition further comprises at least one protein source. In certain embodiments, the composition further comprises at least one fat source. In certain embodiments, the composition further comprises at least one carbohydrate source. In certain embodiments, the pet food is a dog food. In certain embodiments, the pet food is a cat food.

[0226] The term "pet food" or "pet food composition" as used herein means a nutritional composition intended for consumption by a pet. In one embodiment, a nutritional composition may refer to a dietary supplement intended for consumption by a pet. A dietary supplement is intended to refer to a composition that provides nutrients that are not otherwise consumed in sufficient amounts by a pet. In one embodiment, a nutritional composition may refer to a pet treat intended for consumption by a pet. The term "pet treat" as used herein refers to food for consumption by a pet that is intended as an occasional reward or indulgence, rather than as the sole source of nutrition for the pet.

[0227] In one embodiment, the compositions useful herein include food compositions for omnivores such as chickens, pigs, humans, and dogs. Such food compositions are well known in the art.

[0228] In a particular embodiment, the compositions of the invention comprise live L. rhamnosus strain FNZ 129. Methods for producing such compositions are well known in the art.

[0229] In some embodiments, the compositions of the invention comprise one or more derivatives of the L. rhamnosus FNZ129 strain. Again, methods for producing such compositions are well known in the art and can utilize standard microbiological and pharmaceutical practices. In some embodiments, the compositions comprise a dried culture product, such as a supernatant or a cell lysate, as described herein.

[0230] It is understood that a wide range of additives or carriers can be included in such compositions, for example, to improve or preserve bacterial viability or to increase the anti-methanogen activity of L. rhamnosus strain FNZ129 or its derivatives. For example, additives such as surfactants, wetting agents, humectants, stickers, dispersants, stabilizers, penetrants, and so-called stress additives (such as potassium chloride, glycerol, sodium chloride, and glucose) to improve the vitality, growth, replication, and survival of bacterial cells, as well as cryoprotectants such as maltodextrin, may be included. Additives may also include compositions that help maintain the viability of microorganisms in long-term storage, such as unrefined corn oil, or "inverse" emulsions that include a mixture of oil and wax on the outside and water, sodium alginate, and bacteria on the inside.

[0231] In some embodiments, L. rhamnosus FNZ129 or a derivative thereof is encapsulated. Methods for producing such encapsulated bacteria are well known in the art. In some embodiments, L. rhamnosus FNZ129 or a derivative thereof is encapsulated in liposomes, microbubbles, microparticles, microcapsules, or the like. Such encapsulating agents include natural, semi-synthetic, or synthetic polymers, waxes, lipids, fats, fatty alcohols, fatty acids, and / or plasticizers, such as alginates, gums, kappa-carrageenan, chitosan, starch, sugar, gelatin, and the like.

[0232] In a particular embodiment, the L. rhamnosus strain FNZ129 is in a reproductively viable form and amount.

[0233] The composition may include a carbohydrate source, such as disaccharides, including, for example, sucrose, fructose, glucose, or dextrose. Preferably, the carbohydrate source is one that can be utilized by the L. rhamnosus FNZ129 strain aerobically or anaerobically.

[0234] In such embodiments, the composition is capable of supporting the reproductive viability of L. rhamnosus strain FNZ129 for a period of greater than about 2 weeks, preferably greater than about 1 month, greater than about 2 months, greater than about 3 months, greater than about 4 months, greater than about 5 months, more preferably greater than about 6 months, and most preferably for at least about 2 years to about 3 years or more.

[0235] In certain embodiments, the oral composition is formulated to administer an effective amount of L. rhamnosus strain FNZ129 to allow for establishment of a population in the gastrointestinal tract of an animal upon ingestion. The established population may be a temporary or permanent population.

[0236] While a variety of routes and methods of administration are contemplated, oral administration of L. rhamnosus strain FNZ129, e.g., in a composition suitable for oral administration, is currently preferred, although it will of course be recognized that other routes and methods of administration may be utilized or may be preferred in particular circumstances.

[0237] The term "oral administration" includes oral, buccal, enteral, intraruminal, and intragastric administration.

[0238] Theoretically, one colony forming unit (cfu) should be sufficient to establish a population of L rhamnosus strain FNZ129 in an animal, but in practical situations, a minimum number of units is required to do so. Thus, for therapeutic mechanisms that rely on a viable live population of probiotic bacteria, the number of units administered to a subject will affect efficacy.

[0239] In one embodiment, the formulation formulated for administration contains at least about 6×109 cfu / day, e.g., at least about 6×10 11 In another embodiment, the formulation formulated for administration is sufficient to provide at least about 10 cfu / day of L. rhamnosus FNZ129 strain. 10 In another embodiment, the formulation formulated for administration is sufficient to provide at least about 10 cfu of L. rhamnosus FNZ129 daily. 12 This is sufficient to provide cfu of L. rhamnosus FNZ129 strain.

[0240] Methods for determining the presence of a population of intestinal and / or ruminal flora (e.g., L. rhamnosus strain FNZ129) in the gastrointestinal tract of a subject are well known in the art, and examples of such methods are provided herein. In certain embodiments, the presence of a population of L. rhamnosus strain FNZ129 can be determined directly, for example, by analyzing one or more samples obtained from the animal and determining the presence or amount of L. rhamnosus strain FNZ129 in said samples. In other embodiments, the presence of a population of L. rhamnosus strain FNZ129 can be determined indirectly, for example, by observing a decrease in methane emission or methane production, a decrease in hydrogen production, or a decrease in the number of other intestinal and / or ruminal flora in a sample obtained from the animal. Combinations of such methods are also contemplated.

[0241] The efficacy of the composition useful according to the present invention can be evaluated both in vitro and in vivo. For example, see the following examples. Briefly, the composition can be tested for its ability to inhibit the growth of methanogenic bacteria and / or archaea, or its ability to reduce the production of methane by methanogenic bacteria and / or archaea. In in vivo studies, the composition can be fed or injected into ruminants or monogastric animals, and its effect on methanogenic bacteria and / or archaea, as well as its effect on methane emission, can be evaluated. Based on the results, the appropriate dosage range and administration route can be determined.

[0242] The method of calculating the appropriate dose may depend on the nature of the active agent in the composition.For example, if the composition contains live bacteria, the dose may be calculated by referring to the number of live bacteria present.For example, as described in the examples herein, the dose may be established by referring to the number of colony forming units (cfu) to be administered per day, or by referring to the number of cfu per kilogram of dry feed weight.

[0243] For example, approximately 1 × 10 of L. rhamnosus strain FNZ129 per kg of dry feed weight. 6 cfu ~ approx. 1 × 10 12 cfu / day, preferably about 1×10 6 cfu ~ approx. 1 × 10 11 cfu / kg / day, approximately 1 × 10 6 cfu ~ approx. 1 × 10 10 cfu / kg / day, approximately 1 × 10 6 cfu ~ approx. 1 × 10 9 cfu / kg / day, approximately 1 × 10 6 cfu ~ approx. 1 × 10 8 cfu / kg / day, approximately 1 × 10 6 cfu ~ approx. 5 × 10 7 cfu / kg / day, or approximately 1 x 10 6 cfu ~ approx. 1 × 10 7 A dosage of about 5×10 cfu / kg / day of L. rhamnosus strain FNZ129 per kg of dry material weight is contemplated. 6 cfu ~ approx. 5 × 10 8 cfu / day, preferably about 5×10 6 cfu ~ approx. 4 × 10 8 cfu / kg / day, approximately 5 × 10 6 cfu ~ approx. 3 × 10 8 cfu / kg / day, approximately 5 × 10 6 cfu ~ approx. 2 × 10 8 cfu / kg / day, approximately 5 × 10 6 cfu ~ approx. 1 × 10 8 cfu / kg / day, approximately 5 × 10 6 cfu ~ approx. 9 × 10 7 cfu / kg / day, approximately 5 × 10 6 cfu ~ approx. 8 × 10 7cfu / kg / day, approximately 5 × 10 6 cfu ~ approx. 7 × 10 7 cfu / kg / day, approximately 5 × 10 6 cfu ~ approx. 6 × 10 7 cfu / kg / day, approximately 5 × 10 6 cfu ~ approx. 5 × 10 7 cfu / kg / day, approximately 5 × 10 6 cfu ~ approx. 4 × 10 7 cfu / kg / day, approximately 5 × 10 6 cfu ~ approx. 3 × 10 7 cfu / kg / day, approximately 5 × 10 6 cfu ~ approx. 2 × 10 7 cfu / kg / day, or approximately 5 × 10 6 cfu ~ approx. 1 × 10 7 Dosing at cfu / kg / day is contemplated.

[0244] In certain embodiments, the periodic dose need not vary with the subject's body weight, dry food weight, or other characteristics. In such examples, about 1×10 6 cfu ~ approx. 1 × 10 13 cfu / day, preferably about 1×10 6 cfu ~ approx. 1 × 10 12 cfu / day, approximately 1 × 10 6 cfu ~ approx. 1 × 10 11 cfu / day, approximately 1 × 10 6 cfu ~ approx. 1 × 10 10 cfu / day, approximately 1 × 10 6 cfu ~ approx. 1 × 10 9 cfu / day, approximately 1 × 10 6 cfu ~ approx. 1 × 10 8 cfu / day, approximately 1 × 10 6 cfu ~ approx. 5 × 10 7 cfu / day, or approximately 1 × 10 6 cfu ~ approx. 1 × 10 7 Dosing of cfu / day is contemplated.

[0245] In one embodiment, about 5×10 L. rhamnosus strain FNZ129 per kg body weight 7 cfu ~ approx. 5 × 10 10 cfu / day, preferably about 5×107 cfu ~ approx. 4 × 10 10 cfu / day, approximately 5 × 10 7 cfu ~ approx. 3 × 10 10 cfu / day, approximately 5 × 10 7 cfu ~ approx. 2 × 10 10 cfu / day, approximately 5 × 10 7 cfu ~ approx. 1 × 10 10 cfu / day, approximately 5 × 10 7 cfu ~ approx. 9 × 10 9 cfu / day, approximately 5 × 10 7 cfu ~ approx. 8 × 10 9 cfu / day, approximately 5 × 10 7 cfu ~ approx. 7 × 10 9 cfu / day, approximately 5 × 10 7 cfu ~ approx. 6 × 10 9 cfu / day, approximately 5 × 10 7 cfu ~ approx. 5 × 10 9 cfu / day, approximately 5 × 10 7 cfu ~ approx. 4 × 10 9 cfu / day, approximately 5 × 10 7 cfu ~ approx. 3 × 10 9 cfu / day, approximately 5 × 10 7 cfu ~ approx. 2 × 10 9 cfu / day, or approximately 5 × 10 7 cfu ~ approx. 1 × 10 9 cfu / day is contemplated. Preferably, 1×10 8 ~1×10 9 The dose is administered at cfu / kg body weight / day.

[0246] It is understood that in certain embodiments, the dosage does not need to be administered every day.For example, the composition can be formulated to be administered every 2 days, twice a week, once a week, every 2 weeks, or once a month.Alternatively, in certain embodiments, the composition can be formulated to be administered 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 times per day, with each meal, or with each mouthful.

[0247] In one embodiment, L. rhamnosus FNZ129 is present at about 1×10 per gram of pet food and / or pet food mixture.3 ~Approx. 1×10 9 It can be administered at cfu.

