Use of lignin fractions as supplement ingredients for human and animal foods

Abstract

The use of the lignin fraction as a food supplement ingredient for humans and animals, and food supplements containing same, are disclosed.

Description

[Technical Field]

[0001] The present invention relates to the use of a lignin fraction as a food supplement ingredient for humans and animals, and to food supplements containing same. [Background technology]

[0002] In some industries, for example in the food industry, it is necessary to avoid bacterial growth in order to maintain the hygiene of the products processed in the industry, for example food. On farms and in slaughterhouses, bacterial growth is on the one hand very common and on the other hand very important to limit and avoid its growth.

[0003] Known products are often ineffective against many microorganisms, which necessitates the use of many different products, and they often contaminate both the environment and the products in which they are used.

[0004] In such cases, the products typically used are antibiotics, but the overuse and misuse of these drugs, along with a lack of new drug development by the pharmaceutical industry due to declining economic incentives and stringent regulatory requirements, are thought to underlie the antibiotic resistance crisis.

[0005] Antimicrobial resistance (AMR) is the ability of microorganisms to resist the effects of drugs previously used in treatment. The term also includes the more specific term "antibiotic resistance." It applies only to bacteria that have become resistant to antibiotics. Resistant microorganisms are more difficult to treat and require alternative drugs or higher doses, both of which may be more expensive or more toxic.

[0006] The World Health Organization (WHO) has asserted that the inappropriate use of antibiotics in livestock farming is a fundamental cause of the emergence and spread of antibiotic-resistant bacteria, and that the use of antibiotics as growth promoters in animal feed should be restricted. The World Organization for Animal Health (WHO) has added a set of guidelines to its Terrestrial Animal Health Code recommending to member states on the development and harmonization of national antimicrobial resistance surveillance and monitoring programs, the monitoring of antibiotic use in livestock farming, and recommendations for ensuring the appropriate and prudent use of antibiotics. Another set of guidelines is for establishing relevant risk factors and implementing methodologies that can help assess the risk of antibiotic resistance.

[0007] Thus, there is a felt need to effectively combat these harmful microorganisms while avoiding the use of antibiotics and at the same time maintaining human and animal health. Summary of the Invention

[0008] The above object has been achieved by the use of a lignin fraction as an antipathogenic substance in a food supplement for humans and animals, as claimed in claim 1.

[0009] In this respect, the present invention also relates to a food supplement comprising said lignin fraction and a suitable food carrier.

[0010] In another aspect, the present invention relates to a food supplement for use in feeding livestock, said food supplement being administered in an amount effective to improve the quality and shelf life of meat obtained from the livestock.

[0011] In a further aspect, the present invention relates to a food supplement for use in feeding female livestock, said food supplement being administered in an amount effective to improve the yield and quality of milk obtained from the female livestock.

[0012] In a further aspect, the present invention relates to a food supplement for use in feeding livestock, said food supplement being administered in an amount effective to improve the reproductive performance of the livestock.

[0013] In a further aspect, the present invention relates to a food supplement for use in feeding livestock, said food supplement being administered in an amount effective to reduce methane emissions without impairing digestive efficiency or rumination.

[0014] The features and advantages of the present invention will become apparent from the following detailed description, the examples provided for illustrative purposes, and the accompanying drawings. [Brief explanation of the drawings]

[0015] [Figure 1] FIG. 1 shows the redness profile of meat during 14 days of aging for the control and experimental groups according to Example 6. [Figure 2] FIG. 2 shows the yellowness profile of meat during 14 days of aging for the control and experimental groups according to Example 6. [Figure 3] FIG. 3 shows total gas production during 24 hours of in vitro digestion of ruminal fluid according to Example 6. DETAILED DESCRIPTION OF THE INVENTION

[0016] The subject of the present invention is therefore the use of a lignin fraction as an antipathogenic substance in human and animal food supplements in the prevention and treatment of infectious diseases caused by food pathogens, said lignin fraction comprising fragments with a weight-average molecular weight of up to 2,500 daltons as determined by size exclusion chromatography, said fragments comprising an average of up to 13 phenylpropane units by weight.

[0017] Lignin is a complex organic polymer that forms the important structural material of the support tissues of some algae, vascular plants (including their bark), and herbaceous plants, such as trees (i.e., conifers and broad-leaved trees), all cereal straw, sugarcane bagasse, grasses, hemp, jute, hemp, or cotton. Lignin may also have mineral origins, such as peat, leonardite, or coal. Chemically, lignin is a highly irregular, randomly cross-linked polymer of phenylpropane units with a weight-average molecular weight of more than 20,000 daltons, linked by many different bonds. A representative and exemplary lignin fragment (I) containing the most important bond patterns is shown below:

[0018] [ka]

[0019] The polymer is the result of enzyme-mediated dehydrogenative polymerization of three phenylpropanoid monomer precursors:

[0020] [ka]

[0021] Each part gives:

[0022] [ka]

[0023] Coniferyl alcohol is present in all species and is the major monomer in conifers. Deciduous tree species contain up to 40% sinapyl alcohol units, while grasses and crops may also contain coumaryl alcohol units.

[0024] Lignin is classified into softwood lignin and hardwood lignin depending on the biomass source from which it is derived. Suitable starting biomass sources for obtaining suitable lignin fractions include any lignin, including substantially pure lignin, kraft lignin, biomass-derived lignin, lignin from alkaline pulping processes, lignin from soda processes, lignin from organosolv pulping, and any combination thereof.

[0025] The expression "substantially pure lignin" is to be understood as at least 90% pure lignin, preferably at least 95% pure lignin, based on the dry raw biomass, the remainder being extractives, carbohydrates such as hemicellulose, and inorganic matter.

[0026] The expression "kraft lignin" is understood to mean lignin derived from kraft black liquor. Black liquor is an alkaline aqueous solution of lignin residues, hemicellulose, and inorganic chemicals used in the kraft pulping process. Black liquor from the pulping process contains components derived from various softwood and hardwood species in varying proportions. Lignin can be separated from black liquor by various methods, including precipitation and filtration. Lignin typically begins to precipitate at pH values ​​below 11-12. Different pH values ​​can be used to precipitate lignin fractions with different properties. These lignin fractions are characterized by their molecular weight distribution (e.g., M w and M n The precipitated lignin may differ from one another by its size, polydispersity, hemicellulose and extractive content, and inorganic content. The precipitated lignin can be purified from inorganic impurities, hemicellulose, and wood extractives using an acid wash process. Further purification can be achieved by filtration.

[0027] Alternatively, lignin can be separated from pure biomass. The separation process can begin with liquefying the biomass with strong alkali, followed by a neutralization process. After alkali treatment, lignin can be precipitated in a manner similar to that presented above.

[0028] Alternatively, the separation of lignin from biomass involves an enzymatic treatment step, which alters the lignin extracted from the biomass. The lignin separated from the pure biomass is substantially sulfur-free (less than 3% sulfur content) and therefore amenable to further processing.

