Animal feed additive compositions and methods of use thereof

Abstract

The present disclosure provides feed additives for domesticated animals capable of stimulating muscle function and health and increasing feed performance, as well as related methods for promoting muscle function and health, promoting growth and animal performance in domesticated animals. The animal feed additive provided by the invention contains Ribes alpinum or an extract thereof. The provided animal feed additives also contain lagopsis (lagopsis), related compounds, or biologically acceptable salts thereof, and in some embodiments, the total lignans are standardized with the content of furanguaiacol-like compounds and guaiacol-like compounds. The present disclosure also describes methods of improving muscle function and health by adding Ribes alpinum or an extract thereof to a diet, and methods of improving muscle function and health by adding Lagrenoline, related compounds, or a biologically acceptable salt thereof to a diet, and methods of improving muscle function and health by adding Lagrenoline, related compounds, or biologically acceptable salts thereof to a diet. Or a method for improving growth by standardizing total lignans in furanguaiacol compounds and guaiacol compounds, or in furanguaiacol compounds and guaiacol compounds.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application Serial No. 63 / 443,036, filed on February 2, 2023, the entire disclosure of which is incorporated herein by reference. Technical Field

[0003] The present disclosure relates to the field of animal feed additives, and more particularly, to compositions and methods for improving the performance of domestic animals. Background Art

[0004] The world's population is projected to double over the next 20 years, which will double the demand for food from animal sources. This challenge has been met before, as animal-source food production doubled in the second half of the 20th century. However, while the challenge is similar, the means to achieve the goal are very different. Indeed, the dramatic growth in livestock production during the 20th century was driven by advances in traditional nutrition, farm management, genetics, and the barebones use of antibiotics and chemical compounds. Today, bans on antibiotics in animal feed in some countries, restrictions on the use of therapeutic drugs at the farm level in others, and the global increase in non-traditional feeds in farm animal diets are undoubtedly altering the interaction between diet and farm animals, resulting in diets that are less nutritious than in the past. Even if demand for food from animal sources doubles, food production must respect animal welfare while also addressing consumers' growing expectations for environmentally safe and drug-free feed.

[0005] Thanks to a significant increase in genetic potential over the past 50 years, modern farm animals possess a remarkably robust metabolic capacity, fueled by their drive for rapid growth. Optimizing this metabolism by providing both nutrients and non-nutrients is a key role in the diets fed to these animals. Advances in husbandry, traditional nutrition, or scientific knowledge have already improved the production performance of these animals. For example, scientific advances in liver and, more recently, intestinal function have facilitated new approaches to farm animal dietary inputs. However, one organ remains underappreciated. Skeletal muscle is clearly the most valuable component of meat-producing animals. Furthermore, the role that muscle may play during animal development is not fully understood. For example, skeletal muscle consumes up to 25-30% of postprandial glucose, and skeletal muscle health is crucial for glucose and energy homeostasis. Therefore, for humans, improving skeletal muscle metabolic function and insulin sensitivity can have a significant impact on overall physiology and improve quality of life. Several factors prevalent in the farm animal environment are known to perturb skeletal muscle metabolic function, including oxidative stress and chronic inflammation. An imbalance between pro-inflammatory and anti-inflammatory cytokines can adversely affect muscle health. For example, elevated circulating pro-inflammatory cytokines such as TNFα induce insulin resistance and alter glucose homeostasis. This is exacerbated in metabolically robust farm animals with chronic inflammation (REF) or skeletal muscle, making them highly susceptible to oxidative stress.

[0006] Therefore, there is a great need in the art to develop feed additives that can replace antibiotics and other ingredients while improving the performance of farm animals in a drug-free environment. In addition, there is a great need in the art for feed additives that improve the muscle health of farm animals.

[0007] Summary of the Invention

[0008] In one embodiment, the present disclosure provides an animal feed ingredient composition comprising an effective amount of Ribes alpinum or an extract thereof, wherein the effective amount improves the performance of the animal feed to which the ingredient is added; and wherein the composition further comprises: a) an effective amount of furoguaiacin or a furoguaiacin-like compound or a guaiacin or a guaiacin-like compound, or a biologically acceptable salt thereof, wherein the effective amount improves the performance of the animal feed to which the ingredient is added; b) an effective amount of guaiacin or a related compound, including but not limited to guaiacin-like diterpenes such as marrubinic acid. acid, marrubenol, 12-S-hydroxymarrubiin, 11-oxo-marrubiin, premarrubiin, 3-deoxo-15-S-methocyvelutein, marrilibalacetal, cyllenin A, polydinonine, preleosibirin, or peregrinol, or a biologically acceptable salt thereof, wherein the effective amount improves the performance of the animal feed to which the ingredient is added; or c) an effective amount of a) and b). As used herein, "animal" refers to all animals other than humans. Examples of animals are monogastric animals and ruminants. Ruminants include, for example, sheep, goats, cattle, such as beef cattle, dairy cows, cows and calves, deer, camels, llamas, and kangaroos.Non-ruminant animals include monogastric animals such as pigs or swine (including but not limited to piglets, growing pigs and sows); poultry such as turkeys, ducks and chickens (including but not limited to broilers and laying hens); horses (including but not limited to hot-blooded, cold-blooded and warm-blooded horses), fish (including but not limited to amberjack, arapaima, barbel, sea bass, black carp, bocachico, bream, snakehead, giant pacu, carp, catfish, catfish, milkfish, salmon, cichlids, cod, croaker, dorada, totoaba, eel, goby, goldfish, threadfin, grouper, guapote, halibut, tuna, tilapia, tuna ... and crustaceans (including but not limited to shrimp and prawns). Animals that can be utilized according to the present invention include pets, including but not limited to dogs and cats.

[0009] Biologically acceptable salts useful in the present disclosure include, but are not limited to, acid addition salts formed with biologically acceptable acids, examples of which include hydrochloride, hydrobromide, sulfate or bisulfate, phosphate or biphosphate, acetate, benzoate, succinate, fumarate, maleate, lactate, citrate, tartrate, gluconate; methanesulfonate, benzenesulfonate and p-toluenesulfonate, alkali metal salts formed with bases, examples of which include sodium and potassium salts.

[0010] In certain embodiments, the animal feed provided herein comprises at least one animal feed component selected from the group consisting of vitamins, minerals, probiotics, enzymes, flavorings, amino acids, fats, essential oils, and preservatives. Non-limiting examples of vitamins include, for example, fat-soluble vitamins, including vitamin A, vitamin D3, vitamin E, and vitamin K, such as vitamin K3; and water-soluble vitamins, including vitamin B12, biotin, and choline, vitamin B1, vitamin B2, vitamin B6, niacin, folic acid, and pantothenic acid, such as Ca-D-pantothenic acid, and combinations thereof. Non-limiting examples of minerals include, for example, calcium, magnesium, potassium, and sodium, and trace minerals include boron, cobalt, chloride, chromium, copper, fluoride, iodine, iron, manganese, molybdenum, selenium, and zinc.Non-limiting examples of probiotics include, for example, Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus cereus, Bacillus pumilus, Bacillus polymyxa, Bacillus megaterium, Bacillus coagulans, Bacillus circulans, Bifidobacterium bifidum, Bifidobacterium animalis, Bifidobacterium sp., Carnobacterium sp., Clostridium butyricum, Clostridium sp., Enterococcus faecium, faecium), Enterococcus sp., Lactobacillus sp., Lactobacillus acidophilus, Lactobacillus farciminus, Lactobacillus rhamnosus, Lactobacillus reuteri, Lactobacillus salivarius, Lactococcus lactis, Lactococcus sp., Leuconostoc sp., Megasphaera elsdenii, Megasphaera sp., Pediococcus acidilactici, Pediococcus sp., Propionibacterium thoenii, Propionibacterium sp.) and Streptococcus sp. or any combination thereof.The limiting examples of enzymes include, for example, acetyl xylan esterase, acylglycerol lipase, amylase, α-amylase, β-amylase, arabinofuranosidase, cellobiohydrolase, cellulase, feruloyl esterase, galactanase, α-galactosidase, β-galactosidase, β-glucanase, β-glucosidase, lysophospholipase, lysozyme, α-mannosidase, β-mannosidase (mannanase), phytase, phospholipase A1, phospholipase A2, phospholipase D, protease, pullulanase, pectinesterase, triacylglycerol lipase, xylanase, β-xylosidase or any combination thereof. The limiting examples of flavorings include, for example, flower, berry, nut, caramel, chocolate, pepper, smoked, cheese or meat flavoring, mint such as peppermint, citrus flavoring such as orange and lemon, artificial vanilla, cinnamon and various fruit flavorings. Non-limiting examples of amino acids include, for example, alanine (Ala; A), arginine (Arg; R), asparagine (Asn; N), aspartic acid (aspartate, Asp; D), cysteine ​​(Cys; C), glutamine (Gln; Q), glutamic acid (glutamate; Glu; E), glycine (Gly; G), histidine (His; H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y) and valine (Val; V), and any combination thereof. Non-limiting examples of preservatives include, for example, sodium sorbate, potassium sorbate, sodium benzoate and potassium benzoate, and combinations thereof.

[0011] In some embodiments, the Ribes alpina composition is obtained from an extract of the plant or a part thereof. In further embodiments, the extract is produced from the stem, leaves, flowers, branches, roots, or fruit of the plant, or a part thereof, or any combination thereof. In still further embodiments, the extract is an extract of water, ethanol, methanol, isopropanol, ethyl acetate, acetone, or hexane, or a mixture thereof, or an extract of supercritical CO2.

[0012] In other embodiments of the compositions provided herein, the Ribes alpina or an extract thereof comprises about 5%-100% of the composition, including, for example, at least about 10% of the composition. In other embodiments, the Ribes alpina or an extract thereof in the composition comprises at least 10% of the composition.

[0013] In various embodiments, the amount of Ribes alpina or its extract is about 3 g to about 100 g, about 200 g, or about 500 g per metric ton of animal feed. In certain embodiments, the amount of Ribes alpina or its extract is about 100 mg per kg of animal feed.

[0014] In other embodiments, the present disclosure provides an animal feed comprising an animal feed ingredient composition as described herein, comprising an effective amount of Ribes alpina or an extract thereof, and further comprising a) an effective amount of furanoguaiacine or a furanoguaiac-like compound or guaiacine or a guaiac-like compound, or a biologically acceptable salt thereof, wherein the effective amount improves the performance of the animal feed to which the ingredient is added; b) an effective amount of summergrassin or a related compound as described herein, or a biologically acceptable salt thereof, wherein the effective amount improves the performance of the animal feed to which the ingredient is added; or c) an effective amount of a) and b) in an amount effective to improve the zootechnical performance of an animal fed the animal feed relative to a control feed lacking the animal feed ingredient composition. In certain embodiments, the animal feed is chicken feed, pig feed, dairy cow feed, or beef cattle feed. In specific embodiments, the improved zootechnical performance is improved muscle function and health or increased average daily weight gain, increased feed efficiency, or decreased feed conversion ratio.