[0248] Suitably, L. rhamnosus FNZ129 is present at a concentration of about 1 x 10 per gram of pet food and / or pet food mixture. 4 ~Approx. 1×10 8 It can be administered at cfu.

[0249] Suitably, L. rhamnosus FNZ129 is present at a concentration of about 7.5 x 10 per gram of pet food and / or pet food mixture. 4 ~Approx. 1×10 7 It can be administered at cfu.

[0250] Preferably, L. rhamnosus FNZ129 is present at about 1 x 10 per gram of pet food and / or pet food mixture. 6 It can be administered at cfu.

[0251] In embodiments where the pet food is a pet treat, the number of cfu / g administered may be between about 2 and about 20 times the number of cfu / g administered in the pet food and / or pet food mix, suitably between about 4 and about 15 times the number of cfu / g administered in the pet food and / or pet food mix. Preferably, the number of cfu / g administered is about 10 times the number of cfu / g administered in the pet food and / or pet food mix.

[0252] It is understood that the composition is preferably formulated to allow administration of an effective dose of L. rhamnosus FNZ129 strain and / or one or more derivatives thereof. The dose of the composition administered, the duration of administration, and the general administration regimen may vary between animals, depending on the mode of administration selected, as well as variables such as the age, sex, weight, and species of the animal. Furthermore, as mentioned above, the appropriate dose may depend on the nature of the active agent in the composition and the mode of formulation.

[0253] In some embodiments, the dose of the composition does not change over time. In other embodiments, the dose of the composition may change over time. For example, in some embodiments, the initial dosing regimen may be followed by a maintenance dosing regimen. It is understood that a higher dose is required to establish a population of L. rhamnosus FNZ129 in an animal, and a lower dose is sufficient to maintain said population. Thus, in some embodiments, the initial dosing regimen includes administering a higher dose and / or a more frequent dose than the maintenance dosing regimen. The initial dosing schedule is preferably effective to establish a population of L. rhamnosus FNZ129 in an animal, and the maintenance dosing schedule is desirably effective to maintain a population of L. rhamnosus FNZ129 in an animal. In some embodiments, the maintenance dosing regimen includes administering a dose daily, every other day, twice weekly, twice weekly, every other week, or once monthly.

[0254] In some embodiments, the effects of the methods described herein persist after administration of L. rhamnosus FNZ129. Without wishing to be bound by theory, it is expected that administration of L. rhamnosus FNZ129 as described herein may result in long-lasting or even permanent changes in the forestomach or rumen of a ruminant animal, or the gastrointestinal tract of a monogastric animal. In some embodiments, the effects persist for 2 days after the last administration of L. rhamnosus FNZ129, for example, 3 days, 5 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, or 7 years after the last administration of L. rhamnosus FNZ129. In preferred embodiments, the effects persist for the life of the animal.

[0255] In the example where the composition comprises one or more derivatives of L. rhamnosus strain FNZ129, the dose can be calculated by reference to the amount or concentration of the derivative administered daily. For example, if the bacteria is to be inactivated, the aforementioned amount is calculated before inactivation. For a composition comprising L. rhamnosus strain FNZ129 culture supernatant, the dose can be calculated by reference to the concentration of the culture supernatant present in the composition. The concentration of the culture supernatant present in the composition can be calculated, for example, based on the cfu of the culture. For example, 1×10 9 The amount of culture supernatant equivalent to cfu / day can be calculated from the total yield of the culture and the total amount of culture supernatant.

[0256] It will be understood that the preferred compositions are formulated to provide an effective amount in a convenient form and amount. In certain embodiments, but not limited to, where the periodic dose does not need to vary with the weight or other characteristics of the animal, the composition can be formulated for a unit dose. Of course, administration includes a single daily administration, or multiple separate divided administrations as appropriate. For example, an effective dose of the L. rhamnosus FNZ129 strain can be incorporated into the feed for oral administration.

[0257] However, as a general example, we contemplate administration of from about 1 mg to about 1000 mg per day of the compositions useful herein, preferably from about 50 to about 500 mg per day, alternatively from about 150 to about 410 mg per day, or from about 110 to about 310 mg per day. In one embodiment, we contemplate administration of from about 0.05 mg to about 250 mg per kg of body weight of the compositions useful herein. For example, administration to humans may be as low as 6×10 for adults or children as a 500 mg capsule. 9 It may contain a single dose of CFU / day.

[0258] In one embodiment, the compositions useful herein comprise, consist essentially of, or consist of at least about 0.1, 0.2, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99, 99.5, 99.8, or 99.9% by weight of L. rhamnosus strain FNZ129 and / or derivatives thereof, and useful ranges range between any of these aforesaid values ​​(e.g., about 0.1 to about 50%, about 0.2 to about 50%, about 0.5 to about 50%). %, about 1 to about 50%, about 5 to about 50%, about 10 to about 50%, about 15 to about 50%, about 20 to about 50%, about 25 to about 50%, about 30 to about 50%, about 35 to about 50%, about 40 to about 50%, about 45 to about 50%, about 0.1 to about 60%, about 0.2 to about 60%, about 0.5 to about 60%, about 1 to about 60%, about 5 to about 60%, about 10 to about 60%, about 15 to about 60%, about 20 to about 60%, about 25 to about 60%, about 30 to about 60%, about 35 to about 60%, about 40 to about 60%, about 45 to about 60%, about 0.1 to about 70%, about 0.2 to about 70%, about 0 .5 to about 70%, about 1 to about 70%, about 5 to about 70%, about 10 to about 70%, about 15 to about 70%, about 20 to about 70%, about 25 to about 70%, about 30 to about 70%, about 35 to about 70%, about 40 to about 70%, about 45 to about 70%, about 0.1 to about 80%, about 0.2 to about 80%, about 0.5 to about 80%, about 1 to about 80%, about 5 to about 80%, about 10 to about 80%, about 15 to about 80%, about 20 to about 80%, about 25 to about 80%, about 30 to about 80%, about 35 to about 80%, about 40 to about 80%, about 45 to about 80%, about 0.1 to about 90%, about 0.2 to about 90%, about 0.5 to about 90%, about 1 to about 90%, about 5 to about 90%, about 10 to about 90%, about 15 to about 90%, about 20 to about 90%, about 25 to about 90%, about 30 to about 90%, about 35 to about 90%, about 40 to about 90%, about 45 to about 90%, about 0.1 to about 99%, about 0.2 to about 99%, about 0.5 to about 99%, about 1 to about 99%, about 5 to about 99%, about 10 to about 99%, about 15 to about 99%, about 20 to about 99%, about 25 to about 99%, about 30 to about 99%, about 35 to about 99%, about 40 to about 99%, and about 45 to about 99%).

[0259] In one embodiment, the compositions useful herein comprise, consist essentially of, or are at least about 0.001, 0.01, 0.05, 0.1, 0.15, 0.2, 0.3, 0.4, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 grams of L. rhamnosus strain FNZ129 and / or a derivative thereof. or consisting of, and useful ranges can be selected between any of these aforementioned values ​​(e.g., about 0.01 to about 1 gram, about 0.01 to about 10 grams, about 0.01 to about 19 grams, about 0.1 to about 1 gram, about 0.1 to about 10 grams, about 0.1 to about 19 grams, about 1 to about 5 grams, about 1 to about 10 grams, about 1 to about 19 grams, about 5 to about 10 grams, about 5 to about 19 grams).

[0260] In certain embodiments, the compositions useful herein have at least about 10 per kg dry weight of the composition. 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , 10 12 , or 10 13 The useful ranges include, consist essentially of, or consist of colony forming units (cfu), and may range from any of the values ​​above (e.g., about 10 5 ~about 10 13 cfu, approximately 10 6 ~about 10 12 cfu, approximately 10 7 ~about 10 12 cfu, approximately 10 8 ~about 10 11 cfu, approximately 10 8 ~about 10cfu, about 10 9 ~about 10 11 cfu).

[0261] It will be appreciated that the concentration of L. rhamnosus strain FNZ129 and / or one or more derivatives thereof in a composition formulated for administration may be lower than the concentration in a composition formulated for, for example, distribution or storage, and that the concentration of a composition formulated for storage and subsequently formulated into a composition suitable for administration must be sufficient to allow said composition for administration to be sufficiently concentrated so that it can be administered in an effective dose.

[0262] The compositions useful herein can be used alone or in combination with one or more other therapeutic agents. The therapeutic agent can be a food, beverage, food additive, beverage additive, food ingredient, beverage ingredient, dietary supplement, vitamin or mineral premix, oil, oil blend, oil-rich feed supplement, nutritional product, medical food, dietary supplement, medicine or pharmaceutical product. The therapeutic agent can be a probiotic agent or factor, preferably effective in inhibiting the growth of methanogenic bacteria and / or archaea or reducing methane release by methanogenic bacteria and / or archaea. In some embodiments, the oil, oil blend, or oil-rich feed supplement is palm kernel expeller (PKE) and / or PROLIQ.

[0263] When used in combination with another therapeutic agent, the administration of the composition useful herein and the other therapeutic agent can be simultaneous or sequential. Simultaneous administration includes administration of a single dosage form containing all components, or administration of separate dosage forms at substantially the same time. Sequential administration preferably includes administration according to different schedules such that there is an overlap in the period during which the composition useful herein and the other therapeutic agent are provided. Examples of other therapeutic agents include at least one microorganism of a different species or strain, a methanogen or a vaccine that inhibits methane production, and / or a methane production inhibitor, such as natural or chemically synthesized methane production inhibitor and / or bromoform.

[0264] Suitable agents with which the compositions useful herein may be administered separately, simultaneously, or sequentially include one or more prebiotic agents, one or more probiotic agents, one or more postbiotic agents, one or more phospholipids, one or more gangliosides, other suitable agents known in the art, and combinations thereof.

[0265] Typically, the term prebiotics refers to substances that stimulate the growth and / or activity of biologically active bacteria in the digestive system of animals. Prebiotics can be selectively fermented ingredients that allow specific changes in both the composition and / or activity of the gastrointestinal microflora, conferring health benefits to the host. Probiotics generally refer to microorganisms that contribute to the balance of gut microorganisms, which play a role in maintaining health or providing other biological activities. Many species of lactic acid bacteria (LAB), such as Lacticaseibacillus and Bifidobacterium, are generally considered probiotics, although some species of Bacillus and some yeasts have also been found to be suitable candidates. Postbiotics refer to non-viable bacterial products or metabolic by-products from microorganisms, such as probiotics, that have biological activity in the host.

[0266] Useful prebiotics include galactooligosaccharides (GOS), short-chain GOS, long-chain GOS, fructooligosaccharides (FOS), short-chain FOS, long-chain FOS, inulin, galactan, fructan, lactulose, and any mixture of any two or more thereof. Some prebiotics are reviewed in Boehm G and Moro G (Structural and Functional Aspects of Prebiotics Used in Infant Nutrition, J. Nutr. (2008) 138(9):1818S-1828S), incorporated herein by reference. Other useful agents may include dietary fiber, such as fully or partially insoluble or indigestible dietary fiber.