[0029] Preferably, the lignin so separated is also subjected to a depolymerization step to further reduce the weight average molecular weight of the fragments.

[0030] Preferably, the lignin so separated is also subjected to a depolymerization step to further reduce the weight and number average molecular weight of the fragments.

[0031] Suitable depolymerization processes include base-catalyzed depolymerization, acid-catalyzed depolymerization, metal-catalyzed depolymerization, ionic liquids-assisted depolymerization, and supercritical fluids-assisted lignin depolymerization.

[0032] In a preferred embodiment, said lignin fraction is obtained by base-catalyzed depolymerization.

[0033] Preferably, said lignin fraction is obtained by subjecting the separated lignin to base-catalyzed depolymerization at a temperature below 300°C and a pressure below 30 MPa.

[0034] The pH is set between 11 and 14 by adding a base such as NaOH, KOH, Ca(OH)2, LiOH, K2CO3, or mixtures thereof.

[0035] For the purposes of the present invention, the weight average molecular weight (M wMolecular weights of the components of a polymer sample must be calibrated using standard polymers of known weights. Values ​​from unknown samples are then compared to a calibration graph. Retention times depend on the column material, eluent, and similarity of the standard compared to the sample. In the present invention, the eluent is preferably 0.1 M NaOH.

[0036] The lignin fraction of the present invention has unexpectedly and surprisingly proven to be highly selective and effective against food pathogens without adversely affecting probiotic microorganisms, as shown in the examples provided below.

[0037] Food pathogens are gram-positive and gram-negative bacteria and fungi such as Escherichia coli, Candida albicans, Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella enteritidis, Campylobacter jejuni, and Listeria monocytogenes.

[0038] Examples of probiotic microorganisms include Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Enterococcus faecium, Saccaromyces boulardii, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium lactis, and Lactobacillus reuteri. reuteri, Lactobacillus brevis, Lactobacillus fermentum, Lactobacillus paracasei, Streptococcus thermophilus, etc.

[0039] This means that the lignin fractions of the present invention advantageously allow for the replacement of antibiotic treatment. Antibiotic resistance is defined as the emergence (and spread) of factors of bacterial resistance to antibiotics and is caused by the selective pressure exerted on microbial populations by the excessive and / or inappropriate use of antibiotics. The lignin fractions described herein have proven to be effective alternatives to antibiotics for the prevention and treatment of antibiotic-resistant infections.

[0040] As can be seen from the examples provided below, the lignin fraction of the present invention is advantageously and surprisingly capable of improving the overall health and wellness of humans and animals, particularly livestock, while improving the economic performance of the latter and the quality of the milk and meat obtained therefrom.

[0041] Preferably, the lignin fraction comprises fragments having a weight average molecular weight of up to 2,000 Daltons.

[0042] In a preferred embodiment, the lignin fraction comprises fragments having a weight average molecular weight of up to 1,500 Daltons.

[0043] In some embodiments, the lignin fraction comprises fragments having a weight average molecular weight down to 150 Daltons.

[0044] In a preferred embodiment, the lignin fraction comprises fragments having a weight average molecular weight of 150 Daltons to 2,500 Daltons, preferably fragments having a weight average molecular weight of 250 Daltons to 2,000 Daltons, more preferably fragments having a weight average molecular weight of 500 Daltons to 1,800 Daltons.

[0045] Preferably, in these embodiments, the fragments contain, on average by weight, up to 12 phenylpropane units, and more preferably, on average by weight, up to 11 phenylpropane units.

[0046] The molecular weights of the three phenylpropanoid monomer precursors vary between 150 Da for coumaryl alcohol, 180 Da for coniferyl alcohol, and 210 Da for sinapyl alcohol. Therefore, the average weight was 180 Da, and this value was used as the "phenylpropane unit." M w The value was divided by 180 Da to give the weight average number of phenylpropane units.

[0047] In a particularly preferred embodiment, the lignin fraction comprises fragments having a weight average molecular weight of 250 daltons to 2,000 daltons and a weight average of 2 to 11 phenylpropane units.

[0048] In another embodiment, the lignin fraction has a number average molecular weight (M n )

[0049] For the purposes of the present invention, the number average molecular weight (M n ) is measured by size exclusion chromatography. Preferably, the lignin fraction has a number average molecular weight (M) of up to 1,500 Daltons. n )

[0050] In a preferred embodiment, the lignin fraction comprises fragments having a number average molecular weight of between 150 and 1,000 daltons.

[0051] Without wishing to be bound by any theory, it is believed that the lower the number average molecular weight, the more active molecules there are. This is proposed considering that lower molecular weight means smaller fragments, smaller fragments mean fewer cross-linked / short chain fragments, and fewer cross-linked / short chain fragments means more free functional groups on them, meaning more fragments that are more reactive.

[0052] Furthermore, smaller molecules may be able to more easily penetrate and diffuse through the pathogen cell membrane, significantly improving the overall efficacy of the lignin fraction.

[0053] Preferably, in these embodiments, the fragments contain a number average of up to 11 phenylpropane units, more preferably a number average of up to 8 phenylpropane units.

[0054] The molecular weights of the three phenylpropanoid monomer precursors vary between 150 Da for coumaryl alcohol, 180 Da for coniferyl alcohol, and 210 Da for sinapyl alcohol. Therefore, the average weight was 180 Da, and this value was used as the "phenylpropane unit." M n The value was divided by 180 Da to give the number average number of phenylpropane units.

[0055] In a preferred embodiment, the lignin fraction has a weight average molecular weight (M) of 150 Daltons to 2,500 Daltons. w ) and fragments with number average molecular weights (M) up to 2,000 Daltons. n )

[0056] More preferably, the lignin fraction has a weight average molecular weight (M w ) of 150 Daltons to 2,500 Daltons, with a weight average of 2 to 13 phenylpropane units, and n ) contains fragments with a maximum size of 2,000 daltons and a number average of a maximum of 11 phenylpropane units.

[0057] In a further embodiment, the lignin fraction has a polydispersity index (PDI) of 1.25 to 6.

[0058] Polydispersity index (PDI) or heterogeneity index, or simply dispersity, is the degree of distribution of molecular weights in a particular polymer sample. PDI is the weight average molecular weight (M w ) to the number average molecular weight (M n ) which gives the distribution of individual molecular weights within a batch of polymer.

[0059] In a particularly preferred embodiment, the lignin fraction has a weight average molecular weight (M w ) and fragments having a weight average of 2 to 13 phenylpropane units, the lignin fraction having a polydispersity index of 1.25 to 6.

[0060] In a particularly preferred embodiment, the lignin fraction has a number average molecular weight (M n ) and fragments with a number average of up to 11 phenylpropane units, the lignin fraction having a polydispersity index of 1.25 to 6.