[0015] As used herein, the term "feed conversion ratio" refers to the amount of feed fed to an animal that results in a certain amount of weight gain in the animal. An improved feed conversion ratio means a lower or decreased feed conversion ratio. A "lower or decreased feed conversion ratio" or "improved feed conversion ratio" means that the use of a feed additive composition in a feed results in a lower amount of feed required to feed the animal to increase the animal's weight by a specified amount, compared to the amount of feed required to increase the animal's weight by the same amount when the feed does not include the feed ingredient or additive composition.

[0016] The term "feed efficiency" refers to the weight gain per unit of feed when an animal is fed ad libitum or fed a prescribed amount of food over a period of time. "Improved feed efficiency" means that the use of the feed additive composition according to the present invention in a feed results in increased weight gain per unit of feed intake compared to a feed without the addition of the feed ingredient or additive composition.

[0017] In some embodiments, the composition is further defined as comprising furanoguaiac or a furanoguaiac-like compound or guaiac or a guaiac-like compound, or a biologically acceptable salt thereof. In certain embodiments, the furanoguaiac or a furanoguaiac-like compound or guaiac or a biologically acceptable salt thereof is present in an amount effective to improve the performance of the animal feed to which the ingredient is added relative to when the furanoguaiac or a furanoguaiac-like compound or guaiac or a biologically acceptable salt thereof is not present in the animal feed. In some embodiments, the furanoguaiac or furanoguaiac-like compound or guaiac or guaiac-like compound is obtained from Machilus edulis, Persea fructifera, Machilus thunberghii, Cinnamomum philippense, Saurus cernuus, Myristica fragrans, Guaiacum sanctum, or Guaiacum officinale plants or plant parts, or an extract thereof. In other embodiments, the furanoguaiac or furanoguaiac-like compound or guaiac or guaiac-like compound is obtained from the heartwood or branches of the Guaiacum sanctum or Guaiacum officinale plant, or an extract thereof. In other embodiments, the furanoguaiac or furanoguaiac-like compound or guaiac or guaiac-like compound is obtained from an extract of a plant or part thereof. In other embodiments, the extract is produced from the heartwood, whole trunk, bark, stem, leaf, flower, branch, root or fruit of a plant or part thereof, or any combination thereof. In further embodiments, the extract is an extract of water, ethanol, methanol, isopropanol, ethyl acetate, acetone, hexane or a mixture thereof, or an extract of supercritical CO2.

[0018] In certain embodiments, the furanoguaiac or furanoguaiac-like compound or guaiac or guaiac-like compound is synthetically prepared. In some embodiments, the furanoguaiac-like compound or guaiac-like compound has the following chemical structure:

[0019]

[0020] wherein R1 is -OH, -OCH3 or -OCH2, R2 is -OCH3 or -O, R3 is -OCH2 or -O, and R4 is -O or OCH3, and the corresponding glycoside forms. In other embodiments, the furanoguaiacine or furanoguaiacine-like compound or guaiacine or guaiacine-like compound corresponds to a peak with a retention time of about 12.5 minutes, 16.5 minutes, 18.8 minutes, 20.5 minutes, 26.3 minutes, 26.8 minutes, 34.7 minutes, 35.6 minutes, 36.8 minutes, 39.1 minutes, 41.9 minutes, 43.9 minutes, 44.6 minutes, 45.0 minutes, 46.3 minutes, 48.2 minutes, 50.7 minutes, 53.5 minutes, 54.6 minutes and 61.1 minutes on the HPLC chromatogram of the ethanol extract of the heartwood or branches of the guaiacine plant.

[0021] In certain embodiments, lignans comprise from about 0.5% to about 30% of the composition. In other embodiments, lignans comprise at least 9% of the composition. In further embodiments, the total lignans in the composition comprise at least 20% furanoguaiac or furanoguaiac-like compounds and guaiac or guaiac-like compounds. In further embodiments, the guaiac or guaiac-like compounds comprise from about 20% to about 60% by weight of the total lignans contained in the composition. In yet further embodiments, the furanoguaiac or furanoguaiac-like compounds comprise from about 20% to about 60% by weight of the total lignans contained in the composition.

[0022] In other embodiments, the aesculin or related compound is obtained from Marrubium vulgare, Marrubium velatinum, Marrubium cylleneum, Marrubium trachyticum, Marrubium globosum, Marrubium anisodon, Marrubium sericeum, Marrubium supinum, Leonotis leonurus, Leonotis nepetifolia, Phlomis bracteosa, Marrubium deserti de Noe, Marrubium alysson, or Marrubium thessalum plant or plant part, an extract thereof, or any combination thereof. In a specific embodiment, the aesculin or related compound is obtained from the aerial part of the Marrubium plant or an extract thereof. In certain embodiments, the aesculin or related compound is obtained from an extract of a plant, a part thereof, or any combination of its parts. In a further embodiment, the extract is produced from the stem, leaves, flowers, branches, roots or fruits of a plant, or parts thereof, or any combination thereof. In a further embodiment, the extract is an extract of water, ethanol, methanol, isopropanol, ethyl acetate, acetone or hexane, or mixtures thereof, or an extract of supercritical CO. In some embodiments, the scutellarin or related compound is synthetically produced. In other embodiments, the marrubiinic acid-related compound is marrubiinic acid, marrubenol, 12-S-hydroxymarrubiin, 11-oxo-marrubiin, premarrubiin, 3-deoxo-15-S-methocyvelutein, marrilibalacetal, cyllenin A, polydinonine, preleosibirin or peregrinol.

[0023] In certain embodiments, the aestivin has the following chemical structure:

[0024]

[0025] or a biologically acceptable salt thereof.

[0026] In other embodiments of the compositions provided herein, aestivin or a related compound comprises about 0.5%-100% of the composition, including, for example, at least about 0.7% of the composition. In some embodiments, the aesculus or related compounds comprise about 0.5% to about 99%, about 0.5% to about 95%, about 0.5% to about 90%, about 1% to about 80%, about 0.5% to about 75%, about 0.5% to about 70%, about 0.5% to about 60%, about 0.5% to about 50%, about 0.5% to about 40%, about 0.5% to about 30%, about 0.5% to about 25%, about 0.5% to about 20%, about 0.5% to about 15%, about 0.5% to about 10%, about 0.5% to about 7%, about 0.5% to about 5%, about 0.5% to about 2.5%, about 0.5% to about 1%, about 2.5% to 100%, about 5% to 100%, about 7%-100%, about 10%-100%, about 15%-100%, about 20%-100%, about 25%-100%, about 30%-100%, about 40%-100%, about 50%-100%, about 60%-100%, about 70%-100%, about 75%-100%, about 80%-100%, about 90%-100%, about 95%-100%, or about 99%-100%, about 0.7%-about 99%, about 1%-about 95%, about 2.5%-about 90%, about 5%-about 80%, about 7.5%-about 75%, about 10%-about 70%, about 20%-about 60%, about 25%-about 50%, or about 30%-about 40%. In other embodiments, the chelidonin or related compounds in the composition comprise at least 0.5%, 0.7%, 0.75%, 1%, 2.5%, 5%, 7%, 7.5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 95%, 99% or 100% of the composition.

[0027] In other embodiments, the present disclosure provides an animal feed ingredient composition comprising an effective amount of one or more plant parts from at least a first Herba Lysimachiae plant or an extract thereof, wherein the effective amount improves the performance of the animal feed to which the ingredient is added. In certain embodiments, the one or more plant parts comprise a whole plant. In some embodiments, the one or more plant parts comprise the stem, leaves, flowers, roots, fruits or cells of the plant, or any combination thereof. In other embodiments, the one or more plant parts comprise at least two different plant parts. In other embodiments, the one or more plant parts comprise plant parts from at least two different plants. In other embodiments, the one or more plant parts comprise plant parts from the same Herba Lysimachiae plant. In further embodiments, the one or more plant parts comprise plant parts from two or more different Herba Lysimachiae species. In yet further embodiments, the one or more plant parts are obtained from the above-ground parts of a Herba Lysimachiae plant or an extract thereof. In some embodiments, the one or more plant parts are ground into a powder or granules. In various embodiments, the marzipan plant is a species selected from the group consisting of Marrubium vulgare, Marrubium velatinum, Marrubium cylleneum, Marrubium trachyticum, Marrubium globosum, Marrubium anisodon, Marrubium sericeum, Marrubium supinum, Marrubium deserti de Noe, Marrubium alysson, or Marrubium thessalum plants, or wherein the composition comprises plant parts from any combination of species thereof.

[0028] In certain embodiments, the composition is further defined as comprising at least one animal feed component selected from the group consisting of vitamins, minerals, probiotics, enzymes, flavorings, amino acids, fats, essential oils, and preservatives, or any combination thereof. In some embodiments, the extract is an extract of water, ethanol, methanol, isopropanol, ethyl acetate, acetone, hexane, or a mixture thereof, or an extract of supercritical CO2. In other embodiments, the composition is further defined as comprising summergrass. In specific embodiments, summergrass comprises at least 0.5% of the composition. In other embodiments, summergrass comprises approximately 0.5%-100% of the composition.

[0029] In various embodiments, the amount of summergrass or related compounds or biologically acceptable salts thereof is about 0.5 mg to about 160 mg per kg of animal feed. In some embodiments, the amount of summergrass or related compounds or biologically acceptable salts thereof is about 0.5 mg to about 150 mg, about 0.5 mg to about 125 mg, about 0.5 mg to about 100 mg, about 0.5 mg to about 75 mg, about 0.5 mg to about 50 mg, about 0.5 mg to about 25 mg, about 0.5 mg to about 10 mg, about 0.5 mg to about 7.5 mg, about 0.5 mg to about 5 mg, about 0.5 mg to about 2.5 mg, about 0.5 mg to about 1 mg, about 0.5 mg to about 0.75 mg, about 1 In some embodiments, the amount of the herbicide-resistant phytoestrogens or related compounds or their biologically acceptable salts accounts for 1.6mg in every kg of animal feed. In other embodiments, the amount of summergrass or a related compound or a biologically acceptable salt thereof is about 0.5 mg, about 1 mg, about 2.5 mg, about 5 mg, about 7.5 mg, about 10 mg, about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg or about 160 mg per kg of animal feed.

[0030] In some embodiments, the aesculin or related compounds are obtained from Marrubium vulgare, Marrubium velatinum, Marrubium cylleneum, Marrubium trachyticum, Marrubium globosum, Marrubium anisodon, Marrubium sericeum, Marrubium supinum, Leonotis leonurus, Leonotis nepetifolia, Phlomis bracteosa, Marrubium deserti de Noe, Marrubium alysson, or Marrubium thessalum plants or plant parts, or extracts thereof. In other embodiments, the aesculin or related compounds are obtained from the above-ground parts of Marrubium plants, including but not limited to the stems, leaves, petioles, flowers, fruits, and seeds of Marrubium plants, or extracts thereof.