[0267] Thus, in one embodiment, L. rhamnosus strain FNZ129 and / or its derivatives may be administered separately, simultaneously or sequentially with one or more agents selected from one or more probiotics, one or more prebiotics, one or more dietary fiber sources, one or more galactooligosaccharides, one or more short chain galactooligosaccharides, one or more long chain galactooligosaccharides, one or more fructooligosaccharides, one or more short chain galactooligosaccharides, one or more long chain galactooligosaccharides, inulin, one or more galactans, one or more fructans, lactulose, or a mixture of any two or more thereof.

[0268] In certain embodiments, the composition comprises L. rhamnosus strain FNZ129 and / or a derivative thereof, and one or more prebiotics, one or more probiotics, one or more postbiotics, and one or more dietary fiber sources. In certain embodiments, the prebiotics comprise one or more fructooligosaccharides, one or more galactooligosaccharides, inulin, one or more galactans, one or more fructans, lactulose, or any mixture of any two or more thereof.

[0269] Without wishing to be bound by theory, it is believed that co-cultivation and / or simultaneous administration of two or more strains of lactic acid bacteria, such as three strains of lactic acid bacteria, can reduce the incidence of culture failure due to infection with bacteriophages. Thus, in a particular embodiment, the composition comprises L. rhamnosus FNZ129 and one or more other strains of lactic acid bacteria, preferably two or more other strains of lactic acid bacteria. In another embodiment, the composition comprising L. rhamnosus FNZ129 is administered simultaneously or sequentially with one or more other compositions comprising one or more other strains of lactic acid bacteria, preferably two or more other strains of lactic acid bacteria.

[0270] It is understood that different compositions of the present invention may be formulated with a view to administration to a particular subject group, for example, a particular ruminant subject group or a particular monogastric subject group. For example, a formulation of a composition suitable for administration to a cow may be different from that suitable for administration to a different ruminant animal, such as a sheep, and a formulation of a composition suitable for administration to a pig may be different from that suitable for administration to a different monogastric animal, such as a horse. It should also be recognized that the compositions of the present invention may be formulated differently to be suitable for administration to animals of different ages. For example, a formulation of a composition suitable for administration to a calf, a lamb, a piglet, or a foal may be different from that suitable for administration to an adult dairy cow, a sheep, a pig, or a horse. In certain embodiments, a first composition may be formulated for administration to a young animal, such as a pre-weaned animal, in an initial administration regimen, and a second composition may be formulated for administration to the same animal in a maintenance administration regimen. In some embodiments, a first composition is formulated for a pre-weaned animal and a second composition is formulated for a post-weaned animal.

[0271] Preparation of L. rhamnosus FNZ129 strain Direct-fed microorganisms (DFM) and their use in methods for regulating ruminal function and improving ruminant performance are known in the art, as are methods for their production.

[0272] Briefly, the L. rhamnosus FNZ129 strain can be cultured using conventional liquid or solid fermentation techniques. In at least one embodiment, the strain is grown in liquid nutrient broth to a level where the highest number of cells are formed. The strain is produced by fermenting a bacterial strain that can be initiated by scaling up a seed culture. This involves repeatedly and aseptically transferring the culture to increasingly larger volumes to serve as inoculum for fermentation, which can be carried out in large stainless steel fermenters in a medium containing the proteins, carbohydrates, and minerals necessary for optimal growth. Non-limiting exemplary media are MRS or TSB. However, other media can be used. After the inoculum is added to the fermentation vessel, the temperature and agitation are controlled to allow maximum growth. Once the culture reaches maximum population density, the culture is harvested by separating the cells from the fermentation medium. This is usually done by centrifugation.

[0273] In one embodiment, to prepare L. rhamnosus strain FNZ129, the strain is added at about 1×10 8 CFU / ml ~ approx. 1 x 10 9 The bacteria are fermented to a concentration of 1000 CFU / ml. The bacteria are harvested by centrifugation and the supernatant removed. The pelleted bacteria can be used to make a DFM. In at least some embodiments, the pelleted bacteria are lyophilized and then used to form a DFM. However, it is not necessary to freeze-dry the strain prior to use. The strain can be used with or without preservatives and in concentrated, non-concentrated, or diluted form.

[0274] The number of cultures can then be determined. CFU or colony forming units are the viable cell count of a sample obtained from a standard microbiological plating method. The term derives from the fact that a single cell, when plated on an appropriate medium, will grow into a viable colony in the agar medium.

[0275] The term colony forming unit is a more useful unit measurement than cell number because many cells can give rise to one visible colony.

[0276] Specific Embodiments Specific embodiments of the invention are described in the following numbered paragraphs: A1. The isolated Lacticase ibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. A2. A food or feed composition comprising Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated 2 August 2021) or a derivative thereof. A3. A food or feed composition according to A2, wherein the composition is a ruminant feed composition. A4. A ruminant feed composition for inhibiting the growth of methanogenic bacteria and / or archaea in the forestomach of a ruminant, reducing the ability of the rumen microflora to produce methane, reducing methane emissions by a ruminant, increasing feed efficiency in a ruminant, increasing growth and / or productivity in a ruminant, increasing the yield of milk and / or milk components produced from the ruminant, or improving the body weight and / or body composition of a ruminant, wherein the feed composition comprises Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. A5. The ruminant feed composition of A4, wherein the feed composition is a fermented yogurt-style composition, the fermented yogurt-style composition being formed through a process of growing L. rhamnosus FNZ129 using a milk-based carrier or a non-dairy-based carrier.

[0277] A6. A ruminant feed composition as described in A4 which is or comprises partial or total mixed ration (TMR), corn, soybean, forage, cereals, distillers grains, germinated grains, legumes, fiber, forage, grass, hay, straw, silage, grain, leaves, meal, mash, lick block, or molasses. A7. A ruminant feed composition according to any one of A4 to A6, further comprising at least one microorganism of a different species or strain, a methanogen or a vaccine that inhibits methane production, and / or a natural or chemically synthetic methane production inhibitor, and / or a methane production inhibitor such as bromoform. A8. A ruminant feed composition according to any one of A4 to A7, further comprising one or more agents selected from one or more prebiotics, one or more probiotics, one or more postbiotics, one or more dietary fiber sources, one or more galactooligosaccharides, one or more short chain galactooligosaccharides, one or more long chain galactooligosaccharides, one or more fructooligosaccharides, inulin, one or more galactans, one or more fructans, lactulose, or a mixture of any two or more thereof. A9. The ruminant feed composition of any one of A4 to A8, wherein the derivative of L. rhamnosus FNZ129 is a cell lysate of the strain, a cell suspension of the strain, a metabolic product of the strain, a culture supernatant of the strain, or killed L. rhamnosus FNZ129. A10. A method for inhibiting the growth of methanogenic bacteria and / or archaea in the forestomach of a ruminant comprising the step of administering to said animal a food or feed composition as defined in A2 or A3, or a ruminant feed composition as defined in any one of A4 to A9.

[0278] A11. A method for reducing the methane producing capacity of the ruminant animal's ruminal microflora, comprising administering to said animal a food or feed composition as defined in A2 or A3, or a ruminant feed composition as defined in any one of A4 to A9. A12. A method for reducing methane emission by a ruminant animal, comprising administering to said animal a food or feed composition as defined in A2 or A3, or a ruminant feed composition as defined in any one of A4 to A9. A13. A method for increasing feed efficiency in a ruminant animal, comprising the step of administering to said animal a food or feed composition as defined in A2 or A3, or a ruminant feed composition as defined in any one of A4 to A9. A14. A method for increasing the growth and / or productivity of a ruminant comprising administering to said animal a food or feed composition as defined in A2 or A3, or a ruminant feed composition as defined in any one of A4 to A9. A15. A method for increasing the yield of milk and / or milk components produced by a ruminant animal, comprising the step of administering to said animal a food or feed composition as defined in A2 or A3, or a ruminant feed composition as defined in any one of A4 to A9.

[0279] A16. A method for improving the body weight and / or body composition of a ruminant comprising administering to said animal a food or feed composition as defined in A2 or A3, or a ruminant feed composition as defined in any one of A4 to A9. A17. A method for inhibiting the growth of methanogenic bacteria and / or archaea in the forestomach of a ruminant, comprising administering to a monogastric animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. A18. A method for reducing methane emissions by a ruminant animal, comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. A19. A method for increasing feed efficiency in a ruminant animal, comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. A20. A method for increasing growth and / or productivity in a ruminant animal, comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0280] A21. A method for increasing the yield of milk and / or milk components produced by a ruminant animal, comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. A22. A method for improving the body weight and / or body composition of a ruminant comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. A23. A method for reducing the ability of the ruminal microflora to produce methane, comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. A24. The method according to any one of A17 to A23, wherein the growth of methylotrophic methanogens, preferably methanogens from the genus Methanosphaera, is inhibited in the forestomach of an animal. A25. The method of any one of A17 to A24, wherein L. rhamnosus FNZ129 or a derivative thereof is administered in a composition which is a food, beverage, food additive, beverage additive, animal feed, animal feed additive, animal feed additive, animal feed supplement, dietary supplement, carrier, vitamin or mineral premix, nutritional product, enteral nutritional product, soluble, supplement, pharmaceutical, lick block, drench, tablet, capsule, pellet or intraluminal product, e.g., a bolus, or L. rhamnosus FNZ129 is encapsulated, e.g., in a liposome, microbubble, microparticle or microcapsule.

[0281] A method according to A25, wherein L. rhamnosus FNZ129 or a derivative thereof is administered in drinking water, milk, milk powder, milk replacer, milk fortifier, whey, milk powder, partial or total mixed ration (TMR), corn, soybean, feed, cereals, distillers grains, germinated cereals, legumes, vitamins, leaves, minerals, fibre, forage, grass, hay, straw, silage, grains, leaves, meal, solubles, supplements, mash feed, meal, fruit pulp, vegetable pulp, fruit or vegetable pomace, citrus meal, wheat shorts, corn cob meal, molasses, sucrose, maltodextrin, rice husk, vermiculite, zeolite or ground limestone. A27. 10 per kg dry weight carrier feed 4 ~10 13 Colony forming units, 10 per kg of animal weight per day 4 ~10 10 colony forming units, or 10 per day 4 ~10 13 The method of any one of A17 to A26, comprising administering the colony forming units to the animal. A28. 10 per kg dry weight carrier feed 8 ~10 12 Colony forming units, 10 per kg of animal weight per day 5 ~10 8 colony forming units, or 10 per day 6 ~10 13The method of A27, comprising administering the colony forming units to the animal. A29. The method of any one of A17 to A28, wherein the derivative of L. rhamnosus FNZ129 is a cell lysate of the strain, a cell suspension of the strain, a metabolic product of the strain, a culture supernatant of the strain, or killed L. rhamnosus FNZ129. A30. The method according to any one of A17 to A29, further comprising administering at least one microorganism of a different species or strain, a methanogen or a vaccine that inhibits methane production, and / or a natural or chemically synthetic methane production inhibitor and / or a methane production inhibitor such as bromoform.

[0282] A31. The method of any one of A17 to A30, wherein L. rhamnosus FNZ129 or a derivative thereof is administered separately, simultaneously or sequentially with one or more agents selected from one or more prebiotics, one or more probiotics, one or more postbiotics, one or more dietary fiber sources, one or more galactooligosaccharides, one or more short chain galactooligosaccharides, one or more long chain galactooligosaccharides, one or more fructooligosaccharides, inulin, one or more galactans, one or more fructans, lactulose, or a mixture of any two or more thereof. A32. A method according to any one of A17 to A31, which enhances growth or productivity of a ruminant. A33. The method according to A32, which increases the yield of milk and / or milk components produced from a ruminant animal. A34. The method according to A33, which increases the yield of milk fat, milk protein or milk solids in milk produced from a ruminant. A35. The method according to any one of A17 to A34, further improving the body weight and / or body composition of a ruminant.