[0061] In a most preferred embodiment, the lignin fraction has a weight average molecular weight (M w ) and a weight average of 2 to 13 phenylpropane units, with a number average molecular weight (M n ) and fragments having a number average of up to 11 phenylpropane units, the lignin fraction having a polydispersity index of 1.25 to 6.

[0062] In a particularly preferred embodiment of the present invention, the lignin fraction has a weight average molecular weight (M w ), weight average of 4 to 8 phenylpropane units, number average molecular weight (M) of 300 to 700 daltons n ), and includes fragments having a number average of 2 to 4 phenylpropane units. In these particularly preferred embodiments, the most abundant phenylpropane units in the lignin fraction are derived from coniferyl alcohol, while the less abundant phenylpropane units are derived from sinapyl alcohol.

[0063] The lignin fraction may be in solid or liquid form.

[0064] When the lignin fraction is in a solid form, said solid form may be a tablet, mini-tablet, micro-tablet, granule, micro-granule, pellet, multi-particulate, micronized particle or powder.

[0065] When the lignin fraction is in liquid form, said liquid form is a solvent solution.

[0066] Suitable solvents are water, glycols, alcohols, polyhydric alcohols, organic acids, and combinations thereof.

[0067] Preferred solvents are water, methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, allyl alcohol, 1,2-propylene glycol, 1,3-propylene glycol, 1,2-ethylene glycol, polyethylene glycol (PEG), glycerol, lactic acid, polylactic acid, and mixtures thereof.

[0068] More preferred solvents are water, 1,2-propylene glycol, 1,3-propylene glycol, 1,2-ethylene glycol, polyethylene glycol (PEG), and mixtures thereof.

[0069] In the most preferred embodiment, the solvent is water.

[0070] Preferably, when the lignin fraction is in a liquid form, the liquid form has a pH of 8 to 11, more preferably 9.5 to 10.5.

[0071] In a preferred embodiment, the lignin fraction is used in an amount of up to 10 kg per tonne of animal feed, more preferably 1 to 5 kg per tonne of animal feed.

[0072] In a further aspect, the present invention also relates to a food supplement comprising the lignin fraction for use as described above, and a suitable food carrier. The food supplement may be in solid or liquid form.

[0073] When the food supplement is in a solid form, said solid form can be a tablet, mini-tablet, micro-tablet, granule, micro-granule, pellet, multi-particulate, micronized particle or powder.

[0074] When the food supplement is in solid form, said solid form comprises up to 99 wt% of the lignin fraction, preferably 5-90 wt% of the lignin fraction.

[0075] When the food supplement is in liquid form, the liquid form can be a solution, emulsion, dispersion, suspension, gel, drop or spray.

[0076] When the food supplement is in liquid form, the liquid form contains up to 50 wt% of the lignin fraction, preferably 0.1-25 wt% of the lignin fraction, meaning that the composition is a concentrate that can be appropriately diluted with water as needed before use, or mixed directly with animal feed.

[0077] Suitable carriers include acidifying agents, acid neutralizing agents, anti-agglomerating agents, antioxidants, fillers, resist agents, gelling agents, coating agents, modified starches, sequestering agents, thickeners, sweeteners, thinners, solvents, dissociating agents, glidants, dyes, binders, lubricants, stabilizers, adsorbents, preservatives, humectants, flavorings, film-forming substances, emulsifiers, wetting agents, release agents, and mixtures thereof.

[0078] In a preferred embodiment, the food supplement further comprises a lignin fraction containing fragments having a weight-average molecular weight of 3,500 to 5,500 daltons as measured by size exclusion chromatography, the fragments containing an average of up to 40 phenylpropane units by weight. Preferably, the food supplement further comprises a lignin fraction containing fragments having a weight-average molecular weight of 4,000 to 5,000 daltons as measured by size exclusion chromatography, the fragments containing an average of up to 35 phenylpropane units by weight.

[0079] In another preferred embodiment, the food supplement further comprises at least one resin acid, preferably abietic acid, dehydroabietic acid, palustric acid, neoabietic acid, pimaric acid, isopimaric acid, sandalopimaric acid, or an ester thereof, or an ether thereof, or an alkali or alkaline earth salt thereof, or a mixture thereof.

[0080] Preferably, the food supplement contains said at least one resin acid in an amount of up to 10% by weight, more preferably up to 7% by weight, based on the weight of the food supplement.

[0081] Resin acids are found in conifers, and there are three main types of resin acid products: tall oil rosin (TOR), wood rosin, and gum rosin. TOR is the resin acid fraction isolated by vacuum distillation from crude tall oil (CTO) produced during pulp production. CTO is obtained by acidifying crude tall oil soap or crude sulfate soap (TOS). TOS is isolated from the cooking liquid in the pulp mill, often referred to as black liquor, during the pulp manufacturing process. Wood rosin is a fraction separated by steam distillation or other means from dead wood, tree stumps, branches, etc., while gum rosin is a resin fraction separated by steam distillation or other means from resin harvested from living trees, often referred to as taps.

[0082] Resin acid-containing materials obtained by vacuum distillation of crude tall oil include distilled tall oil (DTO), tall oil fatty acids (TOFA), and tall oil pitch (TOP). DTO contains 10-40% resin acids. CTO typically contains 15-70% resin acids, with the lowest resin acid content generally provided by cooking mixed wood pulp.

[0083] The term "tall oil rosin" or "TOR" should be understood to refer to a composition obtained by distillation of crude tall oil and further refining of the distilled tall oil. TOR typically contains 60-99% (w / w) resin acids.

[0084] The term "wood rosin" should be understood to refer to a composition obtained by distillation or other means from dead trees, tree stumps, branches, etc. Wood rosin typically contains 50-99% (w / w) resin acids.

[0085] The term "gum rosin" should be understood to refer to a composition obtained by distillation or other means of separation from resin harvested from living trees. Gum rosin typically contains 50-99% (w / w) resin acids.

[0086] The term "distilled tall oil" or "DTO" should be understood to refer to a composition obtained by distillation of crude tall oil and further refining of the distilled tall oil. DTO typically contains 10-60% (w / w) resin acids.

[0087] Resin acid-based compositions TOR, wood rosin, gum rosin, CTO, TOS and DTO can also be prepared by mixing one or more resin acid compositions with one or more fatty acid compositions in the form of an oil or fat, and the resin acid derivatives produced are, for example, esters, ethers or alkali metal salts.

[0088] Resin acids are known to exhibit many properties, such as antibacterial, anti-inflammatory, antioxidant, antibacterial biofilm properties, etc. However, resin acids have poor stability over time, especially in solid form, due to exposure to oxidation, spontaneous combustion, and packing.

[0089] It has surprisingly been found that when the at least one resin acid is mixed with the lignin fraction of the present invention, the resulting mixture, either solid or liquid, is very stable over time and does not oxidize, making it possible to fully benefit from the properties of the resin acids. This means that in the resulting food supplement containing the lignin fraction of the present invention, not only are the resin acids protected from degradation, but a synergistic anti-inflammatory effect between the lignin fraction and the resin acids is observed.