[0031] In yet another embodiment, the present disclosure provides a method for improving the performance of an animal feed, the method comprising adding to the animal feed an effective amount of a composition comprising Ribes alpina or an extract thereof as described herein, and further comprising an effective amount of furanoguaiacine or a furanoguaiac-like compound, or guaiacine or a guaiac-like compound or a biologically acceptable salt thereof, wherein the effective amount improves the performance of the animal feed to which the ingredient is added; an effective amount of marrubiin or a related compound, including but not limited to marrubiin-like diterpenes, such as marrubiinic acid, marrubenol, 12-S-hydroxymarrubiin, 11-oxo-marrubiin, premarrubiin, 3-deoxo-15-S-methocyvelutein, marrilibalacetal, cyllenin A, polydinonine, preleosibirin or peregrinol, or a biologically acceptable salt thereof, wherein the effective amount improves the performance of the animal feed to which the ingredient is added; or a component of the foregoing two. In certain embodiments, the improved performance comprises: improved muscle health and performance, altered feed intake, increased average daily weight gain, increased feed efficiency, decreased feed conversion ratio or increased milk production of animals fed the animal feed, relative to a control animal feed to which the animal feed ingredient composition is not added. In certain embodiments, the improved performance further comprises: altered feed intake, increased average daily weight gain, increased feed efficiency, decreased feed conversion ratio or increased milk production of animals fed the animal feed, relative to a control animal feed to which the animal feed ingredient composition is not added. In various embodiments, the animal feed is chicken feed, pig feed, dairy cow feed or beef cattle feed.

[0032] In yet another embodiment, the present disclosure also provides a method for improving the animal husbandry performance of an animal, the method comprising feeding the animal an animal feed comprising an animal feed ingredient composition, the animal feed ingredient composition comprising an effective amount of Ribes alpina or an extract thereof, the amount of which is effective to improve the animal husbandry performance of the animal fed the animal feed relative to a control feed lacking the animal feed ingredient composition. In other embodiments, the improved animal husbandry performance is altered feed intake, increased average daily weight gain, increased feed efficiency, decreased feed conversion ratio, or increased milk production. In a further embodiment, the animal feed comprises about 0.5 mg to about 160 mg of summergrass or a biologically acceptable salt thereof per kg of animal feed. In other embodiments, the amount of furanoguaiacine or furanoguaiac-like compound or guaiacine or guaiac-like compound or a biologically acceptable salt thereof is about 0.01 mg to about 2000 mg per kg of animal feed. In some embodiments, the animal is a chicken, a pig, a dairy cow, or a beef cattle. In other embodiments, the improved zootechnical performance is improved muscle health and performance, altered feed intake, increased average daily weight gain, improved feed efficiency, decreased feed conversion ratio, or increased milk production. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Those skilled in the art will understand that the drawings described below are for illustration purposes only and are not intended to limit the scope of the present invention in any way.

[0034] Figure 1 The ellagic acid concentrations of several Rubus and Ribes species are shown. The data show the mean ellagic acid concentrations in the species and their standard deviation.

[0035] Figure 2 Shown are muscle cell counts in C. elegans treated with a control and a Ribes alpina leaf extract. The Ribes alpina leaf extract, provided at 300 μg / ml, contained 0.0017 μg / ml ellagitannins, which, after hydrolysis, are expressed as ellagic acid. Bands with different letters are significantly different (P < 0.01).

[0036] Figure 3 Shown are the wiggling patterns of C. elegans 7 days after the L4 stage in a control group and a group treated with Ribes alpina leaf extract. The Ribes alpina extract provided at 300 μg / ml contained 0.0017 μg / ml ellagitannins, which are hydrolyzed to ellagic acid. Bands with different letters are significantly different (P < 0.02).

[0037] Figure 4This figure shows in vitro stimulation of protein production and myocyte proliferation in C2C12 cells by a 300 μg / mL Ribes alpina extract. The Y-axis shows protein content, used as an estimate of myocyte proliferation, expressed as a percentage of the control. The black bars represent the control without plant extract; the shaded bars represent the Ribes alpina extract-treated group. Standard deviations are shown above the bars.

[0038] Figure 5 Figure 3: Ribes alpina extract supplemented with increasing doses increases glucose uptake in C2C12 myocytes in vitro. The y-axis represents the response of the plant treatment (shaded bars) compared to the control (black bars). The standard deviation is shown above the bars.

[0039] Figure 6 The results show that 0.125 mg / mL and 0.25 mg / mL of Herba Ostreae extract reduced the release of the proinflammatory cytokine TNFα in human macrophage-like cells in vitro, measured as pg TNFα per μg protein. The control treatment group (black bars) was culture medium. The positive control group (white bars) was culture medium containing 1 μM dexamethasone. The values ​​shown in the figure are means. The bars show standard deviations.

[0040] Figure 7 The results show that 0.125 mg / mL and 0.25 mg / mL of Herba Ostreae extract reduced the release of the proinflammatory cytokine IL-6 in human macrophage-like cells in vitro, measured as pg of IL-6 per μg of protein. The control treatment (black bars) was culture medium. The positive control (white bars) was culture medium containing 1 μM dexamethasone. The values ​​shown in the figure are means. The bars indicate standard deviations.

[0041] Figure 8 The results show that 0.125 mg / mL and 0.25 mg / mL of Herba Ostreae extract reduced the release of the proinflammatory cytokine IL-1β in human macrophage-like cells in vitro, measured as pg of IL-1β per μg of protein. The control treatment (black bars) was culture medium. The positive control (white bars) was culture medium containing 1 μM dexamethasone. The values ​​shown in the figure are means. The bars represent standard deviations.

[0042] Figure 9 This figure shows that addition of 25 μg / mL of Herba Lycopersici extract stimulated protein production and myocyte proliferation in C2C12 cells in vitro. The Y-axis (expressed in μg / mL) shows protein content, used to assess myocyte proliferation. Dark bars represent controls without the addition of plant extract; shaded bars represent Herba Lycopersici treatments. Standard deviations are shown above the bars.

[0043] Figure 10This figure shows that the addition of 300 μg / mL of Herba Lysimachiae extract increases glucose uptake in C2C12 myocytes in vitro. The y-axis shows the response of the plant treatment group (shaded bars) compared to the control (set at 100%; black bars). The standard deviation is shown above the bars.

[0044] Figure 11 This figure shows that the extract of Osmanthus fragrans partially reverses the toxicity of 1 μM aflatoxin B1 on HepG2 cells in an in vitro bioassay. The black bar represents the untreated control group; the white bar represents the treatment group containing 1 μM aflatoxin B1; the shaded bar shows the treatment group with 1 μM aflatoxin B1 and increasing amounts of Osmanthus fragrans extract.

[0045] Figure 12 This figure shows that 300 μg / mL of Herba Lycoris radiata extract reduces PGE2 production in chondrocytes in an in vitro cell-based bioassay. The black, white, and shaded bars represent the control, positive control, and Herba Lycoris radiata extract-treated groups, respectively. Standard deviations are shown above the bars.

[0046] Figure 13 This figure shows that 300 μg / mL of Herba Lycoris radiata extract reduces matrix metalloproteinase-3 in chondrocytes in an in vitro cell-based bioassay. Black, white, and shaded bars represent the control, positive control, and Herba Lycoris radiata extract-treated groups, respectively. Standard deviations are shown above the bars.

[0047] Figure 14 This study demonstrates that supplementing a diet with 157 ppm of the aboveground portion of summer solstice grass improved feed efficiency, as measured by feed conversion ratio, in chickens, compared to both negative and positive controls. The negative control was a diet without the feed additive. The positive control was the same diet supplemented with a technology considered a standard alternative to antibiotic growth promoters. The values ​​shown are averages.

[0048] Figure 15The HPLC chromatogram of GS heartwood is shown. The numbers on the peaks are related to the compounds and their specific meanings are explained. The retention times (min) of the peaks are shown in brackets. Peaks 1 (5.519), 3 (15.576), 6 (19.474) and 8 (24.551): vanillins; Peaks 2 (12.480) and 17 (43.972): guaiaretic acids; Peaks 4 (16.694), 9 (26.295), 10 (26.810), 11 (35.346), 12 (35.888), 13 (37.983), 14 (39.385), 16 (41.869), 19 (45.074), 21 (48.2 Peaks 15 (41.336), 22 (49.639), 26 (57.693), 27 (61.201), 28 (64.556) and 30 (68.034): unknown compounds.

[0049] Figure 16 HPLC chromatogram of a branch of a mature tree (trunk diameter: 40 cm, branch diameter: 2.54 cm) is shown. The numbers on the peaks are related to the compounds and their specific meanings are explained. The retention time (min) of the peaks is shown in brackets. Peak 1 (5.513), 3 (15.597), 6 (19.493) and 8 (25.332): vanillins; Peak 2 (12.476) and 17 (43.981): guaiaretic acids; Peak 4 (16.112), 9 (26.340), 10 (26.872), 11 (33.806), 12 (35.386), 13 (36.421), 14 (37.887), 16 (41.887), 19 (43.861), 21 (48.2 Peaks 5 (18.840), 7 (20.581), 18 (44.586), 20 (45.090), 23 (50.674), 24 (53.558) and 29 (67.079): guaiacins; Peaks 15 (41.372), 22 (49.445), 26 (57.715), 27 (61.268), 28 (64.576) and 30 (68.015): unknown compounds.

[0050] Figure 17 Presentation of the total lignan content in % of different plant parts of Guaiac wood.

[0051] Figure 18The contents of furanoguaiacins, guaiacs, and guaiaretic acids in the heartwood, branches, sapwood, and bark of the sacred guaiac wood are shown. The contents are expressed as a percentage of the total lignans.

[0052] Figure 19 The results showed that the supplementation of the diet with 8 ppm of standardized furanoguaiacs and total lignans of guaiacs (diet 3-50% furanoguaiacs and 40% guaiacs) improved the feed conversion rate of chickens compared with the negative and positive controls. The negative control was a diet without feed additives (diet 1). The positive control was the same diet with the addition of a standard technology that is considered to be an alternative to antibiotic promoters (diet 2-100 ppm of Evolution-B, Code X60-6930). The values ​​shown in the figures are averages. In the figures, different letters on the bars indicate significant differences.

[0053] Figure 20 Shown are the effects of standard doses of total lignans, normalized to furanoguaiacins and guaiacinoids, on the production of the inflammatory cytokine TNFα. Negative controls consist of culture medium. Positive controls contain 1 μM dexamethasone. Values ​​shown are mean values. Bars also indicate standard deviations.

[0054] Figure 21 Shown are the effects of standard doses of total lignans, normalized to furanoguaiacins and guaiacinoids, on the production of the inflammatory cytokine IL-6. Negative controls consist of culture medium. Positive controls contain 1 μM dexamethasone. Values ​​shown are mean values. Bars also indicate standard deviations.

[0055] Figure 22 Shown are the effects of standard doses of total lignans, normalized to furanoguaiacins and guaiacinoids, on the production of the inflammatory cytokine IL-1β. Negative controls are indicated by culture medium. Positive controls included 1 μM dexamethasone. Values ​​shown are mean values. Bars also indicate standard deviations.