[0283] A36. The method according to any one of A17 to A35, wherein the ruminant is a cow, goat, sheep, bison, yak, buffalo, deer, camel, alpaca, llama, wildebeest, antelope, or nilgai. A37. The method according to any one of A17 to A36, wherein the ruminant is a cow or a sheep. A38. The method according to any one of A17 to A37, wherein the ruminant is a cow. A39. The method according to any one of A17 to A38, wherein the ruminant is a lactating animal. A40. The method according to any one of A17 to A38, wherein the ruminant is a pre-weaned animal, such as a calf or a lamb.

[0284] A41. The method according to any one of A17 to A38, wherein the ruminant is a weaned animal. A42. The method of any one of A17-A38, wherein L. rhamnosus FNZ129 is administered to the ruminant both pre-weaning and post-weaning. A43. The method according to any one of A17 to A38, wherein administration is to a pre-weaned animal and the inhibition of the growth of methanogenic bacteria and / or archaea in the forestomach of the ruminant, the reduction in methane emission by the ruminant, e.g. methane production, and / or the increased feed efficiency in the ruminant persists after weaning. A44. The method of any one of A17 to A43, wherein the inhibition of the growth of methanogenic bacteria and / or archaea in the forestomach of the ruminant, the reduction in methane emission by the ruminant, e.g., methane production, and / or the increase in feed efficiency in the ruminant persists for at least 2 days, 3 days, 5 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, or 7 years after the final administration of L. rhamnosus FNZ129. A45. The method according to A44, wherein the inhibition of the growth of methanogenic bacteria and / or archaea in the forestomach of the ruminant, the reduction in methane emission by the ruminant, e.g. methane production, and / or the increase in feed efficiency in the ruminant persists throughout the life of the ruminant.

[0285] A46. Use of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated 2 August 2021) or a derivative thereof in the manufacture of a composition for inhibiting the growth of methanogens and / or archaea in the forestomach of a ruminant, reducing the methane-producing capacity of the rumen microflora, reducing methane emissions by a ruminant, increasing feed efficiency in a ruminant, increasing the yield of milk and / or milk components produced from a ruminant, or improving the body weight and / or body composition of a ruminant. A47. The use according to A46, wherein the composition comprises a food or feed composition as defined in A2 or A3, or a ruminant feed composition as defined in any one of A4 to A9. A48. Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated 2 August 2021) or a derivative thereof for use in inhibiting the growth of methanogens and / or archaea in the forestomach of a ruminant, reducing the methane-producing capacity of the rumen microflora, reducing methane emissions by a ruminant, increasing feed efficiency in a ruminant, increasing the yield of milk and / or milk components produced by a ruminant, or improving the body weight and / or body composition of a ruminant.

[0286] B1. Isolated Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. B2. A food or feed composition comprising Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. B3. A food or feed composition according to B2, which is a feed composition for a monogastric animal. B4. A monogastric feed composition for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of a monogastric animal, reducing the methane producing capacity of the gastrointestinal microflora, reducing methane emissions by the monogastric animal, increasing feed efficiency in a monogastric animal, enhancing growth and / or productivity in a monogastric animal, increasing the yield of milk and / or milk components produced from the monogastric animal, or improving the body weight and / or body composition of a monogastric animal, the feed composition comprising Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. B5. The monogastric feed composition of B4, wherein the feed composition is a fermented yogurt-style composition, the fermented yogurt-style composition being formed through a process of growing L. rhamnosus FNZ129 using a dairy-based carrier or a non-dairy-based carrier.

[0287] B6. The monogastric feed composition according to B4, which is or comprises partial or complete mixed ration (TMR), corn, soybean, forage, cereals, distillers grains, germinated grains, legumes, fiber, forage, grass, hay, straw, silage, grain, leaves, meal, mash feed, fruit pulp, vegetable pulp, fruit or vegetable pomace, citrus meal, wheat shorts, corn cob meal, or molasses. B7. A monogastric feed composition according to any one of B4 to B6, further comprising at least one microorganism of a different species or strain, a methanogen or a vaccine that inhibits methane production, and / or a natural or chemically synthetic methane production inhibitor, and / or a methane production inhibitor such as bromoform. B8. The monogastric feed composition according to any one of B4 to B7, further comprising one or more agents selected from one or more prebiotics, one or more probiotics, one or more postbiotics, one or more dietary fiber sources, one or more galactooligosaccharides, one or more short chain galactooligosaccharides, one or more long chain galactooligosaccharides, one or more fructooligosaccharides, inulin, one or more galactans, one or more fructans, lactulose, or a mixture of any two or more thereof. B9. The monogastric feed composition according to any one of B4 to B8, wherein the derivative of L. rhamnosus FNZ129 is a cell lysate of the strain, a cell suspension of the strain, a metabolic product of the strain, a culture supernatant of the strain, or killed L. rhamnosus FNZ129. B10. A method for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of a monogastric animal, comprising the step of administering to said animal a food or feed composition as defined in B2 or B3, or a monogastric feed composition as defined in any one of B4 to B9.

[0288] B11. A method for reducing the ability of the gastrointestinal microflora of a monogastric animal to produce methane, comprising administering to said animal a food or feed composition as defined in B2 or B3, or a monogastric feed composition as defined in any one of B4 to B9. B12. A method for reducing methane emission by a monogastric animal, comprising administering to said animal a food or feed composition as defined in B2 or B3, or a monogastric feed composition as defined in any one of B4 to B9. B13. A method for increasing feed efficiency in a monogastric animal, comprising administering to said animal a food or feed composition as defined in B2 or B3, or a monogastric feed composition as defined in any one of B4 to B9. B14. A method for increasing the growth and / or productivity of a monogastric animal, comprising administering to said animal a food or feed composition as defined in B2 or B3, or a monogastric feed composition as defined in any one of B4 to B9. B15. A method for increasing the yield of milk and / or milk components produced by a monogastric animal, comprising the step of administering to said animal a food or feed composition as defined in B2 or B3, or a monogastric feed composition as defined in any one of B4 to B9.

[0289] B16. A method for improving the body weight and / or body composition of a monogastric animal, comprising administering to said animal a food or feed composition as defined in B2 or B3, or a monogastric feed composition as defined in any one of B4 to B9. B17. A method for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of a monogastric animal, comprising administering to the monogastric animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. B18. A method for reducing methane emission by a monogastric animal, comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. B19. A method for increasing feed efficiency in a monogastric animal, comprising administering to the animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. B20. A method for reducing the ability of gastrointestinal microflora to produce methane, comprising administering to an animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

[0290] B21. A method for increasing growth and / or productivity in a monogastric animal, comprising administering to the monogastric animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. B22. A method for improving body weight and / or body composition in a monogastric animal, comprising administering to the monogastric animal an effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. B23. The method according to any one of B17 to B22, wherein the growth of methylotrophic methanogens, preferably methanogens from the genus Methanosphaera, is inhibited in the caecum of an animal. B24. The method of any one of B17 to B23, wherein L. rhamnosus FNZ129 or a derivative thereof is administered in a composition which is a food, beverage, food additive, beverage additive, animal feed, animal feed additive, animal feed supplement, nutritional supplement, carrier, vitamin or mineral premix, nutritional product, enteral nutritional product, soluble, slurry, supplement, pharmaceutical, lick block, drench, tablet, capsule, pellet or bolus, or L. rhamnosus FNZ129 is encapsulated, for example, in a liposome, microbubble, microparticle or microcapsule. The method according to claim 24, wherein L. rhamnosus FNZ129 or a derivative thereof is administered in drinking water, milk, milk powder, milk replacer, milk fortifier, whey, whey powder, feed pellets, corn, soybeans, feed, cereals, distillers grains, germinated grains, legumes, vitamins, amino acids, minerals, fibre, feed, grass, hay, silage, grains, leaves, meal, solubles, slurry, supplements, mash feed, meal, fruit pulp, vegetable pulp, fruit or vegetable pomace, citrus meal, wheat shorts, corn cob meal, molasses, sucrose, maltodextrin, rice husk, vermiculite, zeolite or ground limestone.

[0291] B26. 10 per kg dry weight carrier feed 4 ~10 13 Colony forming units, 10 per kg of animal weight per day 4 ~10 10 colony forming units, or 10 per day 4 ~10 13 The method of any one of B17 to B25, comprising administering the colony forming units to the animal. B27. 10 per kg dry weight carrier feed 8 ~10 12 Colony forming units, 10 per kg of animal weight per day 5 ~10 8 colony forming units, or 10 per day 6 ~10 13 The method of B26, comprising administering the colony forming units to the animal. B28. The method according to any one of B17 to B27, wherein the derivative of L. rhamnosus FNZ129 is a cell lysate of the strain, a cell suspension of the strain, a metabolic product of the strain, a culture supernatant of the strain, or killed L. rhamnosus FNZ129. B29. The method according to any one of B17 to B28, further comprising administering at least one additional microorganism of a different species or strain, a methanogen or a vaccine that inhibits methane production, and / or a methane production inhibitor, such as a natural or chemically synthetic methane production inhibitor and / or bromoform. B30. The method according to any one of B17 to B29, wherein L. rhamnosus FNZ129 or a derivative thereof is administered separately, simultaneously or sequentially with one or more agents selected from one or more prebiotics, one or more probiotics, one or more postbiotics, one or more sources of dietary fiber, one or more galactooligosaccharides, one or more short chain galactooligosaccharides, one or more long chain galactooligosaccharides, one or more fructooligosaccharides, inulin, one or more galactans, one or more fructans, lactulose, or a mixture of any two or more thereof.

[0292] B31. The method of any one of B17-B30, additionally improving body weight and / or body composition in a monogastric animal. B32. The method of any one of B17-B31, wherein the monogastric animal is a human, pig, cat, dog, horse, donkey, rabbit, or poultry. B33. The method of any one of B17-B31, wherein said monogastric animal is a companion animal. B34. The method of any one of B17 to B31, wherein said monogastric animal is a non-human animal. B35. The method of any one of B17-B31, wherein said monogastric animal is a pig.

[0293] B36. The method of any one of B17-B31, wherein the monogastric animal is a chicken, duck, goose, or turkey. B37. The method of any one of B17-B35, wherein the monogastric animal is a pre-weaned animal, such as a piglet or a foal. B38. The method of any one of B17-B35, wherein the monogastric animal is a post-weaned animal. B39. The method of any one of B17 to B35, wherein L. rhamnosus FNZ129 is administered to the monogastric animal both pre-weaning and post-weaning. B40. The method according to any one of B17 to B35, wherein administration is to a pre-weaned animal and inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of the monogastric animal, reducing methane emission, e.g. methane production, by the monogastric animal, and / or increasing feed efficiency in the monogastric animal persists after weaning.