[0090] Food supplements are also available. 12 ~C 24 It may further comprise at least one alkali or alkaline earth salt of a fatty acid, at least one fatty oil, or a mixture thereof, which allows for better solubilization of the resin acids when a liquid form of the dietary supplement is preferred.

[0091] Preferably, the alkali or alkaline earth salt is a lithium, sodium, potassium, magnesium, calcium salt or mixtures thereof.

[0092] Preferably, the C 12 ~C 24The fatty acids are lauric acid (C12), tridecylic acid (C13), myristic acid (C14), pentadecylic acid (C15), palmitic acid (C16), margaric acid (C17), stearic acid (C18), oleic acid (C18:1), linoleic acid (C18:2), α-linolenic acid (C18:3), γ-linolenic acid (C18:3), nonadecylic acid (C19), arachidic acid (C20), heneicosanoic acid (C21), behenic acid (C22), tricosyl acid (C23), lignoceric acid (C24), and sucralose. Thearidonic acid (C18:4), eicosapentaenoic acid (C20:5), docosahexaenoic acid (C22:6), dihomo-γ-linolenic acid (C20:3), arachidonic acid (C20:4), adrenic acid (C22:4), palmitoleic acid (C16:1), vaccenic acid (C18:1), paulic acid (C20:1), elaidic acid (C trans-18:1), gondoic acid (C20:1), erucic acid (C22:1), nervonic acid (C24:1), mead acid (C20:3), or mixtures thereof.

[0093] In a preferred embodiment, the C 12 ~C 24 The at least one alkali or alkaline earth salt of a fatty acid is calcium palmitate, calcium laurate, calcium oleate, calcium soap of palm oil, or mixtures thereof.

[0094] Preferably, the at least one fatty oil is hemp oil, canola oil, sunflower oil, olive oil, corn oil, palm oil, coconut oil, pine oil, cottonseed oil, wheat germ oil, soy oil, safflower oil, linseed oil, tung oil, castor oil, soybean oil, peanut oil, rapeseed oil, sesame seed oil, rice germ oil, fish oil, whale oil, marine oil or mixtures thereof.

[0095] Preferably, the food supplement comprises the C 12 ~C 24 The at least one alkali or alkaline earth salt of a fatty acid, at least one fatty oil, or a mixture thereof is contained in a concentration of 1 to 100 kg per ton of the food supplement.

[0096] In another aspect, the present invention relates to a food supplement for use in feeding livestock to improve the quality and shelf life of meat obtained from livestock, in particular the present invention relates to a food supplement for use in feeding livestock, wherein said food supplement is administered in an amount effective to improve the quality and shelf life of meat obtained from livestock.

[0097] The term "livestock" in the present invention is intended to include both ruminant and non-ruminant animals. Ruminant animals are herbivorous mammals with a four-chambered stomach, in which they ferment and digest otherwise indigestible plant material, ruminating it (coughing up a ball of semi-digested plant material, chewing it again, and swallowing it again). Ruminant animals include cattle, sheep, goats, deer, antelopes, giraffes, and camels, as well as related animals such as bison, musk oxen, okapi, and llamas. Non-ruminant animals, including pseudoruminants, include pigs, horses, chickens, rabbits, turkeys, ducks, geese, quails, pheasants, partridges, and fish. This means that the food supplement of the present invention can advantageously be used in feeding fish and shellfish, i.e., in aquaculture.

[0098] Surprisingly, it has been observed that livestock fed the food supplement of the present invention provide superior quality meat, as the natural tendency of meat to lose its reddish or yellowish color over time is unexpectedly and significantly reduced, while at the same time the shelf life and freshness of the meat is dramatically increased (i.e., up to 7 days or more). Without wishing to be bound by any theory, it is believed that the highly effective antioxidant properties of the lignin fraction of the present invention, in addition to the antipathogenic properties reported above, inhibit the oxidation process on the surface of the meat, thereby counteracting color changes and allowing the meat to remain fresh and edible for longer periods.

[0099] Color is an important factor commonly used as a quality indicator in the meat industry and meat science research. Because consumers use discoloration as an indicator of freshness and a lack of wholesomeness, color is reported to be one of the most important meat quality attributes and significantly influence purchasing decisions. Consumer-perceived quality does not always correspond to objective quality, but either way, it can result in significant economic losses in the fresh meat market. Color is also important from an economic perspective, as it signals quality to consumers who prefer to purchase red meat rather than brown, and the industry incurs losses due to undesirable color. As demonstrated in the following examples, the food supplement of the present invention enables fresh meat to maintain highly satisfactory levels of redness and yellowness over an extended period of time.

[0100] In a further aspect, the present invention relates to a food supplement for use in feeding female livestock to improve the production and quality of milk obtained from the female livestock, in particular, the present invention relates to a food supplement for use in feeding female livestock, wherein said food supplement is administered in an amount effective to improve the production and quality of milk obtained from the female livestock.

[0101] Surprisingly, it has been observed that female livestock fed the food supplement of the present invention provide greater amounts of milk per day and of better quality, as shown in the examples below. Indeed, the effect of administering a food supplement to lactating dairy cows is evident in several events and timelines, based on the following observations regarding the quantity and quality of milk produced: - Increased concentration of total polyphenols in milk; - Increased daily milk production per animal; - Decreased somatic cell count in the acute phase; - Increased absorption capacity of free radicals (TEAC).

[0102] The above results are highly significant with a peak of approximately 250% in total polyphenol concentration, an increase in overall milk production of +20.8%, a decrease in somatic cell count of -31%, and an increase in TEAC of +33%.

[0103] It should also be noted that these results show a particular synchronization, with the maximum concentration occurring around days 15-17 of treatment.

[0104] It was also found that supplementation according to the present invention produced a condition that extended the benefits for at least two weeks after cessation. The beneficial effects began to wane after about 20 days, as evidenced by a regrowth of somatic cells in the milk.

[0105] Since cow's milk usually contains only trace amounts of polyphenols, it is highly appreciated that milk obtained from cows fed the food supplement of the present invention unexpectedly contains up to 460 mg / l of polyphenols. Considering that the human requirement for polyphenols is 820 mg / die and that the average human intake of milk is 250 ml / die, the contribution of polyphenols provided by the milk of the present invention is 114 mg, when as stated, this is usually close to zero.

[0106] In a further aspect, the present invention relates to a food supplement for use in feeding livestock to improve livestock fertility, in particular, the present invention relates to a food supplement for use in feeding livestock, wherein said food supplement is administered in an amount effective to improve livestock fertility.

[0107] Surprisingly, livestock fed the food supplement of the present invention have been shown to have significantly increased birth rates for both male and female livestock.

[0108] In particular, female livestock such as dairy cows will see an increase in fertility in terms of conception rate after insemination (at least +15%) and reproductive lifespan, while male livestock such as bulls and buffaloes will see an increase in fertility in terms of semen quality, reproductive parameters and reproductive lifespan.