[0056] Figure 23 This figure illustrates the effects of standard doses of total lignans, standardized with furanoguaiacins and guaiacinoids, on PGE2 production in chondrocytes in an in vitro cell-based bioassay. Black and shaded bars represent the control and experimental treatment groups, respectively. Standard deviations are shown above the bars.

[0057] Figure 24 Figure 2 illustrates the effects of standard doses of total lignans, standardized with furanoguaiacins and guaiacinoids, on matrix metalloproteinase-3 in chondrocytes in an in vitro cell-based bioassay. Black and shaded bars represent the control and experimental treatment groups, respectively. Standard deviations are shown above the bars.

[0058] Figure 25 Displays the average bird weight results from 0-14 days.

[0059] Figure 26 FCR results adjusted for mortality on days 0-14 are shown.

[0060] Figure 27 Average body weight results for days 0-28 are shown.

[0061] Figure 28 FCR results adjusted for mortality from day 0 to 28 are shown.

[0062] Figure 29 Mean body weight results for days 0-42 are shown.

[0063] Figure 30 FCR results adjusted for mortality from days 0 to 42 are shown. DETAILED DESCRIPTION

[0064] The present disclosure provides feed additives for domestic animals that can stimulate muscle function and health, and related methods for promoting muscle function and health and zootechnical performance of domestic animals. The feed additives for domestic animals described herein can also increase feed performance, and related methods for promoting growth and zootechnical performance of domestic animals. The animal feed additives provided herein contain alpine tea leaf or an extract thereof, and in certain embodiments contain summer grass linalool, the content of which is standardized to total lignans of furanoguaiac compounds and guaiac compounds; in further embodiments, summer grass linalool and total lignans standardized to the content of furanoguaiac compounds and guaiac compounds. The present disclosure also describes a method for improving muscle function and health by adding alpine tea leaf or an extract thereof to the diet; and a method for improving growth by adding summer grass linalool to the diet, and in some embodiments, by adding total lignans standardized to furanoguaiac compounds and guaiac compounds to the diet.

[0065] Ellagitannins are hydrolyzable polyphenols with beneficial effects on human health. In the gastrointestinal tract, ellagitannins are hydrolyzed to ellagic acid, which is subsequently metabolized by the colonic microbiota into urolithins. These are referred to herein as ellagic acid / urolithins. Ellagitannins / urolithins have various health-promoting effects, such as inhibiting proliferation, combating inflammation, optimizing lipid metabolism, and inducing autophagy in cancer cell models. Animal studies have also demonstrated positive effects on mitochondrial and muscle function.

[0066] The present disclosure provides exemplary embodiments of novel compositions for inclusion in the diets of farm animals to increase protein synthesis, muscle mass, and / or muscle strength. The examples describe the addition of a plant, Ribes alpina, or an extract thereof, to the animal's diet to improve performance, muscle growth, and muscle health.

[0067] Ellagic acid / urolithin are known to have an effect on muscle growth. For example, urolithin assays in C. elegans showed that a concentration of 50 μM (11.4 μg / ml) was required to obtain a positive effect on muscle function. Furthermore, a concentration of 11.4 μg / mL of urolithin was required to trigger a muscle function response. Combinations of urolithin with protein sources, medium-chain triglycerides, or nicotinamide riboside have been suggested for the treatment of muscle-related pathological conditions. Compositions containing urolithin have also been suggested for increasing muscle cell size. In summary, ellagic acid / urolithin are clearly capable of promoting muscle function and muscle growth.

[0068] Urolithin production can be initiated by adding ellagitannins to the diet consumed by the subject, which will be digested into ellagic acid and subsequently fermented into urolithins. Common dietary sources of ellagitannins are fruits, nuts, leaves, and seeds of certain plant species. For example, urolithin levels are higher in pomegranates and berries belonging to the genera Rubus and Ribes. Based on the results described above, at least 11.4 mg / mL of urolithin is required to promote muscle cell health and function.

[0069] However, the present disclosure surprisingly encompasses plants that contain no or very low levels of ellagic acid / urolithins.

[0070] Farm animals are exposed to a variety of stressors, especially in today's production environment where antibiotics, drugs and chemicals are increasingly being banned from being used to feed these animals. For example, chickens are susceptible to chronic enteric inflammation (REF). It is now widely accepted that the use of antibiotics and other drugs is a "crutch" to reduce animals' response to various stressors. A new approach to improving the performance of farm animals is by reducing the impact of the various stressors they are exposed to. This holistic approach to improving performance not only considers, for example, direct improvements in daily weight gain or feed conversion or milk production, but also considers improvements in multiple responses that together improve the animal's condition, welfare and ultimately its output. It is against this background that the present invention proposes a new method for increasing animal productivity in a natural way while maintaining animal welfare. The addition of summer grass extract or the plant summer grass to the diet can improve several physiological problems faced by farm animals. The invention will be described in detail in the following examples.

[0071] Summer solstice herbicide compounds

[0072] The chemical structure of summer grass (C 20 H28 O4; structure 1 ) as shown below.

[0073]

[0074] The active compounds related to oxalin include but are not limited to oxalic acid (structure 2 ) and phytosporin (structure 3 ).

[0075]

[0076]

[0077] Marigold is a well-known diterpene lactone that constitutes the bitter ingredient of horehound and many other medicinal plants of the Lamiaceae family. It is one of the main components of Marigold, lion's ear flower and schizonepeta lion's ear flower. It is the main component of many Marigold species (Lamiaceae), including about 97 species distributed in temperate regions along the Mediterranean and Eurasia. Marigold is also present in plants including but not limited to velvet Marigold, Marrubium cylleneum, Marrubium trachyticum, spherical Marigold, Marrubium anisodon, Marrubium sericeum, false sedge, Siegesbeckia scabra, Marrubium deserti de Noe, garden mustard Marigold or Marrubium thessalum.

[0078] Phenolic compounds are a class of plant secondary metabolites that play an important role in plant physiology. Phenolic compounds vary widely in structure and can be divided into four groups based on their structure: phenolic acids, flavonoids, stilbenes, and lignans. Lignans are widely distributed throughout plants, such as cereals, grains, berries, and garlic. Total lignans refers to the sum of lignans in a plant composition.

[0079] The present disclosure provides a novel method for using total lignans, standardized with two specific lignan components (furoguaiacoid compounds and guaiacoid compounds), for improving the production performance (increased intake, daily weight gain and / or improved production efficiency) of farm animals or other domesticated animals. These compounds can be used under normal feeding conditions.

[0080] The term lignans covers a variety of chemical structures. Pilkington divides lignans into two main categories: classical lignans and neolignans, and two smaller categories: flavonolignans and coumarin lignans (Lignans: A Chemometric Analysis. Molecules 23: 1666, 2018). Each of these categories has several subcategories. Classical lignans have 6 main subcategories: CL1: dibenzylbutane, CL2: dibenzylbutyrolactone, CL3: arylnaphthalene / aryltetralin, CL4: dibenzocyclooctadiene, CL5: substituted tetrahydrofuran, and CL6: 2,6-diarylfurofuran. There are 15 subclasses of neolignans, the most common of which are: NL1: benzofuran, NL2: 1,4-benzodioxane, NL3: alkyl aryl ether, NL4: biphenyl, NL5: cyclobutane, NL6: 8-10-bicyclo[3.2.1]octane, NL7: 8-30-bicyclo[3.2.1]octane and NL8: biphenyl ethers.

[0081] The chemical structure of natural molecules is related to their function. Molecules with different chemical structures exhibit different biological or physiological functions across classes and even subclasses. Lignans from different lignan classes and subclasses exhibit different physiological functions and effects, and therefore, different performance effects on target animals. In other words, the physiological functions of lignans, and thus the performance responses of animals, are related to their chemical structure.

[0082] Furanoguaiac and guaiacine

[0083] Furanoguaiac (also known as α-guaiaretic acid, structure 4 ) is a classic lignan belonging to the subclass of substituted tetrahydrofurans (see subclass CL5 above) with the molecular formula C 20 H 20 O5.

[0084]

[0085] Guaiacin (Structure 5 ) is a classic lignan belonging to the arylnaphthalene / aryltetralin subclass of lignans (see the above-mentioned CL3 subclass), and its molecular formula is C 20 H 24 O4.

[0086]

[0087] Holy guaiac wood or guaiac wood is a source of guaiac or guaiac-like compounds. Guaiac gum or guaiac resin can be extracted from the heartwood of guaiac wood and holy guaiac wood. Furanoguaiac or furanoguaiac-like compounds and guaiac or guaiac-like compounds are primarily found in the guaiac wood or its extracts, although other sources of furanoguaiac or furanoguaiac-like compounds and guaiac or guaiac-like compounds include, but are not limited to, Machilus edulis, Persea fructifera, Machilus thunberghii, Cinnamomum philippense, Saurus cernuus, and Myristica fragrans.

[0088] Furanoguaiacins and guaiacine compounds

[0089] In certain embodiments, furanoguaiacins and guaiacin compounds may be represented by the core structure (Structure 6 ) to define:

[0090]

[0091] In certain embodiments, the R1, R2, R3, and R4 groups are as shown in Table 1 below.

[0092] Table 1

[0093] <![CDATA[R1]]> <![CDATA[R2]]> <![CDATA[R3]]> <![CDATA[R4]]> OH <![CDATA[OCH3]]> <![CDATA[OCH2]]> O <![CDATA[OCH3]]> <![CDATA[OCH3]]> <![CDATA[OCH2]]> O <![CDATA[OCH3]]> <![CDATA[OCH3]]> OH <![CDATA[OCH3]]> <![CDATA[OCH2]]> O <![CDATA[OCH2]]> O

[0094] In other embodiments, the furanoguaiacins and guaiacins include the corresponding glycoside forms.

[0095] Animal feed ingredients

[0096] Provided herein is a novel animal feed composition having improved performance due to the inclusion of an animal feed ingredient composition as described herein. Different domestic animals have different feed requirements. For example, the main ingredient in chicken feed is typically cereals, including but not limited to wheat, corn, sorghum, oats, barley or rye, protein (which can be from oilseed meal), and fat or oil. However, other ingredients can be added to chicken feed, including but not limited to calcium sources, salts, minerals, probiotics, vitamins, amino acids, flavorings and preservatives. Exemplary chicken feeds for chickens and pullets include protein, lysine, methionine, fat, fiber, calcium, phosphorus, sodium chloride, manganese, vitamin A and vitamin E.

[0097] The main ingredients in pig feed are usually grains, including but not limited to rice bran, broken rice and corn, protein, which can come from oilseed meal such as alfalfa meal or soybean meal, minerals and vitamins. However, other ingredients can be added to pig feed, including but not limited to calcium sources, salts, minerals, probiotics, vitamins, amino acids, flavorings and preservatives. Exemplary pig feeds include energy (grains), protein, vitamins, minerals, fiber, probiotics and plant ingredients.