[0294] B41. The method of any one of B17 to B35, wherein the inhibition of the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of the monogastric animal, the reduction in methane emission, e.g., methane production, by the monogastric animal, and / or the increase in feed efficiency in the monogastric animal persists for at least 2 days, 3 days, 5 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, or 7 years after the final administration of L. rhamnosus FNZ129. B42. The method of B41, wherein the inhibition of the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of the monogastric animal, the reduction in methane emission, e.g., methane production, by the monogastric animal, and / or the increase in feed efficiency in the monogastric animal persists throughout the life of the monogastric animal. B43. The method of any one of B17 to B42, wherein L. rhamnosus FNZ129 is administered in a composition that is a fermented yogurt-style composition, the fermented yogurt-style composition being formed through a process of growing L. rhamnosus FNZ129 using a milk-based carrier. B44. Use of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof in the manufacture of a composition for inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of a monogastric animal, reducing the methane production potential of the gastrointestinal microflora, reducing methane emissions by a monogastric animal, increasing feed efficiency in a monogastric animal, improving growth and / or productivity in a monogastric animal, increasing the yield of milk and / or milk components produced from a monogastric animal, or improving the body weight and / or body composition of a monogastric animal. B45. The use according to B44, wherein the composition is a medicament.

[0295] B46. The use of B44 or B45, wherein the monogastric animal is a human. B47. Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof for use in inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of a monogastric animal, reducing the methane production capacity of the gastrointestinal microflora, reducing methane emission by a monogastric animal, increasing feed efficiency in a monogastric animal, improving growth and / or productivity in a monogastric animal, increasing the yield of milk and / or milk components produced by a monogastric animal, or improving the body weight and / or body composition of a monogastric animal. B48. L. rhamnosus FNZ129 or a derivative thereof for use in B47, wherein the monogastric animal is a human. EXAMPLES

[0296] 1. Example 1 - Plate-based screening of bacteriocin extracts against indicator methanogenic strains 1.1 Materials and Methods 1.1.1 Bacteriocin extraction Bacteriocin extracts from L. rhamnosus FNZ129 cultures were prepared and tested for their effect on indicator methanogenic strains Methanobrevibacter boviskoreani JH1 ("JH1"), Methanosphaera sp. WGK6 ("WGK6"), Methanobrevibacter ruminantium M1 ("M1") and Methanobrevibacter gottschalkii D5 ("D5").

[0297] L. rhamnosus FNZ129 was revived from -80°C storage by plating on De Man-Rogosa-Sharpe agar (MRS, De Man et al., 1960) + lactose (2 g / L). Using a small inoculation loop, the glycerol stock was streaked onto MRS agar plates to obtain isolated colonies. The plates were incubated at 37°C for 48 h in a sealed container. After growth, a single colony was selected, picked up, restreaked onto an agar plate, and incubated at 37°C to obtain isolated colonies. After 48 h, a single colony was selected from the restreaked plate, inoculated into MRS liquid medium, and incubated at 37°C for 48 h. An inoculum (1 mL) of each revived strain was then subcultured into 16 mL of MRS + nisin liquid medium (1 ng / mL final concentration). Nisin was included in these media at a very low level to induce bacteriocin production. The cultures were incubated overnight at 37°C. An overnight grown L. rhamnosus FNZ129 culture was used for bacteriocin extraction. A drop of the culture was used to make a wet mount slide, the cells were examined using a phase contrast microscope, and a Gram stain was prepared to check the purity of the culture.

[0298] The remainder of the culture (approximately 16 mL) was transferred to a 50 mL Falcon tube and used for bacteriocin extraction according to the method of Gaspar et al. (2018) with some modifications as follows: The pH of the culture was adjusted to approximately 6.8 with 6 M NaOH. 0.3 mL of catalase (2 mg / L) was then added to the culture and incubated at 37 °C for 30 min, followed by incubation at 70 °C for 45 min. The culture was then centrifuged at 12,000 × g for 20 min at 4 °C, the supernatant was decanted, and the cell pellet was resuspended in 8 mL of 0.9% NaCl, pH 2. The pH of the resuspended pellet was checked and, if necessary, adjusted to pH 2 with 1 M HCl. The cells were incubated for 2 h at 4 °C with gentle agitation on a shaking platform. The cells were then centrifuged at 12,000 × g for 20 min at 4 °C, and the supernatant was collected in a new 15 mL Falcon tube. The pH of the supernatant was adjusted to pH 6.8 with 1 M NaOH and filtered under sterile conditions through a sterile filter (Millex-GP 0.22 μm, 25 mm diameter, Millipore, Merck, Sigma-Aldrich, New Zealand) using a 10-mL syringe and needle into a sterile Hungate tube flushed with N. The filtered supernatant was frozen at −20° C. until use.

[0299] 1.1.2 Mbb.boviskoreani JH1 culture To identify potential candidate LAB strains with anti-methanogen activity, we used a microtiter plate-based methanogen growth inhibition bioassay using the model methanogen strain Methanobrevibacter boviskoreani JH1 (Li et al., 2019). Mbb. boviskoreani JH1 has the unique ability to grow using ethanol as a reducing power source to reduce CO2 to CH4, and JH1 was grown in microtiter plates and incubated under anaerobic conditions without the need for H2 supplementation with 1 atm H2:CO2 overpressure (80:20). This allows a high-throughput JH1 screening method to identify inhibitory activity from LAB strains.

[0300] Mbb. boviskoreani JH1 cultures for inoculating plate assays were grown by syringe using anaerobic techniques in Balch tubes (anaerobic tubes, 18 × 150 mm, butyl rubber septum stoppers, aluminum crimp, Bellco Glass, Vineland, NJ, USA) containing 9 mL of BY medium (Joblin, 1995) supplemented with (final concentrations) 60 mM sodium formate, 200 mM ethanol, 0.1 mL of vitamin solution (1×) and 0.1 mL of coenzyme M solution (10 μM). The tubes were incubated at 39 °C without shaking until visible turbidity appeared after 3–5 days and were used to inoculate the microtiter plate assays after reaching an OD600 of 0.8–1.0 against a distilled water blank. Overpressure in the JH1 culture tubes was released by inserting a needle into the butyl rubber septum and allowing accumulated gas to escape before removing the inoculum.

[0301] Freshly grown cultures were checked using wet mounts and Mbb. boviskoreani JH1 under a fluorescent microscope, where Mbb. boviskoreani JH1 appeared as short oval rods fluorescing green under ultraviolet (UV) illumination. Contamination of the cultures was checked by inoculating a sample of the culture into 9 mL of BY medium supplemented with 5 mM glucose and incubating at 39 °C for 1 day. If no turbidity was observed after 1 day, the culture was considered uncontaminated. Further culture validation was sometimes performed by extracting genomic DNA from the cultures and PCR amplifying the 16S rRNA gene using both regular bacterial 16S primers (27f-GAGTTTGATCMTGGCTCAG, 1492r-GGYTACCTTGTTACGACTT) and archaeal specific 16S primers (915af-AGGAATTGGCGGGGGAGCAC, 386r-GCGGTGTGTGCAAGGAGC). Culture purity was confirmed using the presence of bands with the archaeal primer set and the absence of bands with the bacterial primer set, as well as sequencing results from the PCR products.

[0302] 1.1.3 Cultivation of Methanosphaera sp. WGK6 Members of the genus Methanosphaera account for approximately 8% of ruminal methanogens (Henderson et al., 2015) and are generally H2-dependent methylotrophs that reduce methanol to methane. Methanosphaera sp. WGK6 is an H2-utilizing methylotrophic methanogen isolated from an Australian kangaroo, but can also utilize ethanol as a source of reducing power to reduce methanol to methane (Hoedt, 2017). Similar to Mbb. boviskoreani JH1, this metabolic capability theoretically allows WGK6 to grow on ethanol without the need for H2 overpressure and thus to grow in microtiter plates. Growth of Methanosphaera sp. WGK6 was achieved in 10-mL cultures on BRN-RF10 medium (Balch et al., 1979; Hoedt, 2016). WGK6 was tested using either H2 (180 kPa overpressure of H2+CO2, 80:20) or ethanol as the energy source and terminal electron acceptor, in both cases with ethanol as the terminal electron acceptor, using a pressurizable stainless steel gas canister (Bernards 2017). Attempts to grow WGK6 on ethanol+methanol were unsuccessful, but WGK6 could be grown on methanol+H2 in a Hungate tube. Initial attempts to culture Methanosphaera sp. WGK6 in a microtiter plate format in a pressurizable gas canister with H2+CO2 atmosphere (180 kilopascal overpressure) produced barely detectable growth after one week. However, good growth of Methanosphaera sp. WGK6 was obtained after increasing concentrations of cysteine ​​added to the BRN-RF10 medium. Therefore, a plate assay using a pressurizable stainless steel gas canister (H2+CO2; 80:20) was developed.

[0303] Methanosfera species WGK6 cultures for analysis were grown in Balch tubes in 9 mL of BRN-RF10 medium supplemented with 60 mM sodium formate, 1% methanol, 0.1 mL of vitamin solution (1x) and 0.1 mL of coenzyme M solution (10 µM) using anaerobic techniques and 180 kPa overpressure of H2+CO2 (80:20, BOC Gas, New Zealand) by syringe. The tubes were incubated at 39 °C for 3–5 days without shaking until visible turbidity appeared and after reaching an OD600 of 0.8–1.0 against a distilled water blank, they were used for inoculation of the test tube assays. The overpressure in the WGK6 culture tubes was released by inserting a needle into the butyl rubber septum and allowing the accumulated gas to escape before removing the inoculum.

[0304] 1.1.4 Mbb.ruminantium M1 and Mbb.gottschalkii D5 culture The procedure for growing Methanobrevibacter ruminantium M1 and Methanobrevibacter gottschalkii D5 was identical to the WGK6 protocol described in 1.1.3 above, except that BY medium was used for growth. Cultures for the assay were grown in 9 mL of BY medium supplemented with 60 mM sodium formate, 0.1 mL of vitamin solution (1x) and 0.1 mL of coenzyme M solution (10 μM) added by syringe in Balti tubes, using anaerobic techniques and 180 kPa overpressure of H2+CO2 (80:20, BOC Gas, New Zealand). Tubes were incubated at 39 °C without shaking until visible turbidity appeared after 3–5 days and used to inoculate the microtiter plate assays.

[0305] 1.1.5 Mbb.boviskoreani JH1 growth inhibition assay Bacteriocin extracts from L. rhamnosus FNZ129 stored frozen under anaerobic conditions in Hungate tubes were allowed to thaw at room temperature. All assay components for each assay, except the JH1 inoculum, were added to 3.75 mL of BY+formic acid medium in sterile 7.5 mL Hungate tubes via CO2-flushed syringes and needles in the proportions shown in Table 1. Each test tube was then inoculated with a freshly grown JH1 culture and incubated at 39°C for 1 h before being moved into an anaerobic chamber (98% CO2-2% H2 atmosphere; Coy Laboratory Products, USA) and dispensed into the wells of a multi-well 96-well plate. The filled plate was placed into an AnaeroPack 2.5 L rectangular jar with MCG Anaeropack-Anaero Ngaio Diagnostics, Nelson, NZ), the lid was sealed, and the jar was removed from the anaerobic chamber and incubated at 39°C. Plates were observed daily through transparent jars until Mbb. boviskoreani JH1 control wells showed visible turbidity (usually within 5-6 days). The optical density of each well was then recorded at 595 nm (OD595) after shaking for 5 s in a Multiscan FC microplate photometer (Thermo Scientific, Auckland, New Zealand). The absorbance readings of the medium control wells were subtracted as background and the % inhibition of Mbb was subtracted. Boviskoreani JH1 growth caused by the bacteriocin extract samples was calculated relative to the JH1 positive growth control wells (which contained buffer only).