[0109] In a further aspect, the present invention relates to a food supplement for use in feeding livestock to reduce methane emissions without impairing digestive efficiency or rumination, in particular the present invention relates to a food supplement for use in feeding livestock, wherein said food supplement is administered in an amount effective to reduce methane emissions without impairing digestive efficiency or rumination.

[0110] Emissions of the greenhouse gas methane from livestock are higher than previously thought, posing an additional challenge in the fight to limit global warming. Revised calculations of methane production per cattle indicate that global livestock emissions in 2011 were 11% higher than estimates based on data from the United Nations Intergovernmental Panel on Climate Change (IPCC). In addition to natural sources such as peat bogs, wetlands, and termites, human methane (accounting for about two-thirds of the total) is produced in two ways: through the escape of an odorless, colorless gas during the production and transportation of coal, oil, and especially natural gas; and, to a similar extent, through the burping and belching of ruminant animals such as cattle and sheep, and the decay of organic waste, especially in landfills. According to the IPCC, methane accounted for approximately 16% of global greenhouse gas emissions in 2015. Methane is a much more potent greenhouse gas than CO2, capturing more of the sun's radiative power, but its atmospheric persistence is shorter. Taking that into account, scientists calculate that the gas's "global warming potential" is 28 times greater than that of carbon dioxide over a 100-year period. Methane emissions from livestock are increasing most rapidly in rapidly developing regions of Asia, Latin America, and Africa. In contrast, growth has slowed sharply in the United States and Canada.

[0111] Therefore, the potential to reduce overall methane production from ruminant livestock was highly appreciated, and at the same time, the fact that the lignin fraction of the present invention can achieve this objective without compromising digestive efficiency or rumination, as shown in the examples below, was highly unexpected.

[0112] Preferably, the food supplement is administered to ruminants in a daily dose containing up to 100 g of the lignin fraction of the present invention, more preferably 20-80 g of the lignin fraction of the present invention.

[0113] It should also be understood that all combinations of the preferred embodiments of the use of the lignin fraction of the present invention and the use of food supplements, and the uses thereof reported above, are considered to be disclosed herein.

[0114] It is to be understood that all combinations of the uses of the lignin fractions of the present invention, food supplements, and preferred aspects of each use disclosed above are described herein.

[0115] The following are examples of the present invention provided for illustrative purposes. [Example]

[0116] In this embodiment, M w and M n was measured by size exclusion chromatography according to the following procedure.

[0117] Reagents and materials - Eluent: 0.1M NaOH; flow rate 0.5ml / min - RI detector calibration: pullulan standard; M p :100,000~1,080(6 standard)(M p is the peak maximum molecular weight) - Calibration of UV detector (280 nm): PSS standard; polystyrene sulfonate sodium salt; M p 65,400~891 (6 standards); Dissolve the standard substance in ultrapure water to a concentration of approximately 5mg / ml. Injection volume is 20μl. -Quality control sample: M w Lignin with a known distribution is used.

[0118] Facilities and Equipment - Dionex Ultimate 3000 Autosampler, Column Compartment and Pump - Dionex Ultimate 3000 Diode Array Detector - Refractive index detector: Shodex RI-101 - Columns: PSS MCX column: pre-column and two analytical columns: 1000 Å and 100,000 Å; the column material is a sulfonated divinylbenzene copolymer matrix. - Syringe filter 0.45μm and glass sample bottle for STD samples. Sample filtration: Mini-Uniprep syringeless filter device PTFE or nylon; 0.45 μm. Optionally, a 5 μm syringe filter for pre-filtration. - Measuring bottle

[0119] procedure - Preparation of eluent Ideally, the water used to prepare the eluent is high-quality deionized water with low resistivity (18 MΩ cm or better) that is as free as possible from dissolved carbon dioxide. The water should be free of biological contaminants (e.g., bacteria and mold) and particulate matter. Ideally, the water used to prepare the eluent is high-quality deionized water with low resistivity (18 MΩ cm or better) that is as free as possible from dissolved carbon dioxide. The water should be free of biological contaminants (e.g., bacteria and mold) and particulate matter. - Needle wash with 10% MeOH / water

[0120] - Liquid samples The strong alkaline solution sample is diluted 1:100 and filtered through a PTFE syringe filter (0.45 μm) into a vial. The solid lignin sample is diluted and dissolved in 0.1 M NaOH and filtered through a PTFE syringe filter (0.45 μm). The prepared sample is loaded into the autosampler. The injection volume is 20 μl. After the sample, 1 M NaOH is injected as a sample to wash the column.

[0121] Instrument parameters - Flow rate 0.5 ml / min - Eluent 0.1M NaOH - Column oven temperature 30℃ - Isocratic elution - Runtime 48 minutes

[0122] - Solid samples Dry the solid sample (lignin) overnight in a 60°C oven if necessary. Weigh approximately 10 mg into a 10 ml measuring bottle. Dissolve and dilute the sample in 0.1 M NaOH solution and fill to the mark. Filter the sample through a PTFE, 0.45 μm filter. If the sample does not dissolve properly, it can be placed in an ultrasonic water bath or filtered through a 5 μm syringe filter.

[0123] - Calibration standards Approximately 50 mg of each standard is weighed into a 10 ml measuring bottle and filled to the mark with ultrapure water. The standards are filtered through a PTFE, 0.45 μm syringe filter. After running the calibration samples, the calibration results are integrated into the processing method, processed, and saved. The calibration is a linear, first-order calibration.

[0124] - Quality control samples Lignin samples were used as quality control samples. w Lignin with a known distribution is used. The lignin is dissolved in 0.1 M NaOH to a concentration of approximately 1 mg / ml.

[0125] Example 1 Organosolv lignin obtained from beech wood (Fagus sylvatica) was treated with base-catalyzed depolymerization (BCD). The BCD process was carried out at 280 °C, 250 bar, and pH 12-14 for 8 minutes. The resulting lignin product consisted of a liquid fraction and a solid fraction. These fractions were then separated.

[0126] The liquid lignin fraction was an oil and had the following properties: Single species: Fagus sylvatica M w : 100~300 Da (1~2 phenylpropane units) Phenol: 0% Guaiacol: 15-20% Syringol: 50-60% Catechol and methoxycatechol: 5-10% Oligomer / Unknown: 15-30%

[0127] The solid lignin fraction had the following properties: Single species: Fagus sylvatica M w : 800-1,500 Da (4-8 phenylpropane units) M n : 300-700 Da (2-4 phenylpropane units) OH group structure: Aliphatic 0.2~0.4mmol / g Carboxylic acid 0.3~0.5mmol / g Condensed and syringyl 1.0-2.0mmol / g Guaiacil 0.4mmol / g Catechol and p-OH-phenyl 1.0-1.8mmol / g

[0128] Example 1a. 50 g of the above oily lignin fraction (5% w / w) was mixed with 950 g of 1,3-propylene glycol and heated at 40 to 50°C. The mixture was cooled to room temperature to obtain a viscous solution (hereinafter abbreviated as "LMW12").