[0098] The main ingredients in dairy cow feed are usually grains, protein, which can come from oilseed meal such as cottonseed meal or soybean meal, sugar and fat. However, other ingredients can be added to dairy cow feed, including but not limited to fiber, calcium sources, salt, minerals, probiotics, vitamins, amino acids, flavorings and preservatives.

[0099] The primary ingredients in beef cattle feed are typically grains, including but not limited to wheat, corn, sorghum, oats, barley, or rice, and protein, which can come from de-oiled rice bran, polished rice, wheat bran, or corn bran. However, other ingredients may be added to beef cattle feed, including but not limited to fiber, fat, salt, minerals, probiotics, vitamins, amino acids, flavorings, and preservatives.

[0100] The main ingredients in sheep feed are usually grains, such as alfalfa and corn, vitamins, such as vitamin A, vitamin D and vitamin E, selenium, inorganic salts and phosphorus. However, other ingredients can be added to sheep feed, including but not limited to fats, probiotics, amino acids, flavorings and preservatives. An exemplary sheep feed for adult ewes and rams includes grains, protein, fat, fiber, calcium, ammonium chloride, phosphorus, sodium chloride, selenium and vitamin A.

[0101] The main ingredients in goat feed are usually grains such as hay, alfalfa, barley, corn and oats; protein, which can come from distiller's grains and meal; fat; fiber and minerals such as calcium, phosphorus, NaCl, copper, selenium and vitamins such as vitamin A, vitamin D and vitamin E. However, other ingredients can be added to goat feed, including but not limited to probiotics, amino acids, flavorings and preservatives. Exemplary goat feeds include grain products, protein, fat, fiber, acid detergent fiber and calcium, phosphorus, NaCl as well as copper, selenium and vitamins such as vitamin A, vitamin D and vitamin E.

[0102] As described herein, the performance of any such animal feed can be improved by adding the animal feed ingredients provided by the present disclosure. The amount added can be optimized based on the type of feed, animal physiology, the conditions under which the animal is raised, and other conditions understood by those skilled in the art based on the teachings of the present disclosure.

[0103] Example

[0104] Example 1

[0105] The leaves of various berry varieties were analyzed for ellagic acid content by HPLC. Leaf samples were extracted to obtain ellagitannins, which were then hydrolyzed to produce ellagic acid. The ellagic acid content was determined using HPLC with ellagic acid as an external standard.

[0106] Figure 1 An overview of the ellagic acid content after hydrolysis is given. The ellagic acid content in leaves of different blackberry (Rubus fruticosus) varieties ranged from 0.68 to 1.65 mg / 100 g dry weight. In gooseberry leaves, the content ranged from 6.9 to 19.4 mg / 100 g. Higher levels, ranging from 6.6 to 32.5 mg / 100 g, were observed in samples of blackcurrant (Ribes nigrum). The highest absolute ellagic acid content, 146.6 mg / 100 g, was detected in samples of goldencurrant (Ribes aureum) and fragrantcurrant (R. odoratum (syn.)), respectively. The lowest ellagic acid content was found in alpine currant (0.188 mg / 100 g), with four of the five samples containing only trace amounts of ellagic acid. This analysis confirms that alpine currant, despite being a member of the genus Ribes, contains only trace amounts of ellagic acid.

[0107] Example 2

[0108] The potential effects of alpine currant on muscle growth were tested in the Caenorhabditis elegans model. Alpine currant extract was provided as a solution in 50% ethanol at a concentration of 300 μg / mL. This extract contained 0.58 μg / 100 mg of ellagitannins (expressed as ellagic acid after hydrolysis). Assuming complete conversion of ellagic acid to urolithins, the urolithin concentration in C. elegans treated with alpine currant extract was only 0.0017 μg / mL.

[0109] In all experiments, worm growth medium (NGM) supplemented with extracts was prepared as follows: a single aliquot of the sample was thawed in a 45°C water bath for 15 minutes. The completely dissolved sample was filtered through a 0.2μm cellulose acetate membrane sterile filter into a clean 1.5mL Eppendorf tube. Each extract was added to NGM to a final concentration of 300μg / mL and poured into a culture dish. Relevant controls included a negative control and an ethanol solvent control, 0.15% and 0.075%, respectively. Under all conditions, NGM was also prepared containing 100μM 5-fluoro-2'-deoxyuridine (FuDR) to prevent offspring from hatching. Age-synchronized worms were placed in NGM supplemented with compounds from the L1 to L4 stage and their motility and muscle cell number were assessed. When in the L4 stage, the worms were transferred to the compound culture dish. The animals' motility and muscle morphology were further assessed 7 days after L4 (after the end of the reproductive period).

[0110] Assessment of myocyte proliferation. Use worm strain AW306, wild-type worms expressing nuclear markers for body wall myocyte nuclei. Worms were cultured from L1 to L4 on each compound plate. When worms reached the L4 stage, the number of myocyte nuclei in the right abdominal quadrant (n=40) was counted.

[0111] Worm motility was assessed. Wild-type worms (N2 strain) were placed in M9 buffer for each condition (20-25 worms) and video was captured using a Leica S8aP0 binocular microscope with a Leica DMC2900 camera and LAS v4.12 software. Movies were analyzed using ImageJ v1.53. The number of body bends was quantified using the wrMTrck plugin for ImageJ (build 110622). The number of body bends per minute (BBPM) was calculated, and graphs were generated using GraphPad Prism showing the mean and standard deviation of the mean, with each point representing a single worm count.

[0112] The following results were observed. Ribes alpina extract containing 0.0017 μg / ml ellagitannins (expressed as ellagic acid after hydrolysis) showed a positive effect on muscle growth by promoting muscle proliferation in the L4 stage of Caenorhabditis elegans ( Figure 2 Ribes alpina extract, containing 0.0017 μg / ml ellagitannins (expressed as ellagic acid after hydrolysis), positively affected muscle function in C. elegans by increasing muscle oscillation 7 days after L4 ( Figure 3Considering that the lowest urolithin concentration that produces a muscle function response is 11.4 mg / mL urolithin, as detailed above, the amount of ellagic acid / urolithin added is 1.49123E-07 less than this lowest dose. This extremely low dose of ellagic acid / urolithin failed to promote muscle growth. Surprisingly, 0.3 mg of Ribes alpina leaf extract, containing 0.0017 μg / mL ellagic acid / urolithin, showed a positive effect on muscle growth. Therefore, it can be concluded that the effect of Ribes alpina on muscle growth is not caused by ellagitannins.

[0113] Example 3

[0114] This example evaluates the effects of Ribes alpina extract on muscle cell physiology in a cell-based bioassay.

[0115] Bioassays were performed using the mouse myoblast cell line C2C12. Ribes alpina extract was tested in triplicate at different concentrations. Two outcomes were measured. First, the myocyte proliferation assay aimed to investigate the effects of 300 μg / ml Ribes alpina extract on myocyte proliferation and growth. Determination of protein content was used to characterize cell proliferation in the applied in vitro model. 24 hours after inoculation, the C2C12 mouse myoblast cell line was supplemented with Ribes alpina extract for the first time. Five days later, cell differentiation was initiated. 24 hours after the start of differentiation, the second supplementation with Ribes alpina extract was performed. On day 3 of the differentiation phase, the final supplementation with Ribes alpina extract was performed. Seven days after the start of differentiation, the cells were lysed and the protein content was determined using the Bradford assay. A culture medium control was included for each batch. The assay was repeated three times. Second, the glucose uptake assay aimed to investigate the effects of Ribes alpina extract on glucose uptake in myocytes. C2C12 cells were inoculated and grown to confluence (3 days). Cells were differentiated for 4 days to form myotubes and then treated with Ribes alpina extract for 24 hours. Following supplementation, glucose was depleted for 24 hours, followed by serum depletion overnight. After washing, the fluorescent glucose analog 2-NBDG (2-(N-(7-nitrobenzo-2-oxa-1,3-diazol-4-yl)-amino)-2-deoxyglucose) was added to the culture medium to monitor glucose uptake into living cells. After 30 minutes, fluorescence was measured after washing and cell lysis. n = 6 cells were assayed.

[0116] The results showed that 300 μg / mL of Ribes alpina extract increased myocyte proliferation ( Figure 4 ), 30 μg / ml increased glucose uptake ( Figure 5). These results confirm that the muscle growth response of Ribes alpina is not mediated by ellagic acid / urolithins. In this example, a cell-based assay using C2C12 cells was used. It is known that only intestinal bacteria can convert ellagic acid into urolithins (REF). It is well known that this cell-based bioassay cannot observe the effects of components that require microbial biotransformation to be observed. Therefore, the effect of Ribes alpina extract on muscle cell proliferation and function is independent of ellagic acid, which needs to be converted into urolithins by intestinal microorganisms.

[0117] In summary, adding a small amount of alpine currant to the diet of farm animals can help promote muscle health and function. In some embodiments, up to 500g of alpine currant plant can be added per metric ton of feed, while in other embodiments, the dosage is 100-200g of plant per metric ton of feed.

[0118] Example 4

[0119] This example evaluates the effect of increasing doses of Herba Ostreae extract on anti-inflammatory cytokines in vitro. The Herba Ostreae plant extract (referred to as Herba Ostreae extract) was prepared as follows. Prior to extraction, the Herba Ostreae plant was ground in a centrifugal grinder. Approximately 100 g of ground Herba Ostreae was extracted with 1000 g of solvent at 40° C. for 60 min under stirring. The crude extract was then filtered under vacuum, and the filtrate was concentrated under reduced pressure in a rotary evaporator at 40° C. to a concentrated fluid extract having a dry matter content of 10%-35%.

[0120] The anti-inflammatory effect of 70% v / v ethanol Herba Sophorae flavescentis extract standardized with Herba Sophorae flavescentis was determined using the THP1 human monocyte cell line, which was differentiated into macrophage-like cells by phorbol myristate acetate. Prior to the assay, mitochondrial dehydrogenase activity in living cells was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay to determine the potential cytotoxic effects of the test compounds. The anti-inflammatory effect on cytokine release in THP-1 macrophages was measured by incubating the cells with three concentrations of Herba Sophorae flavescentis extract and then adding LPS to stimulate cytokine production. After 24 hours, the amount of TNF-α, IL-1β, and IL-6 in the cell culture supernatant was determined by capture ELISA using a commercially available kit.

[0121] The main results are summarized. Two doses of Herba Oleraceae extract (0.125mg / mL and 0.25mg / mL) reduced the production of inflammatory cytokine TNFα ( Figure 6 ), the production of inflammatory cytokine IL6 ( Figure 7) and the production of inflammatory cytokine IL1β ( Figure 8 ).

[0122] Example 5

[0123] This example evaluated the effect of a dose of a Herba Lycopodii extract on muscle cell physiology.