[0306] [Table 1]

[0307] 1.1.6 Methanosphaera sp. WGK6 growth inhibition assay Each assay component of the assay, except for the WGK6 inoculum, was added via a CO2 flush syringe and needle to 3.75 mL of BRN-RF10 medium in a Hangert tube supplemented with 1% methanol (247 mM, final concentration), as described in Table 4. The tube along with the inoculation tube was then moved into the chamber. The medium containing all components except the inoculum was dispensed into the plates in the chamber, and the inoculum was then added to the appropriate wells. The plates were placed in a stainless steel gas canister laid horizontally to hold up to four microtiter plates at a time. Two anaerobic pouches (MCG Anaeropack-Anaero, Ngaio Diagnostics, Nelson, NZ) were added, the canister was sealed, and the canister was removed from the anaerobic chamber, pumped to 180 kPa with H2+CO2 (80:20, BOC Gases, NZ), and then incubated at 39°C for 1 week. The canister was checked periodically to ensure that overpressure was maintained and repressurized with H2+CO2 as necessary. After 1 week of incubation, the canister was opened and the plate removed. The contents of each well were evenly resuspended by repeated pipetting with a multichannel pipettor. The optical density of each well was then immediately recorded at 595 nanometers (OD595) after shaking for 5 seconds in a Multiscan FC microplate photometer (Thermo Scientific, Auckland, New Zealand). The absorbance readings of the medium control wells were subtracted as background to calculate the % inhibition of Methanosphaera sp. WGK6 growth caused by the bacteriocin extract samples relative to the WGK6 positive growth control wells (containing only buffer instead of the bacteriocin extract).

[0308] [Table 2]

[0309] 1.1.7 Mbb.ruminantium M1 and Mbb.gottschalkii D5 growth inhibition assay Cultures of Mbb. ruminantium M1 and Mbb. gottschalkii D5 were prepared as described above in 1.1.4. Before removing the inoculum, the overpressure in the tube was released.

[0310] The assay components were added to 3.5 mL of sterile BY medium in 7.5 mL Hungert tubes via CO2 flushed syringes and needles as described in Table 3. Each tube was then inoculated with a freshly grown culture and incubated at 39°C for 1 hour, then transferred to an anaerobic chamber and dispensed into wells of a 96-well multiwell plate. The plates were sealed and incubated under 180 kilopascal overpressure of H2+CO2 in a stainless steel gas canister, and their optical densities were recorded spectrophotometrically at OD595 as described for the Methanospharea sp. WGK6 assay in 1.1.6 above. The OD595 readings of the BY medium control wells were subtracted as background, and the % inhibition of growth of Mbb. ruminantium M1 or Mbb. gottschalkii D5 caused by the bacteriocin extract samples was calculated compared to the positive growth control wells (which contained buffer instead of the bacteriocin extract).

[0311] [Table 3]

[0312] 1.2 Results A total of 1,712 bacteriocin extracts from strains of lactic acid bacteria were screened against Methanosphaera spp. WGK6. Of these, 1,580 strains (>92%) showed less than 50% inhibition. The 1,712 strains of lactic acid bacteria included 94 strains of Lacticaseibacillus rhamnosus, of which 62 strains (~66%) showed less than 20% inhibition of WGK6, 81 strains (~86%) showed less than 50% inhibition, and only 3 strains (~3%) showed more than 80% inhibition. Taken together, this indicates that inhibition of methanogens is likely a strain-specific effect.

[0313] The L. rhamnosus FNZ129 bacteriocin extract showed very strong inhibition of the indicator methylotrophic methanogen Methanosphaera sp. WGK6, but no inhibition of the indicator hydrogenomethylenterans Mbb. boviskoreani JH1, Mbb. Ruminantium M1 or Mbb. gottschalkii D5, as shown in Table 4.

[0314] [Table 4]

[0315] 1.3 Discussion and Conclusions Members of the genera Methanobrevibacter and Methanosphaera are the major methanogens in the rumen across multiple ruminant species. WGK6 was used as an indicator strain for methylotrophic methanogens in general, and Methanosphaera species in particular. Mbb. boviskoreani JH1, Mbb. ruminantium M1, and Mbb. gottschalkii D5 were used as indicator strains for Methanobrevibacter species. This example shows that L. rhamnosus FNZ129 bacteriocin extract exhibits a strong inhibitory effect on the methylotrophic methanogen Methanosphaera species WGK6, but not on the hydrogenotrophic Mbb. boviskoreani JH1, Mbb. ruminantium M1, and Mbb. Gottschalkii D5 methanogens.

[0316] 2. Example 2 - Effect of L. rhamnosus FNZ129 on Ruminal In Vitro Assays 2.1 Materials and Methods 2.1.1 Preparation of bacterial cultures and test supernatants Seven Hungate tubes containing 5 mL of anaerobic MRS medium (Sigma-Aldrich) were inoculated with L. rhamnosus FNZ129 and grown at 39°C for 16 h (until the cultures reached stationary phase). The cultures were pooled in a 250-mL serum bottle flushed with CO2. A portion (1 mL) of the combined cultures was added to 9 mL of sterile MRS medium and its OD600 was measured. Further aliquots (0.5 mL) of the culture mixture were inoculated in triplicate into 4.5 mL of sterile anaerobic buffer, serially diluted 10-fold with CO2, plated on MRS plates, and the colony forming units (CFU / mL) of the original cultures were measured. Half of the remaining cultures were used for one set of ruminal in vitro fermentations (test cultures) and the other half was filtered (Millipore 0.22 μm pore size) and the filtrate was placed in a new sterile anaerobic serum bottle (supernatant treatment, SN). For the untreated control (buffer), untreated Lint's buffer (0.46 M K2HPO4; 0.54 M KH2PO4, pH 7) was used.

[0317] 2.1.1 Ruminal fluid preparation and in vitro fermentation setup For inoculation of the ruminal in vitro fermentation vessels, fresh ruminal contents were collected from six rumen-fistulated Friesian cows. After squeezing through one layer of cheesecloth, the ruminal fluid obtained from two animals was combined (approximately 150 mL of ruminal fluid) to obtain three biological replicates. An aliquot (12.5 mL) of the mixed ruminal fluid was added to 0.5 mg of dried grass and 36.5 mL of anaerobic phosphate buffer in a 250 mL serum bottle. Buffer, test culture, either SN or bacteriocin extract treatment (1 mL) was added, after which the serum bottle was closed with a butyl rubber stopper to obtain a final fermentation volume of 50 mL containing 25% ruminal fluid (v / v). Gas production and methane content were measured using an automated incubation system (Muetzel et al., 2014).

[0318] 2.1.2 VFA sample collection and analysis Samples were collected from the bottles for VFA analysis. At each time point, 3 mL aliquots were collected and their pH was measured. 1.8 mL samples of these aliquots were used for VFA and non-VFA analysis. VFA samples were centrifuged at 21,000×g for 10 min at 4° C., 0.9 mL of supernatant was removed and added to 0.1 mL of internal standard (20 mM 2-ethylbutyrate in 20% phosphoric acid), mixed, and frozen at −20° C. until analysis. After thawing and recentrifuging at 21,000×g for 10 min at 4° C., 0.9 mL was collected for derivatization for non-VFA analysis, while the remainder of the sample was analyzed directly by GC.

[0319] 2.2 Results L. rhamnosus FNZ129 was tested for its effect on gas production in a ruminal in vitro assay, as shown in Tables 5-10. Data shown are the average of three replicate experiments. Negative numbers represent stimulation, not inhibition. Asterisks (*) are used to indicate statistical significance (p<0.05) by Student's T-test with Welch's correction.

[0320] [Table 5]

[0321] [Table 6]

[0322] [Table 7]

[0323] [Table 8]

[0324] [Table 9]

[0325] [Table 10]

[0326] The addition of L. rhamnosus FNZ129 broth culture caused a decrease in the total gas produced, especially at 2 h, whereas the culture supernatant and extract showed little or no effect on the total gas produced. However, both the broth culture and the culture supernatant caused a significant decrease in the total amount of methane produced at 2 and 6 h.

[0327] While L. rhamnosus FNZ129 broth culture and culture supernatant showed little effect on the volatile fatty acids produced, the bacteriocin extract significantly stimulated the overall production of volatile fatty acids at 6 hours. This effect can also be seen as a significant stimulation of acetate and butyrate production at 6 hours, and a smaller stimulation of propionate production at 6 hours.

[0328] It should also be noted that ruminal in vitro studies are closed systems and nutrition may become limiting over time, therefore the 0-12 hour time point may more accurately reflect the in vivo situation, as animals typically ingest more food and liquid over a 24 hour period.

[0329] 2.3 Conclusion This example shows that L. rhamnosus FNZ129 showed a significant effect on fermentation end products. Overall, the results show that L. rhamnosus FNZ129 and its culture supernatant significantly reduced the total amount of methane produced in the ruminal in vitro model. This may be mediated by one or more compounds secreted by the bacteria into the culture supernatant. It also shows that the bacteriocin extract from FNZ129 has a significant stimulatory effect on the production of volatile fatty acids, especially acetate and butyrate. This suggests a shift in hydrogen metabolism from methane production to short-chain / volatile fatty acid (VFA) production and / or a disruption of intermediate cross-feeding between members of the microbiota due to changes in the ruminal microbiota. The significant increase in total VFA and butyrate, as well as the increase in propionate, suggests that animal feed efficiency is also likely to be improved.

[0330] 3. Example 3 - Effect of L. rhamnosus FNZ129 on piglet body weight 3.1 Materials and Methods The experimental protocol for the porcine probiotic study using a freeze-dried probiotic product (Fonterra) was approved by the AgResearch Grasslands Animal Care and Ethics Committee (approval number 15323).

[0331] A total of 16 piglets were enrolled in the study at 3 days of age. The piglets were weighed and randomly assigned to one of two treatment groups (n=8): 5×10 10CFU / day FNZ129 group; and a control group not receiving LAB feeding. Piglets were individually housed in custom-built cages configured to allow them to see, hear, and smell neighboring piglets while minimizing physical contact (as described in Fil et al., 2021), and each had free access to a heating pad, an automatic milk feeder, and water via a water bowl. Piglets remained in these cages except while the cages were being cleaned. However, piglets were kept in a large communal open enclosure where they could interact and play with each other for at least 2 h per day. For the first 5 weeks, piglets were fed only milk (reconstituted from powdered milk). To feed the piglets, the amount of milk required for 8 piglets was prepared, and one bag of freeze-dried (FD) FNZ129 containing the dose required for 8 animals was added to the milk. The milk was placed in an automatic milk feeder connected to an electronic system, allowing regular distribution of milk over a 24-h period. Frozen ice packs were placed around the feeder reservoir to keep the milk cool and prevent microbial growth. Pigs were weighed every 3 days.