[0129] Example 1b. 100 g of the above solid lignin fraction (10% w / w) was hot mixed with 800 g of 1,3-propylene glycol and 100 g of NH4OH (30% solution). The mixture was cooled to room temperature and then filtered to give a black solution (hereinafter abbreviated as "LMW11").

[0130] Example 1c. 100 g of the above solid lignin fraction (10% w / w) was hot mixed with 835 g of 1,3-propylene glycol and 65 g of KOH (20% solution). The mixture was cooled to room temperature and then filtered to give a black solution (hereinafter abbreviated as "LMW10").

[0131] Example 2. The following lignin fractions were extracted from kraft black liquor, said lignin fractions having the following characteristics: >95% of total solids Single species: Southern pine M w : 4400-5000 Da (24-28 phenylpropane units) M n : 1200-1300 Da (6-7 phenylpropane units) OH group structure: Aliphatic 2.1mmol / g Carboxylic acid 0.5mmol / g Condensed and syringyl 1.7mmol / g Guaiacil 2.0mmol / g Catechol and p-OH-phenyl 4.0mmol / g

[0132] Example 2a. 100 g of the above lignin fraction (10% w / w) was hot mixed with 840 g of 1,3-propylene glycol and 60 g of NH4OH (30% solution). The mixture was cooled to room temperature and then filtered to give a black solution (hereinafter abbreviated as "OX11").

[0133] Example 2b. 100 g of the above lignin fraction (10% w / w) was hot mixed with 840 g of 1,3-propylene glycol and 60 g of NaOH (30% solution). The mixture was cooled to room temperature and then filtered to give a black solution (hereinafter abbreviated as "OX10").

[0134] Example 3. The antibacterial activity of the products of Example 1 was evaluated by in vitro antimicrobial susceptibility testing using the broth microdilution method (CLSI protocol - Clinical and Laboratory Standards Institute). The minimum inhibitory concentrations (MICs) of four products (blank, LMW12, LMW11, LMW10) were measured in multiwell plates. The blank was 1,3-propylene glycol only. The products were tested for antimicrobial activity against the following microorganisms (bacteria and fungi):

[0135] Specific Screening and Biocontrol bacteria - Escherichia coli - Pseudomonas aeruginosa - Staphylococcus aureus - Salmonella enteritidis - Campylobacter jejuni fungi - Candida albicans

[0136] result The compounds were tested neat and after adjusting the pH to 7 and 8. All experiments were performed in triplicate and yielded very similar inhibition results. The results are summarized in the table below.

[0137] [Table 1]

[0138] [Table 2]

[0139] Example 4. Determination of the minimum inhibitory concentration The minimum inhibitory concentrations (MICs) of the lignin fraction of Example 1b against five probiotic microorganisms were evaluated by antimicrobial susceptibility testing (in vitro testing) using the broth microdilution method (CLSI protocol - Clinical and Laboratory Standards Institute).

[0140] The microorganisms used were Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Enterococcus faecium, and Saccaromyces boulardii.

[0141] In all studies, positive controls for antibacterial activity were prepared using ceftriaxone (for bacteria) or fluconazole (for yeast). The results are summarized in the table below. Samples were diluted and evaluated for growth (+) and growth inhibition (-) at various dilutions. The grey squares indicate the minimum concentration (MIC) at which growth inhibition was observed.

[0142] [Table 3]

[0143] [Table 4]

[0144] [Table 5]

[0145] [Table 6]

[0146] [Table 7]

[0147] As described above, it is clear that the lignin fraction of the present invention does not have any adverse effects on probiotic microorganisms.

[0148] Example 5. Measurement of prebiotic activity The lignin fraction of Example 1b also exhibited prebiotic activity against E. faecium. The prebiotic activity of the lignin fraction of Example 1b was tested at concentrations below the relevant MIC obtained against the probiotic bacterium E. faecium.

[0149] [Table 8]

[0150] The prebiotic activity was evaluated by a quantitative formula (prebiotic index) comparing the growth of probiotic bacteria and enteric bacteria (E. coli, S. thyphimurium) in the presence of the lignin fraction of Example 1b. The prebiotic index was calculated as follows:

[0151]

number

[0152] In the formula, the first term represents the growth of probiotic bacteria and the second term represents the growth of enterobacteria.

[0153] Prebiotic compounds can selectively enhance the growth of certain probiotic bacteria as opposed to gut bacteria that do not use the compound to improve growth. Consequently, a prebiotic index >0 indicates prebiotic activity.

[0154] [Table 9]

[0155] Example 6. Use of lignin fractions in beef cattle feeding Experimental design, animal care, meat sampling and analysis The study involved 40 beef cattle, randomly divided into two groups: a control group and an experimental group. Both groups received the same feed ratio: commercial feed (calculated based on 2% of live weight) and ad libitum mixed hay. Prior to the start of the study, all animals were weighed and divided into two groups according to their live weights to achieve the same average live weight at the start of the study. The study lasted 120 days, 120 days before the animals were slaughtered. Unlike the control group, the experimental group received daily doses of the lignin fraction of Example 1b at 35g per animal for the first 90 days and 70g per animal for the next 30 days.

[0156] All animals were weighed monthly to assess final live weight, monthly weight gain, and food conversion index. Additionally, blood samples were collected throughout the study to assess blood counts and biochemical profiles.

[0157] Animals were transported and slaughtered at a European Community-approved abattoir in accordance with European Community law (1 / 2005EC) on animal welfare during transport and European Community regulation (1099 / 2009EC) on animal welfare for the slaughter of commercial animals. Immediately after slaughter, carcasses were stored at 4°C for 24 hours. Muscle samples (approximately 4 kg each) of the Longissimus thoracis et lumborum (LTL) muscle, located between the 13th and 18th thoracic vertebrae, were then taken for analysis.

[0158] The meat from each animal was stored at 4°C, and colorimetric parameters were measured during aging on days 3, 6, 9, and 14. Surface meat color was measured using a Minolta CR-300 colorimeter (illuminant D65; Minolta Camera Co., Ltd., Osaka, Japan) using the CIE L * , a * , b *(CIE, 1976) color system. Reflectance measurements were collected from a 0° viewing angle using an A-pulse xenon arc lamp with a reading surface of 8 mm diameter. For each meat sample, three measurements were taken at three different points. The three measurements at each point were obtained by rotating the detector system by 90° compared to the previous one, resulting in a total of nine measurements per sample. The colorimeter was equipped with a white title (L * =99.2, a * =1.0, b * =1.9) was used to calibrate the Hunter-Lab color space system. * and b * Using these values, saturation = (a² + b²)½ and hue (°) = tan-1(b / a) were calculated according to De Palo et al. ("Colour changes in meat of foals as affected by slaughtering age and post-thawing time", 2012, Asian-Australasian Journal of Animal Sciences, 25, 1775-1779).