[0124] Bioassays were performed using the mouse myoblast cell line C2C12. The Herba Lysimachiae extract was tested in triplicate at concentrations of 25 and 30 μg / ml. Two results were measured. First, the effect of Herba Lysimachiae extract (25 μg / ml) on myocyte proliferation and growth was investigated using a myocyte proliferation assay. Determination of protein content was used to identify cell proliferation in the applied in vitro model. 24 hours after inoculation of the mouse myoblast cell line C2C12, Herba Lysimachiae extract was supplemented for the first time. Five days later, the cells began to differentiate. 24 hours after the start of differentiation, Herba Lysimachiae extract was supplemented for the second time. On day 3 of the differentiation phase, Herba Lysimachiae extract was supplemented for the final time. Seven days after the start of differentiation, the cells were lysed and the protein content was determined using the Bradford assay. A culture medium control was set up for each batch. The assay was repeated three times. Secondly, a glucose uptake assay was used to investigate the effect of Herba Lysimachiae extract (30 μg / ml) on glucose uptake in myocytes. C2C12 cells were inoculated and grown to confluence (3 days). Cells were differentiated for 4 days to form myotubes and then treated with a 24-hour treatment with a holly extract. Following supplementation, glucose was depleted for 24 hours, followed by serum depletion overnight. After washing, the fluorescent glucose analog 2-NBDG (2-(N-(7-nitrobenzo-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose) was added to the culture medium to monitor glucose uptake into living cells. After 30 minutes, fluorescence was measured after washing and cell lysis. n = 6 cells were assayed.

[0125] The results showed that at 25 μg / ml, the extract of Herba Lycopodii increased myocyte proliferation ( Figure 9 ), at 30 μg / ml, the extract of Oleander increased glucose uptake ( Figure 10 ).

[0126] Example 6

[0127] This example demonstrates the ability of an extract of Oleander to support liver detoxification function following aflatoxin B1 intoxication. Aflatoxin B1 was used as a model of liver intoxication.

[0128] Extracts of Olea europaea have been used in hepatoprotective studies. The human hepatocellular carcinoma cell line HepG2 is widely accepted as a model system for evaluating the toxic effects of various substances because these cells express most of the metabolic enzymes found in hepatocytes that catalyze biotransformation reactions. Therefore, this cell line is suitable for testing compounds for their potential hepatoprotective effects.

[0129] HepG2 cells were cultured at 1.6×10 4 Cells were seeded at a density of 10 cells / 0.2 mL in a 96-well plate and cultured for 24 hours in 90% Dulbecco's modified Eagle's medium (DMEM) containing 10% heat-inactivated fetal bovine serum (FBS). The cells were then supplemented with three doses (125, 250, and 500 μg / mL) of Herba Lycopersici extract and 1.5 μM aflatoxin B1. Cell viability was determined 48 hours later using a resazurin assay. The assay was repeated three times with three concentrations of Herba Lycopersici extract. A control was performed without aflatoxin B1 and Herba Lycopersici extract.

[0130] The results showed that even the lowest dose of the solstice extract reduced the toxicity of aflatoxin B1 ( Figure 11 ). This suggests that hollyhock may be used as a liver support compound, ultimately improving the condition and performance of farm animals.

[0131] Example 7

[0132] In this example, the effects of a Herba Oleraceae extract on markers of osteoarthritis were evaluated in an in vitro cell-based assay.

[0133] Chondrocytes were seeded in 96-well plates and cultured in culture medium for 24 hours. The culture medium consisted of Dulbecco's modified Eagle's medium supplemented with 4 mM L-glutamine, 50 U / mL penicillin and 50 μg / mL streptomycin, and 10% fetal bovine serum. The culture medium was then removed and replaced with assay medium containing or without (control) test compounds or reference substances (0.1 μM dexamethasone for MMP-3 and 1 μM indomethacin for PGE2), and the cells were preincubated for 24 hours. The assay medium consisted of Dulbecco's modified Eagle's medium supplemented with 4 mM L-glutamine, 50 U / mL penicillin and 50 μg / mL streptomycin, and 2% fetal bovine serum. After preincubation, the culture medium was removed and replaced with assay medium containing or without (control) test compounds or reference substances in the presence of a cytokine cocktail (IL-1β+TNF-α+IFN-γ+IL-6, 3 ng / ml each), and the cells were incubated for 48 hours. Non-stimulated controls were performed in parallel. All experimental conditions were performed with n=3.

[0134] After the incubation period, the supernatant was collected and the release of the inflammatory marker prostaglandin E2 (PGE2) was quantitatively analyzed. The levels of matrix degrading enzymes (MMP-3, a metalloproteinase involved in cartilage degradation) were determined using specific ELISA kits according to the supplier's instructions. The assay kits were Enzo Life Sciences, Ref. ADI-901-001 (for PGE2) and R&D Systems, Ref. DY513 (for MMP-3).

[0135] The results are as follows. 300 μg / mL of Herba Sophorae flavescentis extract reduced PGE2 production ( Figure 12 ), 300 μg / mL of Herba Sophorae flavescentis extract reduced matrix metalloproteinase-3 ( Figure 13 ).

[0136] Example 8

[0137] This example illustrates the positive effects of including the aerial parts of solstice grass in the diet on the performance of farm animals.

[0138] This study evaluated the effects of feeding a single dose of hollyhock to broiler chickens from 1 to 14 days of age on growth performance, feed intake, and feed efficiency (measured by feed conversion ratio). Treatments included two control diets and one experimental diet.

[0139] The basal diet composition was as follows (expressed in kg / 100 kg): corn 35.00, oats 14.00, barley 7.00, vegetable oil 6.00, corn gluten meal 1.00, soybean meal 29.00, oilseed meal 3.00, monocalcium phosphate 1.80, limestone 0.40, sodium chloride 0.40, lysine 0.50, methionine 0.60, and a premix containing other minerals and vitamins 0.500. The nutritional composition of diet 1 was 12.878 MJ / kg, with crude protein 22.094%, crude fat 8.681%, fiber 4.203%, lysine 1.559%, and methionine 0.7907.

[0140] Treatment groups were obtained by adding compounds to the basal diet. Diet 1 served as a negative control and did not include any compound. Diet 2 served as a positive control diet; it was supplemented with 100 g of Evolution-B, Code X60-6930, is approved as an alternative to antibiotic growth promoters (EFSA Journal, 2015). Diet 3: 157g of summer solstice grass per 1000kg feed.

[0141] One-day-old male Ross 308 chicks were obtained from a commercial hatchery. Upon arrival, 480 chicks were distributed into 24 floor pens with 20 chicks per pen. Each pen had a floor area of ​​150 × 140 cm.2 The chickens were housed in a solid floor covered with litter (wood shavings). The chickens were fed one of three experimental diets. Throughout the experiment, the chickens had ad libitum access to feed and water.

[0142] Three treatment groups (diet 1-3, described above) were fed to eight birds in separate pens using a randomized block design. Growth and feed intake information was collected from one to 14 days of age. Room temperature was approximately 32°C at one day of age and gradually lowered to 20°C by the end of the 21-day feeding period. A standard lighting schedule for broilers was used. Birds were weighed at the start (one day of age) and at 14 days of age, and feed efficiency was determined.

[0143] The results are summarized in Figure 14 Feed efficiency was estimated using feed conversion ratio. The main results are summarized below. Evolution-B, Code X60-6930, improved feed efficiency compared to the control group. The addition of solstice grass also improved the production performance of chickens, as feed efficiency increased by 5.1% compared to the control group.

[0144] Example 9

[0145] In this example, different plant parts (heartwood, bark, sapwood, branches with a diameter of 2.5 cm and leaves) of the same plant (Guaiacaceae) were evaluated for their total lignan, furanoguaiac and guaiac content.

[0146] Plant samples were ground into a fine powder and extracted with 80% v / v ethanol in an ultrasonic bath for 2 × 15 min, using a drug:solvent ratio of 1:50. The crude extract was centrifuged at 14,500 rpm for 5 min, and the supernatant was directly used for high-performance liquid chromatography (HPLC) analysis.

[0147] The HPLC system is equipped with an automatic sample injector and a diode array detector. A chromatographic column (e.g., Zorbax SB-C18 4.6*250mm, 5 μm) filled with octadecylsilyl silica gel is used as the stationary phase. The mobile phase is composed of solvent A (5% acetic acid) and solvent B (acetonitrile / methanol 50 / 50 (v / v)). Linear gradient elution is carried out at a flow rate of 1.5 ml / min. First, the detection of the lignan peak is carried out at 220, 280 and 320 nm. To measure the total lignan content, all lignan peaks in the chromatogram are identified by their UV spectra, which consist of two or three maxima at 238, 280 and at 230, 280 and 320 nm. For quantitative analysis, nordihydroguaiaretic acid (NDGA) is used as an external standard to calculate the sum of the lignan peak areas, because the UV spectrum of NDGA is similar to that of guaiac. NDGA (Sigma Aldrich, art. No. 74540), guaiacine (ambinter, c / o Greenpharma; No. amb35820270), and vanillin (Sigma Aldrich, No. V1104) were used as reference compounds. The UV spectra of all relevant peaks in the HPLC chromatogram were determined. The obtained UV spectra were then compared with known UV spectra (i.e., spectra published in the literature or spectra obtained with reference compounds). Based on the similarity of the spectra, peaks were identified or at least assigned to specific substance types. To verify the UV spectral similarity, the spectra of the peaks were overlapped with the spectra of the reference compounds (vanillin, guaiacine). The degree of similarity was then calculated using HPLC software. Lignans were quantified using NDGA as a reference compound.

[0148] The results showed the composition of lignan components in different plant parts of Guaiac wood. For example, Figure 15 and Figure 16 HPLC chromatograms showing the lignan components in heartwood and branches, respectively. Figure 17 The total lignan concentrations of the sacred guaiac plant parts are shown. Compared to the heartwood, the concentrations in the branches are 2-3 times lower, but still higher than in other parts of the guaiac (sapwood, bark); no relevant lignan content was observed in the leaves. Finally, Figure 18 The composition of furanoguaiacins, guaiacins, and guaiaretic acids in the total lignan fraction is shown in Table 1. Surprisingly, the lignan profiles in the branches were very similar to those in the heartwood.

[0149] Example 10

[0150] This example is used to illustrate the effect of total lignans standardized with furanoguaiacins and guaiacins on the production performance of farm animals.

[0151] The study evaluated the effects of supplementing a diet standardized for total lignans, including furanoguaiacoids and guaiacoids, on growth performance, feed intake, and feed efficiency (measured by feed conversion ratio) when fed to broiler chickens aged 1 to 28 days. Treatment groups included two control diets and one experimental diet.

[0152] The basal diet composition was as follows (expressed in kg / 100 kg): corn 35.00, oats 14.00, barley 7.00, vegetable oil 6.00, corn gluten meal 1.00, soy flour 29.00, rapeseed meal 3.00, monocalcium phosphate 1.80, limestone 0.40, NaCl 0.40, lysine 0.50, methionine 0.60, and a premix containing other minerals and vitamins 0.500. The nutritional composition of diet 1 was 12.878 MJ / kg, crude protein 22.094%, crude fat 8.681%, fiber 4.203%, lysine 1.559%, and methionine 0.7907%.