[0332] At 5 weeks of age, solid food was introduced to the diet in the form of Little Pig Tucker pellets (NRM Feeds, NZ) and the piglets were weaned by slowly reducing the amount of milk offered (while keeping the probiotic dosage constant) until the 8th week of age, when only pellets were offered twice a day (morning; afternoon). Water was available ad libitum until the end of the study. During this period, at 7 weeks of age, the piglets (approximately 20 kg live weight; LWT) were moved to large pig pens in a covered barn. Each pen contained a raised wooden bedding area with a heating pad, a feeding trough, and water available via a self-actuating nipple. Once the piglets were fully transitioned onto the pellet diet, the amount of FNZ129 required for 8 pigs was reconstituted with a small amount of water (200 mL) and mixed uniformly into approximately 1.6 kg of pellets. The pellets containing FNZ129 were then divided equally into 8 aliquots (200 g / pig) and fed to the appropriate pigs. Control animals were fed the same amount of pellets treated with water only. These pellets + treatment mixtures were offered as the first meal in the morning when the pigs were hungry to ensure that the entire LAB dose was consumed each day. Once the pellets + treatment mixtures were eaten, a staple of dry pellets was supplemented for the morning meal. Pigs were fed ad libitum pellets from 8 weeks of age until the end of the study at 19 weeks of age.

[0333] The piglet experienced an episode of rotavirus infection in week 4 of the study and the animals were given electrolyte therapy and Scourban Plus (Bayer NZ). A second rotavirus infection occurred in week 9 of the study after the pigs were moved to a larger pen. The pigs were again given electrolyte therapy and all recovered well.

[0334] 3.2 Results This example showed that supplementation with L. rhamnosus FNZ129 did not adversely affect pig body weight (Table 11).

[0335] [Table 11]

[0336] Table 11 and Figure 1 show that supplementation with bacterial strain FNZ129 did not adversely affect weight gain in pigs.

[0337] 3.3 Conclusion Addition of L. rhamnosus FNZ129 to the diet had no significant negative effect on pig body weight. This study indicates that supplementation with FNZ129 can reduce methane emissions without adversely affecting weight gain.

[0338] 4. Example 4 - Effect of L. rhamnosus FNZ129 on the number of methane-producing bacteria in pigs 4.1 Materials and Methods 4.1.1 Animal studies: experimental design and animal ethics approval The piglets used in Example 3 were used in this study.

[0339] 4.1.2 Pig gastrointestinal sampling At 19 weeks of age, pigs were euthanized (capture bolt stunning, weighed, followed by exsanguination), the cecum and colorectal regions were tied off and removed, and digesta were collected from these two digesta regions. Digestive content samples were used for methanogen enumeration using the most probable number (MPN) method, and the remaining samples were used for volatile fatty acid (VFA) analysis by gas chromatography. For MPN analysis, 5 mL Eppendorfs were filled with cecal or colorectal contents for each animal and kept on ice until further processing in the laboratory. For VFA analysis, cecal contents were aliquoted into 50 mL Falcon tubes, whereas colorectal contents were sampled into 15 mL Falcon tubes and immediately kept on ice before storage at -20 °C.

[0340] 4.1.3 Most Probable Number (MPN) The MPN method (McCrady, 1918) was used to estimate the number of microorganisms capable of producing methane from samples of cecal and colorectal contents. Briefly, a measured amount of sample (approximately 1 g) was added to the first RM02 (Kenters et al. 2011) dilution tube, and the weight was recorded for use in the final calculation. From the first dilution tube, 10-fold serial dilutions (1 mL in 9 mL of RM02 medium) were made. Each dilution was mixed uniformly and then diluted a total of 10-fold under anaerobic conditions. A dilution was selected from the series, and the sample was inoculated into a Balch tube containing BRN-RF10 medium (Balch et al., 1979; Hoedt, 2017) supplemented with methanol (final concentration 100 mM) and pressurized with 180 kPa overpressure of H2+CO2. The inoculated BRN-RF10 medium tube was horizontally cultured at 39 °C for one month. These conditions allow the growth of not only methylotrophic methanogens, but also hydrogenotrophs such as methanogenic archaea and homoacetogens. After 1 month, the tubes were placed at room temperature for 30 min and then headspace gas analysis was performed using gas chromatography. Gas samples (0.5 mL) were collected from the tube headspace at the pressure inside the culture vessel using a polycarbonate 1 mL Luer-Lok syringe (Becton Dickinson and Co., Franklin Lakes, NJ, USA) fitted with a Mininert Luer-tip syringe valve (Hamilton, Reno, NV, USA). Headspace gas samples were manually injected into an Aerograph 660 gas chromatograph (Varian Associates, Palo Alto, CA, USA) equipped with a Porapak Q80 / 100 mesh column (Waters Corporation, Milford, MA, USA) and a thermal conductivity detector. N2 was used as the carrier gas. A 0.5 mL sample of a gas standard containing H2:CH4:N2 (5%:30%:65% v / v; BOC Gas, Palmerston North, NZ) was measured at 1 atm and used for calibration. The presence of methane was used as an indicator of methanogenic activity in the tube and was considered positive. If no methane was detected in the headspace, the tube was considered negative.By measuring the presence or absence of methane in the gas headspace of the culture tubes, it is possible to determine which tubes have methanogens present and metabolically active. From this data, and using the MPN table, the total number of methanogenic organisms present in the original sample was calculated.

[0341] 4.1.4 Medium preparation RM02 was prepared anaerobically and dispensed into Hungate tubes (9 mL per tube) under anaerobic conditions, then autoclaved at 125°C for 20 min. BRN-RF10 medium supplemented with 60 mM sodium formate (final concentration) was prepared and dispensed into Balch tubes under anaerobic conditions (9.8 mL per tube), then autoclaved at 125°C for 20 min. Prior to inoculation, 0.5% methanol (100 mM final) and 0.1 mL of Coenzyme M solution (10 μM) were added by syringe using anaerobic techniques. After injection, the tubes were pressurized with 180 kPa H2+CO2 overpressure (80:20, BOC Gases NZ).

[0342] 4.2 Results

[0343] [Table 12]

[0344] The results showed a significant reduction in methanogen counts and methanogenic activity in the cecum of pigs treated with FNZ129 (Table 12 and Figures 2 and 3). FNZ129 treatment also resulted in a numerical reduction in methanogen counts and methanogenic activity in colorectal samples, although the differences did not reach statistical significance.

[0345] 4.3 Discussion The porcine gut microbiota shares many similarities with humans, including the dominance of two major phyla, Bacteroidetes and Firmicutes, which occur in similar proportions in both species. Although their relative abundance varies, bacteria associated with human health, such as Lacticaseibacillus, Bifidobacterium, and Faecalibacterium, are also commonly found in pigs. These similarities in the microbiota are likely driven by similarities in the digestive system that lead to shared ecological constraints.

[0346] The MPN method allowed us to identify a significant effect of FNZ129 on cecal methanogens. Feeding the FNZ129 strain reduced the population levels of methanogens in the pig cecum, supporting the hypothesis that this strain is capable of producing biological compounds that inhibit the growth of methanogens.

[0347] Because methanogens in monogastric animals prefer the distal part of the digestive tract, we expected a lower methanogen population in the porcine ceca compared to the large intestine, regardless of treatment. The methanogen community estimated from 16S rRNA copy number was approximately 10 / g digesta from the ceca to the rectum. 8 ~10 9 Similarly, the proportion of methane in gas generated from the digestive tract ranges from 1.7–2.5% to 29–38% from the cecum to the rectum, respectively. There is an increasing gradient of methanogen population and methane formation in the porcine hindgut from the cecum to the rectum (Jorgensen et al. 2011, Gresse et al. 2019), reflecting slower transit times from the cecum to the colorectum, more anaerobic conditions, and higher pH. These conditions favor methanogens, making it easier for them to grow and maintain populations.

[0348] Probiotics administered to pigs first encounter methanogens after passing through the stomach and small intestine and entering the cecum. Since the number and activity of methanogens is low in the cecum, the effect of probiotic strains is likely to have the greatest anti-methanogen effect in this gut compartment. Furthermore, since the cecum represents a favorable niche for Lactobacilli, we also expect to see the effect of probiotics on microbial fermentation through changes in the VFA profile in this region of the gut. LAB are most active in this region and appear to have an inhibitory effect on the methanogen population, either by interacting with other members of the ecosystem or by producing and / or releasing compounds into the cecum.

[0349] 4.4 Conclusion This example shows that feed supplementation with L. rhamnosus FNZ129 can reduce the number of methanogenic microorganisms in the intestine of pigs.

[0350] 5. Example 5 - Effect of bacterial strains on volatile fatty acid production in pigs 5.1 Materials and Methods 5.1.1 Sample collection This experiment used the pigs used in Examples 3 and 4. Immediately after euthanasia, the pig cecum and colorectal region of the intestine were tied off and removed. The cecal contents were aliquoted into 50 mL Falcon tubes, while the colorectal contents were sampled into 15 mL Falcon tubes, immediately placed on ice, and transferred to a -20°C freezer until further analysis.

[0351] 5.1.2 Sample preparation for gas chromatographic analysis of VFAs Samples were thawed at room temperature. An aliquot (0.5 mL) of each sample was initially removed. The remaining sample volume was centrifuged at 21,000 × g for 10 min at 4 °C, and 0.9 mL of supernatant was removed and added to 0.1 mL of internal standard (20 mM 2-ethylbutyrate in 20% phosphoric acid), mixed, and frozen at -20 °C until analysis. After thawing and recentrifuging at 21,000 × g for 10 min at 4 °C, 0.2 milliliters of supernatant was taken and derivatized for non-VFA analysis by gas chromatography (Richardson et al. 1989). Meanwhile, the remaining samples were directly analyzed by gas chromatography (Attwood et al., 1998) using a gas chromatograph (Hewlett-Packard Model 6869, Montreal, MA, Canada) equipped with an autosampler, a Zebulon ZB-FFAP 30.0 m × 0.53 mm ID × 1 μm film column (Phenomenex, Torrance, CA, USA) and a flame ionization detector set at 265 °C.

[0352] 5.2 Results The main VFAs present in the cecal and colorectal samples were acetate, propionate, and butyrate, their respective concentrations and proportions corresponding to the normal levels of VFAs present in monogastric animals.

[0353] 5.2.1 Cecal VFA Cecal VFA concentrations in pigs fed FNZ129 were significantly different compared to control pigs. Cecal VFAs are reported as both absolute concentrations of acetate, propionate, and butyrate (Figure 4) and their respective relative proportions as a percentage of total VFAs (Table 13). Administration of the FNZ129 probiotic strain significantly reduced absolute acetate concentrations (Figure 4) and significantly increased the proportion of propionate to total VFAs (Table 13). Cecal concentrations of lactate and butyrate in pigs fed FNZ129 were not significantly different from the control group. No succinate was detected in any of the cecal contents, and total VFA concentrations were lower in FNZ129-treated animals (42.6 mM) compared to control animals (53.6 mM), although this difference was not significant.

[0354] [Table 13]

[0355] 5.2.2 Colon VFA Colorectal VFAs are also reported as both the absolute concentrations of acetate, propionate, and butyrate (Figure 5) and the relative percentage of each of these to total VFAs (Table 14). The results showed no effect on the colorectal fermentation process of L. rhamnosus FNZ129 (Table 14 and Figure 5). The percentage of acetate in the colon samples was higher than in the control, but the difference was not significant.

[0356] [Table 14]

[0357] 5.3 Discussion The MPN (Example 4) and VFA (Example 5) results show the effect of FNZ129 on the cecal fermentation process. Administration of FNZ129 reduced the absolute concentration of acetate in the cecum and increased propionate as a percentage of total VFA. The effect of the FNZ129 strain was only seen in the cecum, with no differences observed in the FNZ129 treatments in colorectal samples compared to the control group.