[0159] Additionally, rheological parameters were measured. pH was recorded using a portable pH meter (Carlo Erba pH 710; Carlo Erba Reagenti, Milan, Italy) with a glass electrode designed to easily penetrate the meat. Before each measurement, the pH meter was automatically calibrated for muscle temperature using solutions with pH values ​​of 4 and 7 (Crison, Lainate, Italy). Water-holding capacity (WHC), cooking loss, and Warner-Bratzler shear force (WBSF) were measured as described by De Palo et al. ("Effect of nutrient level on carcass traits and meat quality of IHDH foals," 2014 Animal Science Journal, 85, 780-786).

[0160] Additionally, in vitro digestion was performed to assess total gas production over a 24-h period.

[0161] result Here, more interesting results are reported regarding meat quality, especially colorimetric profile, and in vitro digestive gas production.

[0162] In particular, as reported in Figure 1, redness (redness) was shown to differ between the two groups. Meat from beef cattle fed the lignin fraction exhibited higher redness throughout the entire aging period up to day 14. Recent studies have suggested that dietary effects on muscle color may be due to changes in glycogen stores, chilling rates, or antioxidant accumulation. All of these may ultimately be related to the basic intrinsic color characteristics of muscle. Normally, redness tends to decrease during aging, exhibiting a downward trend. This was not the case for meat samples from the experimental group. The decrease in redness on the surface of the meat is due to a biochemical phenomenon that occurs with exposure of myoglobin pigment to air. Oxidation of myoglobin causes meat darkening. In the experimental group, the antioxidant content of the animals' diets was hypothesized to result in higher redness of the meat and higher consumer ratings.

[0163] Figure 2 reports the trend in yellowness (yellowness). While control animals showed a downward trend in this indicator, antioxidant-treated animals showed no change in its value during aging, demonstrating a stable yellowness. Yellowness is closely related to the quantity and quality of intramuscular fat. Considering the same diet, similar weight gain and live weight at slaughter, and similar chemical composition of the meat, no differences in intramuscular fat mass were observed between the two groups. Instead, these differences may be due to the different quality of intramuscular fat with different fatty acid profiles. Indeed, differences in yellowness stability may be due to the progression of lipid oxidation processes, possibly following the release of lipolytic enzymes in the intercellular space, and redox activity within the fiber.

[0164] Figure 3 reports the 24-hour total gas production (ml) of in vitro ruminal fluid digestion. Animals in the experimental group produced lower gas at all times, producing 2864 ml compared to 3545 ml in the control group. This represents a surprising reduction in gas production of approximately 20%. Other studies have shown that some antioxidants used in dairy cow feeds reduce microbial replication and digestive activity. This, in turn, reduces digestive activity in the rumen and reduces both milk and meat production. Considering that live weight at slaughter, monthly weight gain, and dressing percentage were unchanged between groups, the reduction in gas production in the experimental group was unexpectedly not associated with a reduction in meat production. For this reason, the reduced gas production is likely methane, given that a reduction in volatile fatty acids should reduce meat and live weight conversion rates.

[0165] Example 7. Evaluation of livestock fertility and milk quality obtained from dairy cows fed the lignin fraction of the present invention The objective of this study was to use the lignin fraction of Example 1b to improve dairy herd health, improve economic performance and improve milk quality. Test duration: 28 days; 30 grams per cow per day

[0166] Test sample description: The Friesian herd consisted of an average of 85 lactating cows, 20 dry cows and associated company returns.

[0167] The cows were kept stable in comfortable structures made of straw bedding covered inside and outside by canopy structures.

[0168] Dry cows were housed in permanent facilities adjacent to straw bedding for the resting and calving area, a central paddock for walking and a covered shelter for the feeding zone.

[0169] All areas have ample space for resting and access to the manger. Companies that care about genetics that lead to good choices aim to improve the lifespan of their clothes and the quality of their milk.

[0170] Single flock unifeed: - Corn silage; - Lucerne hay (purchase); - Cornflower; - Permanent pasture hay; - Soybeans, cores; - Supplements and tampons.

[0171] 2014 production: 114 tonnes of milk per cow (AIA data source).

[0172] Herd health at the start of the study: The herd experienced a very harsh summer characterized by a very long warm period, approximately 50-60 days without rain, very high temperatures, and especially high humidity. Despite implementing all possible environmental conditioning systems, the cows reported various swallowing problems as a result: - Decrease in total milk production; - Reduction of casein and fat in milk; - Increased presence of lameness; - A gradual increase in cell count, reaching a peak of 577,000 cells; - Dairy cow fertility: negative impacts due to difficulty in manifesting estrus and low conception rates.

[0173] remarks: Trial: A lactating herd of 77 cows was integrated with the entire beef herd. The mean duration of lactation overall varied between 180 and 185 days. At the start of the study, the health of the herd was suboptimal: - Some cattle had just caught influenza, characterized by diarrhea and loss of appetite; - 7 cows have moderate severity of lameness.

[0174] The gradual introduction of the lignin fraction into the mixed unifeed was uneventful, and after the first week, intake gradually increased and stabilized.

[0175] Results: As certified by podiatrists, those animals who presented with influenza-related fatigue and lameness recovered quickly.

[0176] As intake increased, milk production increased immediately and the animals' body condition improved.

[0177] Somatic cell counts (SCC) were constantly monitored by the company, with one of the dairymen providing quality data. The initial SCC of 240,000 / 255,000 cells dropped to 120,000 in the second week and remained stable at approximately 140,000 / 150,000 cells until the end of the study.

[0178] Quick: Rebalancing of fat and casein in milk brought it back to more appropriate values ​​for the company. Improved general health of the cattle herd. Increased general fertility.

[0179] Milk production results: - Milk production started at 27.83 l / head on day 0 and reached a maximum of 33.41 l / head on day 35. - The increase in daily production per cow recorded on the 35th day after the start of administration of the lignin fraction was +20.08% compared to the initial production per cow. - Overall, the production increase was +138g / head / day. The largest absolute daily increase was recorded on days 16–17 (+2.43 l / head).

[0180] Milk quality performance: - Increased total polyphenol concentration in milk; - Increased milk production per cow per day; - Decreased somatic cell count in the acute phase; - Increased absorption capacity of free radicals (TEAC).

[0181] The results are highly significant, with a peak total polyphenol concentration of approximately 250%, an increase in overall milk production of +20.8%, a decrease in somatic cell count of -31%, and an increase in TEAC of +33%.

[0182] It should also be noted that these results showed a particular synchronization, with the maximum values ​​concentrated around the 15th to 17th day of treatment.

[0183] It was also found that supplementation according to the present invention produced a condition that extended the benefits for at least two weeks after cessation. The beneficial effects began to wane after about 20 days, as evidenced by a regrowth of somatic cells in the milk.