[0153] Treatment groups were obtained by adding compounds to the basal diet. Diet 1 served as a negative control and did not include any compound. Diet 2 served as a positive control diet; it was supplemented with 100 g of Evolution-B, Code X60-6930, is approved as an alternative to antibiotic growth promoters (EFSA Journal, 2015). Diet 3 was supplemented with 8g of total lignans per metric ton of diet 1, consisting of 50% furanoguaiac compounds and 40% guaiac compounds.

[0154] Approximately 400 one-day-old male Ross 308 chicks were obtained from a commercial hatchery. Upon arrival, the 400 chicks were divided into 20 floor pens with 20 chicks per pen. The floor area of ​​each pen was 150 × 140 cm. 2 The chickens were housed in a solid floor covered with litter (wood shavings). The chickens were fed one of three experimental diets. Throughout the experiment, the chickens had ad libitum access to feed and water.

[0155] Three treatment groups (diet 1-3 described above) were fed to eight chickens in separate pens using a randomized block design. Growth and feed intake information was collected from one to 28 days of age. Room temperature was approximately 32°C at one day of age and gradually decreased to 20°C by the end of the 21-day feeding period. A standard lighting program for broilers was used. Birds were weighed at the beginning (one day of age), 14 days of age, and at the end (28 days of age) of the study, and weight gain and feed efficiency were determined.

[0156] The results are summarized in Figure 19 The feed efficiency was estimated by feed conversion ratio. The main results are summarized as follows: 1) 100g per ton of feed was added Evolution-B, Code X60-6930 improves feed efficiency to a similar extent as the published information for this product. For example, Bravo et al. (J. Appl. Poul. Res. 20: 115-120, 2009) showed that adding the same product at the same dose to chickens increased feed efficiency by 2.28% (feed conversion ratio: 34 days of age, 1.75 vs. 1.79). In this study, the same product increased feed efficiency by 2.2% (feed conversion ratio: 28 days of age, 1.60 vs. 1.57); and 2) the addition of 8 ppm of total lignans composed of 50% furanoguaiac compounds and 40% guaiac compounds significantly improved the production performance of chickens. Feed efficiency increased by 4.7% (feed conversion ratio: 1.52 vs. 1.60).

[0157] Example 11

[0158] In this example, the effect of increasing the dose of total lignans standardized with furanguaiac compounds and guaiac compounds on in vitro anti-inflammatory cytokines is described. The inhibitory effect of 70% v / v ethanol guaiac extract standardized with total lignans, furanguaiac and guaiac on cytokine release was determined using THP-1 human monocyte cell line differentiated into macrophage-like cells by phorbol myristate (PMA). Prior to the assay, mitochondrial dehydrogenase activity in living cells was determined using MTT (bromide 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium) assay to determine the potential cytotoxic effects of the test compound. The anti-inflammatory effect on cytokine release in THP-1 macrophages was measured by incubating the cells with three concentrations of guaiac extract and subsequently adding LPS to stimulate cytokine production. After 24 hours, the amount of TNF-α, IL-1β and IL-6 in the cell culture supernatant was determined by capture ELISA using a commercially available kit.

[0159] The main results were: 1) Increasing doses of total lignans standardized to furanoguaiacins and guaiacins reduced the production of the inflammatory cytokine TNFα (see Figure 20 ); 2) increasing the dose of total lignans standardized with furanoguaiacins and guaiacins reduced the production of the inflammatory cytokine IL6 (see Figure 21 ); and 3) increasing the dose of total lignans standardized to furanoguaiacins and guaiacins reduced the production of the inflammatory cytokine IL1β (see Figure 22 ).

[0160] Example 12

[0161] In this example, the effects of increasing doses of total lignans, normalized to furanoguaiacins and guaiacins, on osteoarthritis markers were evaluated in an in vitro cell-based assay.

[0162] Chondrocytes were seeded in 96-well plates and cultured in culture medium for 24 hours. The culture medium consisted of Dulbecco's modified Eagle's medium supplemented with 4 mM L-glutamine, 50 U / mL penicillin and 50 μg / mL streptomycin, and 10% fetal bovine serum. The culture medium was then removed and replaced with assay medium containing or without (control) test compounds or reference substances (0.1 μM dexamethasone for MMP-3 and 1 μM indomethacin for PGE2), and the cells were pre-incubated for 24 hours. The assay medium consisted of Dulbecco's modified Eagle's medium supplemented with 4 mM L-glutamine, 50 U / mL penicillin and 50 μg / mL streptomycin, and 2% fetal bovine serum. After pre-incubation, the culture medium was removed and replaced with assay medium containing or without (control) test compounds or reference substances in the presence of a cytokine cocktail (IL-1β+TNF-α+IFN-γ+IL-6, 3 ng / ml each), and the cells were incubated for 48 hours. Non-stimulated controls were performed in parallel. All experimental conditions were performed with n=3.

[0163] At the end of the incubation period, the supernatant was collected and the release of the inflammatory marker prostaglandin E2 (PGE2) was quantitatively analyzed, and the level of matrix degrading enzymes (MMP-3, a metalloproteinase involved in cartilage degradation) was determined using specific ELISA kits according to the supplier's instructions. The assay kits were Enzo Life Sciences, Ref. ADI-901-001 (for PGE2) and R&D Systems, Ref. DY513 (for MMP-3).

[0164] The results are as follows. Increasing doses of total lignans (0.5, 1, and 5 μg / mL) standardized with furanoguaiacins and guaiacins reduced PGE2 production ( Figure 23 ), and reduced matrix metalloproteinase-3 ( Figure 24 ).

[0165] Example 13

[0166] This study evaluated the effects of supplementing guaiac wood and holly extracts (70 ppm each, defined as PP2) with broiler chickens aged 1 to 42 days on growth performance, feed intake, and feed conversion. Treatments included a control diet and four color-coded treatment diets. The control diet contained no additives; experimental diet 1 contained supplement D; experimental diet 2 contained PP2; experimental diet 3 contained PP2 plus supplement D; and experimental diet 4 contained PP2 plus eugenol. The study feeding schedule included different phases: a starter diet for 0 to 14 days; a grower diet for 14 to 28 days; and a finisher diet for 28 to 42 days.

[0167] Table 2

[0168]

[0169]

[0170] Treatment groups were obtained by supplementing the basal diet with one or a combination of compounds. Diet 1 served as a negative control and did not include any compounds. Diet 2 served as a positive control diet; it included 100 g / metric ton of Supplement D, a product that has been developed as a growth promoter. Diet 3 was a combination of Diet 1 supplemented with 70 g each of Guaiac and Oleander extracts. Diet 4 was Diet 1 supplemented with 70 g each of Guaiac and Oleander extracts and 100 g / metric ton of Supplement D. Diet 5 was Diet 2 supplemented with 10 g / metric ton of eugenol.

[0171] Test methods

[0172] Chicken numbers, allocation, pens, feed, water, lighting and vaccinations

[0173] A total of 1860 newly hatched (d 0) Ross 708 male chickens were obtained from a commercial hatchery. After day 7, 360 chickens were assigned to each treatment, with 12 replicates per treatment and 31 chickens per replicate per treatment.

[0174] distribute

[0175] The birds were randomly assigned to pens and grouped throughout the shed. Each block was represented by a different colored pens, with the treatment group listed in a different color next to the pens. Each treatment group had 12 replicate pens. All pens had 31 birds each and were placed on day 0 of the experiment. On day 7, all pens were re-counted to 30 birds per pen. If more than one bird was lost per pen, it was necessary to make up the loss from the extra birds fed to the same treatment group on days 1-7.

[0176] Stalls, feed and water

[0177] All pens in this study were lined with dirt litter (from the previous flock - all litter was removed from the pens, mixed, and then redistributed to the pens in this study). The starter diet (0-14 days) was a mash diet, and the remaining diets were litter, which was provided by the University of Illinois Feed Mill and Kalmbach Feed Inc. All treatment groups had free access to water.

[0178] illumination

[0179] From days 0 to 8, the birds were exposed to 24 hours of full-intensity light. From days 9 to 25, the lighting program was changed to provide 16 hours of light and 8 hours of darkness, and from days 24 to 42, 18 hours of light were provided.

[0180] Vaccination

[0181] All chickens are vaccinated against Marek's disease and infectious bronchitis / Newcastle disease at the hatchery. In addition, chickens are vaccinated against coccidiosis at the hatchery.

[0182] Using a randomized block design, five treatment groups (diet 1-5 described above) were fed to 60 birds in separate pens. Growth and feed intake information was collected from days 0 to 42 of age. The following data were collected: 1. Pen weights at days 0, 7, 14, 21, 28, and 42; 2. Feed addition, consumption, and removal at each stage; 3. Average daily gain, feed intake, and feed conversion ratio for each pen from days 0-14, 0-28, and 0 / 42; and 4. Mortality, cause of death, and removal.

[0183] result

[0184] Initial stage

[0185] As previously described, all birds in all treatment groups were fed a mash starter diet. Weight gain, feed intake, and feed conversion ratio (FCR) were calculated as described above. The FCR was adjusted for mortality. Data obtained during the starter period are shown below.

[0186] Table 3

[0187]

[0188] The results of the average weight of chickens from 0 to 14 days are as follows Figure 25 The results of FCR adjusted by the number of deaths on days 0-14 are shown in Figure 26As shown in the results, no clear numerical trends were observed for the mean body weight and mortality-adjusted FCR results from days 0 to 14. This is consistent with the fact that phytogenic molecules require 3-4 weeks to exert their effects on physiological responses and, consequently, on zootechnical parameters. A possible synergistic effect between PP2 and Supplement D on mortality-adjusted FCR was observed during the first two weeks of the trial.

[0189] Growth period

[0190] All chickens were fed with the corresponding feed in pellet form. Table 4 below shows the parameters of different growth stages as follows.

[0191] Table 4

[0192]

[0193] The results of the average weight from 0 to 28 days are as follows Figure 27 As shown, the FCR results adjusted for the number of deaths on days 0-28 are as follows Figure 28 As shown. As can be seen from the results, all experimental diets (2, 3 and 4) had a positive effect on the mortality-adjusted FCR at the end of day 28. When compared to the control diet, experimental diet 1 with supplement D showed a 1.78% improvement. Experimental diets 2, 3 and 4 showed increases in mortality-adjusted FCR of 4.8%, 3.53% and 2.14%, respectively. The response of treatment 3 is very important because it showed the same response as reported in the previous trial, with a percentage improvement of 4.2 in FCR.

[0194] Fattening stage

[0195] A summary of the data for days 0-42 is shown in Table 5 below.

[0196] Table 5

[0197]

[0198] The results of the average weight from 0 to 42 days are shown in Figure 29 The results of FCR adjusted for the number of deaths on days 0-42 are shown in Figure 30 At day 42, all experimental diets showed an improvement in feed conversion ratio when compared to the control group. Experimental diet 2, supplemented with PP2, showed a significant improvement in FCR at day 42. Experimental diet 2 showed a 2.4% improvement compared to the control group. The combination of PP2 with supplement D (experimental diet 3) and eugenol (experimental diet 4) also showed an improvement in FCR. This data suggests that PP2, alone and / or in combination with other compounds such as carvacrol, cinnamaldehyde, and capsaicin (components of supplement D) and eugenol, has a positive impact on zootechnical parameters in chickens.