[0358] The change in propionate proportion may indicate an alteration in hydrogen metabolism in the cecum. A decrease in methanogenic activity in the cecum leads to an increase in hydrogen accumulation, while an increase in hydrogen concentration in the cecum leads to the thermodynamically favorable production of propionate. The observed differences in acetate and propionate in the cecum, combined with the lower methanogenic MPN results, suggest that the FNZ129 strain had an inhibitory effect on the cecal methanogenic community and / or altered the gastrointestinal microbiota, disrupting intermediate cross-feeding between members of the microbiota, potentially reducing their methane production.

[0359] 5.4 Conclusion This example shows that dietary supplementation with L. rhamnosus FNZ129 significantly increased the proportion of propionic acid, suggesting an alteration in hydrogen metabolism consistent with an inhibitory effect on cecal methanogens.

[0360] References

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[0368] [ka]

[0369] Industrial Applicability The present invention relates to the use of probiotic bacteria, in particular L. rhamnosus strain FNZ129 and / or derivatives thereof, in particular for improving the body weight or condition of animals, such as ruminants and / or monogastric animals; for improving feed efficiency, growth, productivity, and / or milk or meat yield; and / or for inhibiting the growth of methanogenic bacteria and / or archaea in the animal's digestive tract; for reducing the ability of the ruminal and / or gastrointestinal microflora to produce methane. Reduce methane emissions by animals, deliver microorganisms to animals, and / or reduce the greenhouse gas emission footprint of animals. Methods of using L. rhamnosus strain FNZ129 and / or derivatives thereof, as well as feed compositions comprising same, are also provided.

[0370] [ka]

[0371] [ka]

[0372] [ka]

Claims

1. Isolated Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

2. A food or feed composition comprising Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

3. 3. A food or feed composition according to claim 2, wherein the composition is a ruminant feed composition or a monogastric animal feed composition.

4. 1. A feed composition for improving the body weight and / or body composition of an animal, increasing feed efficiency in an animal, enhancing growth and / or productivity in an animal, increasing the yield of milk and / or milk components produced from the animal, inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal, reducing the methane-producing capacity of the gastrointestinal microflora, reducing methane emissions by the animal, delivering microorganisms to the animal, and / or reducing the greenhouse gas emission footprint of the animal, wherein the feed composition comprises Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof.

5. a. the feed composition is a ruminant feed composition and the animal is a ruminant; or 5. The feed composition of claim 4, wherein the feed composition is a monogastric feed composition and the animal is a monogastric animal.

6. 5. The feed composition of claim 4, wherein the feed composition is a fermented yogurt-style composition, the fermented yogurt-style composition being formed through a process of growing L. rhamnosus FNZ129 using a dairy-based carrier or a non-dairy-based carrier.

7. 5. The feed composition of claim 4, which is or comprises a partial or complete mixed ration (TMR), corn, soybeans, forage, grain, distillers grain, germinated grain, legume, fiber, forage, grass, hay, straw, silage, grain, leaf, meal, mash feed, fruit pulp, vegetable pulp, fruit or vegetable pomace, citrus meal, wheat shorts, corn cob meal, lick block, or molasses.

8. 5. The feed composition of claim 4, further comprising at least one microorganism of a different species or strain, a methanogen or a vaccine that inhibits methane production, and / or a natural or chemically synthesized methane production inhibitor, and / or a methane production inhibitor such as bromoform.

9. 5. The feed composition of claim 4, further comprising one or more agents selected from one or more prebiotics, one or more probiotics, one or more postbiotics, one or more dietary fiber sources, one or more galactooligosaccharides, one or more short-chain galactooligosaccharides, one or more long-chain galactooligosaccharides, one or more fructooligosaccharides, inulin, one or more galactans, one or more fructans, lactulose, or a mixture of any two or more thereof.

10. 5. The feed composition of claim 4, wherein the derivative of L. rhamnosus FNZ129 is a cell lysate of the strain, a cell suspension of the strain, a metabolic product of the strain, a culture supernatant of the strain, or killed L. rhamnosus FNZ129.

11. A ruminant feed composition as described in claim 4, wherein the composition comprises milk, milk powder, milk replacer, milk fortifier, colostrum, whey, whey powder, sucrose, maltodextrin, and / or rice hulls.

12. a. improving the body weight and / or body composition of an animal; b. Increases the feed efficiency of animals; c. enhancing animal growth and / or productivity; d. increasing the yield of milk and / or milk components produced by the animal; e. inhibiting the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of an animal; f. reducing the ability of the animal's gastrointestinal microflora to produce methane; g. Reducing methane emissions from animals; h. delivering microorganisms to animals; and / or i. A method for reducing greenhouse gas emissions from an animal, the method comprising: i. a food or feed composition according to claim 2 or 3, ii. The feed composition according to any one of claims 4 to 11, and / or iii. An effective amount of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof. to an animal.

13. 13. The method of claim 12, wherein the animal is a ruminant or a monogastric animal.

14. 13. The method according to claim 12, wherein the growth of methylotrophic methanogens, preferably methanogens from the genus Methanosphaera, is inhibited in the forestomach and / or caecum of an animal.

15. 13. The method of claim 12, wherein the L. rhamnosus FNZ129 or a derivative thereof is administered in a composition that is a food, beverage, food additive, beverage additive, animal feed, animal feed additive, animal feed supplement, dietary supplement, carrier, vitamin or mineral premix, nutritional product, enteral nutritional product, soluble, slurry, supplement, pharmaceutical, lick block, drench, tablet, capsule, pellet, bolus, or intraruminal product, or wherein the L. rhamnosus FNZ129 is encapsulated, for example, in a liposome, microbubble, microparticle, or microcapsule.

16. 16. The method of claim 15, wherein L. rhamnosus FNZ129 or a derivative thereof is administered in drinking water, milk, milk powder, milk replacer, milk fortifier, whey, whey powder, partial or complete mixed ration (TMR), feed pellets, corn, soybeans, feed, grains, distiller's grains, germinated grains, legumes, vitamins, amino acids, minerals, fiber, forage, grass, hay, straw, silage, grains, leaves, meal, solubles, slurry, supplements, mash feed, meal, fruit pulp, vegetable pulp, fruit or vegetable pomace, citrus meal, wheat shorts, corn cob meal, molasses, sucrose, maltodextrin, rice hulls, vermiculite, zeolite, or ground limestone.

17. To animals, a. 10 per kilogram of dry weight carrier feed ４ ~10 １３ colony forming units per kilogram of dry weight carrier feed, preferably 10 ８ ~10 １２ colony-forming units; b. 10 per kg of animal weight ４ ~10 １０ colony forming units / day, preferably 10 per kg of animal body weight ５ ~10 ８ colony-forming units / day; or c.10 ４ ~10 １３ colony forming units / day, preferably 10 ６ ~10 １３ 13. The method of claim 12, comprising administering L. rhamnosus FNZ129 in an amount of colony forming units / day.

18. 13. The method of claim 12, wherein the derivative of L. rhamnosus FNZ129 is a cell lysate of the strain, a cell suspension of the strain, a metabolic product of the strain, a culture supernatant of the strain, or killed L. rhamnosus FNZ129.

19. 13. The method of claim 12, further comprising administering at least one microorganism of a different species or strain, a methanogen or a vaccine that inhibits methane production, and / or a methane production inhibitor, such as a natural or chemically synthesized methane production inhibitor and / or bromoform.

20. 13. The method of claim 12, wherein L. rhamnosus FNZ129 or a derivative thereof is administered separately, simultaneously, or sequentially with one or more agents selected from one or more prebiotics, one or more probiotics, one or more postbiotics, one or more dietary fiber sources, one or more galactooligosaccharides, one or more short-chain galactooligosaccharides, one or more long-chain galactooligosaccharides, one or more fructooligosaccharides, inulin, one or more galactans, one or more fructans, lactulose, or a mixture of any two or more thereof.

21. The method is a. Increases animal growth or productivity; b. increasing the yield of milk and / or milk components produced by the animal; c. Increase the yield of milk fat, milk protein, or milk solids in the milk produced by the animal; and / or d) The method of claim 12, further improving the body weight and / or body composition of the animal.

22. 13. The method of claim 12, wherein the animal is a ruminant; preferably a cow, goat, sheep, bison, yak, buffalo, deer, camel, alpaca, llama, wildebeest, antelope, or nilgai; more preferably a cow or sheep; most preferably a cow.

23. 23. The method of claim 22, wherein the ruminant is a lactating animal.

24. 13. The method of claim 12, wherein the animal is a monogastric animal; preferably a human, pig, cat, dog, horse, donkey, rabbit, or poultry; more preferably a non-human animal; most preferably a pig, chicken, duck, goose, or turkey.

25. 25. The method of claim 24, wherein the monogastric animal is a companion animal.

26. 13. The method of claim 12, wherein the animal is a pre-weaned animal, such as a calf, lamb, piglet, or foal; or a post-weaned animal; or wherein L. rhamnosus FNZ129 is administered to the animal both pre-weaned and post-weaned.

27. 13. The method of claim 12, wherein the administration is to a pre-weaned animal and the inhibition of the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of the animal, the reduction in methane emissions by the animal, e.g., methane production, and / or the increase in feed efficiency in the animal persists after weaning.

28. 13. The method of claim 12, wherein the inhibition of the growth of methanogenic bacteria and / or archaea in the gastrointestinal tract of the animal, the reduction in methane emissions, e.g., methane production, by the animal, and / or the increase in feed efficiency in the animal persists for at least 2 days, 3 days, 5 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, or 7 years from the last administration of L. rhamnosus FNZ129; preferably for the lifetime of the animal.

29. An animal to which the method of any one of claims 12 has been applied.

30. 1. A method for producing an animal product, e.g., a dairy product, a meat product, or a wool product, with reduced greenhouse gas emissions, the method comprising: a. providing the animal of claim 29; and b. Producing animal products from animals The method includes:

31. Use of Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof to manufacture a composition for improving the body weight and / or body composition of an animal, increasing feed efficiency in an animal, increasing growth and / or productivity in an animal, increasing the yield of milk and / or milk components produced from the animal, inhibiting the growth of methane-producing bacteria and / or archaea in the gastrointestinal tract of an animal, reducing the ability of the gastrointestinal microflora to produce methane, reducing methane emissions by an animal, delivering microorganisms to an animal, and / or reducing the greenhouse gas emission footprint of an animal.

32. 32. The use according to claim 31, wherein the composition comprises a food or feed composition according to claim 2 or 3, or a feed composition according to any one of claims 4 to 11.

33. Lacticaseibacillus rhamnosus (Lactobacillus rhamnosus) strain FNZ129 (NMIA accession number V21 / 015446 dated August 2, 2021) or a derivative thereof for use in improving the body weight and / or body composition of an animal, increasing the feed efficiency of an animal, enhancing the growth and / or productivity of an animal, increasing the yield of milk and / or milk components produced from an animal, inhibiting the growth of methane-producing bacteria and / or archaea in the gastrointestinal tract of an animal, reducing the ability of the gastrointestinal microflora to produce methane, reducing methane emissions by an animal, delivering microorganisms to an animal, and / or reducing the greenhouse gas emission footprint of an animal.

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