[0184] Since cow's milk usually contains only trace amounts of polyphenols, it is highly appreciated that milk obtained from cows fed the food supplement of the present invention unexpectedly contains up to 457 mg / l of polyphenols. Considering that the human requirement for polyphenols is 820 mg / die and that the average human intake of milk is 250 ml / die, the contribution of polyphenols provided by the milk of the present invention is 114 mg, when as stated, this is usually close to zero.

Claims

1. A supplement for improving the quality and shelf life of meat obtained from livestock, comprising: the supplement comprises a lignin fraction; the lignin fraction comprises fragments having a weight average molecular weight of up to 2,500 as measured by size exclusion chromatography; the fragments contain, on average by weight, up to 13 phenylpropane units; A supplement administered in an amount effective to improve the quality and shelf life of meat obtained from livestock.

2. Further, the lignin fraction contains fragments having a weight average molecular weight of 3,500 to 5,500 as measured by size exclusion chromatography; 2. A supplement for improving the quality and shelf life of meat obtained from livestock according to claim 1, wherein said fragments contain an average of up to 40 phenylpropane units by weight.

3. further comprising at least one resin acid; 3. A supplement for improving the quality and shelf life of meat obtained from livestock according to claim 1 or 2, wherein the resin acid is abietic acid, dehydroabietic acid, palustric acid, neoabietic acid, pimaric acid, isopimaric acid, sandalopimaric acid, or an ester thereof, or an ether thereof, or an alkali or alkaline earth salt thereof, or a mixture thereof.

4. Furthermore, C １２ ~C ２４ A supplement for improving the quality and shelf life of meat obtained from livestock according to any one of claims 1 to 3, comprising at least one alkali or alkaline earth salt of a fatty acid, at least one fatty oil, or a mixture thereof.

5. A supplement for improving the quality and shelf life of meat obtained from livestock according to any one of claims 1 to 4, used in an amount of up to 10 kg per tonne of animal feed, more preferably 1 to 5 kg per tonne of animal feed.

6. A supplement for improving the quality and shelf life of meat obtained from livestock described in any one of claims 1 to 4, administered to ruminants at a daily dose containing 20 to 100 g of lignin fraction.

7. A supplement for improving milk production and quality obtained from female livestock, comprising: the supplement comprises a lignin fraction; the lignin fraction comprises fragments having a weight average molecular weight of up to 2,500 as measured by size exclusion chromatography; the fragments contain, on average by weight, up to 13 phenylpropane units; A supplement administered in an amount effective to improve the quantity and quality of milk produced by female livestock.

8. Further, the present invention comprises a lignin fraction containing a fragment having a weight average molecular weight of 3,500 to 5,500 as measured by size exclusion chromatography, 8. A supplement for improving milk production and quality obtained from female livestock as described in claim 7, wherein said fragments contain a weight average of up to 40 phenylpropane units.

9. Further comprising at least one resin acid, 9. A supplement for improving milk production and quality obtained from female livestock according to claim 7 or 8, wherein the resin acid is abietic acid, dehydroabietic acid, palustric acid, neoabietic acid, pimaric acid, isopimaric acid, sandalopimaric acid, or an ester thereof, or an ether thereof, or an alkali or alkaline earth salt thereof, or a mixture thereof.

10. A supplement for improving milk production and quality obtained from female livestock described in any one of claims 7 to 9, further comprising at least one alkali or alkaline earth salt of a C12 to C24 fatty acid, at least one fatty oil, or a mixture thereof.

11. A supplement for improving milk production and quality obtained from female livestock described in any one of claims 7 to 10, used in an amount of up to 10 kg per tonne of animal feed, more preferably 1 to 5 kg per tonne of animal feed.

12. A supplement for improving milk production and quality obtained from female livestock described in any one of claims 7 to 10, administered to a ruminant at a daily dose containing 20 to 100 g of lignin fraction.

13. A supplement for improving the fertility of livestock, comprising: the supplement comprises a lignin fraction; the lignin fraction comprises fragments having a weight average molecular weight of up to 2,500 as measured by size exclusion chromatography; the fragments contain, on average by weight, up to 13 phenylpropane units; A supplement administered in effective amounts to improve the reproductive performance of livestock.

14. The present invention further comprises a lignin fraction containing fragments having a weight average molecular weight of 3,500 to 5,500 as measured by size exclusion chromatography, 14. The supplement for improving livestock fertility of claim 13, wherein the fragments contain a weight average of up to 40 phenylpropane units.

15. Further comprising at least one resin acid, 15. A supplement for improving livestock fertility according to claim 13 or 14, wherein the resin acid is abietic acid, dehydroabietic acid, palustric acid, neoabietic acid, pimaric acid, isopimaric acid, sandalopimaric acid, or an ester thereof, or an ether thereof, or an alkali or alkaline earth salt thereof, or a mixture thereof.

16. A supplement for improving livestock fertility according to any one of claims 13 to 15, further comprising at least one alkali or alkaline earth salt of a C12 to C24 fatty acid, at least one fatty oil, or a mixture thereof.

17. A supplement for improving livestock fertility according to any one of claims 13 to 16, used in an amount of up to 10 kg per tonne of animal feed, more preferably 1 to 5 kg per tonne of animal feed.

18. A supplement for improving livestock fertility as described in any one of claims 13 to 16, administered to ruminants at a daily dose containing 20 to 100 g of the lignin fraction.

19. A supplement for reducing methane emissions, comprising: the supplement comprises a lignin fraction; the lignin fraction comprises fragments having a weight average molecular weight of up to 2,500 as measured by size exclusion chromatography; the fragments contain, on average by weight, up to 13 phenylpropane units; A supplement administered in an amount effective to reduce methane emissions without impairing digestive efficiency or rumination.

20. The present invention further comprises a lignin fraction containing fragments having a weight average molecular weight of 3,500 to 5,500 as measured by size exclusion chromatography, 20. The supplement for reducing methane emissions of claim 19, wherein the fragments contain a weight average of up to 40 phenylpropane units.

21. Further comprising at least one resin acid, 21. A supplement for reducing methane emissions according to claim 19 or 20, wherein the resin acid is abietic acid, dehydroabietic acid, palustric acid, neoabietic acid, pimaric acid, isopimaric acid, sandalopimaric acid, or an ester thereof, or an ether thereof, or an alkali or alkaline earth salt thereof, or a mixture thereof.

22. The supplement for reducing methane emissions according to any one of claims 19 to 21, further comprising at least one alkali or alkaline earth salt of a C12 to C24 fatty acid, at least one fatty oil, or a mixture thereof.

23. A supplement for reducing methane emissions according to any one of claims 19 to 22, used in an amount of up to 10 kg per tonne of animal feed, more preferably 1 to 5 kg per tonne of animal feed.

24. A supplement for reducing methane emissions according to any one of claims 19 to 22, administered to a ruminant at a daily dose containing 20 to 100 g of the lignin fraction.

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