[0199] Supplement D, at 136g per metric ton, showed a significant increase in FCR. In this study, the same product improved feed efficiency by nearly 2.0% (feed conversion ratio: 1.19 vs. 1.17 at 28 days of age). Adding extracts of Guaiac Wood and Oleander (70ppm each, defined as PP2) increased FCR by 4.8% and 2.7%, respectively, significantly improving performance at days 28 and 42. Extracts of Guaiac Wood and Oleander have positive effects on chicken performance, both alone and in combination with other compounds.

[0200] Example 14

[0201] The effects of feeding different feed products on broiler performance variables from day 1 to 42 were evaluated. The objective of this study was to determine the performance parameters of live weight gain, feed conversion, and mortality in broiler chickens raised in floor pens that were vaccinated against coccidiosis and treated with different products. The experiment consisted of 78 pens of 50 male broiler chickens each. Treatments were replicated in 13 blocks; six treatments were randomly assigned within each block. Pen assignment to treatment and block was randomized. Pen maps and treatment assignments are included in the source data. Treatments are shown in Table 6 below.

[0202] Table 6

[0203]

[0204] diet

[0205] Southern Poultry Research, Inc. provided all feeds. All basal feeds were produced at the SPFR feed facility. Feed formulations are provided in the source data. Test samples were used according to the procedures described in the "Treatment" section. All feeds were fed in the form of crumbles / pellets. The quantities of all basal feeds and test samples used to prepare treatment batches were recorded. All test samples were mixed to ensure uniform distribution. Each batch of feed was mixed and bagged individually. Each bag was labeled with the study number, mixing date, feed type, and the correct treatment number. Complete records of feed mixing and test sample inventory were maintained.

[0206] Feed samples

[0207] Samples were collected from the beginning, middle, and end of each treatment feed and combined to form a composite sample. Each treatment was sampled from the composite sample. Each treatment was sampled once.

[0208] Feeding plan

[0209] Feed will be weighed per pen. Starters will be fed from DOT 0-14. At DOT 14, any uneaten starters will be weighed and discarded. Finishers will be released and fed until DOT 28. At DOT 28, any uneaten growers will be weighed and discarded. Finishers will be released and fed until DOT 42. At DOT 42, any uneaten finishers will be weighed and discarded.

[0210] chicken

[0211] Day-of-hatch male chickens were obtained from Cobbhatchery, Cleveland, GA. The breed was Cobb 500. A record of the breeder flock was kept. 3,000 birds were allocated for the study. At the hatchery, the birds received routine vaccinations. The birds were sexed at the hatchery. Only healthy-looking birds were used in the study. At the start of the study, 50 male birds were allocated to each treatment pen in blocks. No birds were replaced during the study. The disposition of all birds not allocated was recorded. Bird weights were recorded on days 0, 14, 28, and 42.

[0212] Ground Fence Description and Management

[0213] Floor-pen chicken houses are modified chicken houses with dirt floors and curtain walls. The houses will be prepared for research use according to SPFR standard operating procedures. SPFR will include a schematic diagram of the test facility in the source data.

[0214] The experimental henhouse was divided into pens of equal size, arranged along a central aisle. Chickens were housed in 60 pens in a house equipped with 80 floor pens, each measuring 5 x 10 = 50 square feet. The bedding consisted of wood shavings approximately 4 inches thick. After deducting equipment, the stocking density was approximately 0.95 square feet per bird. Each pen had 5-foot-high sidewalls and a solid wood bottom 1.5 feet to prevent migration. All pens were consecutively numbered and identified on pen cards.

[0215] The building temperature will be monitored. Environmental conditions (temperature) during the experiment will be appropriate (optimal) for the age of the animals. Light will be provided by fluorescent bulbs placed above the pens. The lighting regimen will be 24 hours per day.

[0216] Each pen had free access to feed via a tube feeder. From day 0 to day 7, feed was also provided in a tray placed on the bedding in each pen. Each pen had free access to water via a Plasson-type automatic drinker.

[0217] Standard floor-level pen management practices were used throughout the experiment. Animals and housing facilities were inspected twice daily to observe and record overall health, consistent feed and water availability, and temperature. All birds that died were removed and identified as incidental. Birds found dead during the study were recorded in the daily mortality log and were not replaced. Pen number, date of death, sex, weight, and diagnosis were recorded.

[0218] Data entry and analysis

[0219] Source data should be entered in indelible ink. Records must be legible, signed or initialized by the person observing the record, and dated. Each source data sheet will be signed by the person to whom the data relates. Any errors or changes to the source data will be initialized and dated, and a correction code or statement will be added to explain the reason for the change. Average pen weight gain, feed consumption, and feed conversion ratio will be calculated for each feeding period (Days 0-14, 0-35, and 0-42).

[0220] Example 15

[0221] Effects of feed plants 1 and 2 on broiler performance. Total number: 480; Experimental duration: 21 days; Genetics: ROSS308FF x Yield Plus; Treatment number: 4; Quantity: 4; Replication number: 12; Number of birds / pen: 10 / cage.

[0222] Chickens had free access to feed and water. Routine health procedures were followed. The study director was notified of any therapeutic treatments. A veterinarian was consulted for any treatments not standard for the unit. Challenge Model: A commercial coccidiosis vaccine was administered orally by gavage 10X upon placement in the cage, as recommended by the manufacturer. This oral gavage consisted of 10X coccidiosis vaccine and distilled water, totaling 1 mL. A wooden board was placed near the feeder / drinker in each cage to allow for natural Eimeria circulation. A total of four diets were fed according to the following feeding schedule for quantitative use (and treatment identification) (Table 7).

[0223] Table 7

[0224]

[0225] The mixed basal diets were manufactured by Deal-Rite Feeds (2000 lb minimum batches). The experimental feeds were produced in a cement mixer at the experimental site. 400 lb of feed was produced for each treatment. All diets were fed ad libitum. Observations: Pen weights and feed consumption were recorded on days 0, 7, 14, and 21; any abnormal observations were recorded; any mortalities or removals were weighed and recorded; all health events and subsequent dispositions (bird ID, observed health issues) were recorded. Reports: Raw data - animal weights, feed consumption; laboratory analyses; data summaries. Animal housing and use were the same as those conventionally maintained in commercial poultry production. Samples: Day 21 intestinal samples of 5-10 birds / trt were collected - using the same blocks as in the previous experiment; two samples of each diet were collected. Two 1-lb samples were collected for each diet. One sample of each diet was sent to Sure-Tech labs for testing of moisture, crude protein, fat, Ca, P, Cu, Na, Zn, chloride, Mg, and Mn. A subsample was retained.

[0226] Data summary and statistical analysis

[0227] Table 8

[0228]

[0229] The data will be analyzed using a completely randomized design with the pen as the experimental unit and block-based on initial body weight. Data analysis was performed using SAS mixed program. The results for day 21 are shown in Table 9 (FCR is feed conversion ratio).

[0230] Table 9

[0231]

[0232] To facilitate understanding of this disclosure, a number of terms are defined below. The terms defined herein have the meanings that are commonly understood by a person of ordinary skill in the art to which this disclosure relates. In the claims and / or in the specification. The words "a" or "an" when used in conjunction with the term "comprising" can mean "one", but they are also consistent with the meanings of "one or more", "at least one" and "one or more than one". The term "or" used in the claims means "and / or" unless it is expressly indicated that only alternatives are referred to or the alternatives are mutually exclusive, although this disclosure supports definitions that refer only to alternatives and "and / or". In this application, the term "about" is used to indicate that a value includes the inherent error variation of the device, the error variation of the method used to determine the value, or the error variation that exists in the subject of study.

[0233] As used in this specification and claims, the words "comprising" (and any form of comprising, such as "containing" and "including"), "having" (and any form of having, such as "having" and "having"), "including" (and any form of including, such as "containing" and "including"), or "containing" (and any form of containing, such as "including" and "including") are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. In any of the embodiments of the compositions and methods provided herein, "comprising" may be replaced with "consisting essentially of" or "consisting of." As used herein, the phrase "consisting essentially of requires the specified integers or steps, and these integers or steps do not materially affect the characteristics or functions of the claimed invention. As used herein, the term "consisting of" is used only to indicate the presence of a recited integer (e.g., a feature, element, characteristic, property, method / process step, or limitation) or combination of integers (e.g., a feature, element, characteristic, property, method / process step, or limitation).

[0234] As used herein, the term "or combinations thereof" refers to all permutations and combinations of the items preceding the term. For example, "A, B, C, or combinations thereof" is intended to include at least one of: A, B, C, AB, AC, BC, or ABC, and if order is important in a particular context, BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing with this example, specifically included are combinations containing repetitions of one or more items or terms, such as BB, AAA, AB, BBC, AAABCCCC, CBBAAA, CABABB, etc. Those skilled in the art will understand that, unless the context clearly dictates otherwise, there is generally no limit to the number of items or terms in any combination.

[0235] As used herein, approximating words such as, but not limited to, "about," "substantially," or "substantially" mean that when so modified, it should be understood that the value is not necessarily absolute or perfect, but should be considered close enough to indicate that such a situation exists for a person skilled in the art. The degree to which the description can vary depends on how much change can be made and still recognize that the modified feature still has the desired properties and functions of the unmodified feature. Generally, but in accordance with the foregoing discussion, a numerical value modified by an approximating word such as "about" herein can vary from the stated value by at least ±1, 2, 3, 4, 5, 6, 7, 10, 12, or 15%.

[0236] According to the present disclosure, all compositions and / or methods disclosed and claimed herein can be prepared and implemented without excessive experimentation. Although the compositions and methods of the present disclosure have been described according to preferred embodiments, it will be apparent to those skilled in the art that, without departing from the concept, spirit and scope of the present disclosure, the steps or sequence of steps of the compositions and / or methods and methods described herein may be changed. For example, all disclosed components of the preferred embodiments and alternative embodiments are interchangeable, thereby providing a variety of systems comprising a combination of all preferred embodiments and alternative embodiment parts. Therefore, the present application is intended to cover any changes, uses or modifications of the present disclosure using its general principles. In addition, the present application is intended to encompass such deviations from the present disclosure within the known or conventional practices of the field to which the present disclosure pertains and that fall within the limitations of the appended claims. All such similar substitutions and modifications apparent to those skilled in the art are considered to be within the spirit, scope and concept of the present disclosure as defined by the appended claims.

Claims

1. An animal feed ingredient composition comprising an effective amount of Ribes alpina or an extract thereof, wherein the effective amount improves the performance of an animal feed to which the ingredient is added; and wherein the composition further comprises: a) an effective amount of furanoguaiac or a furanoguaiac-like compound or guaiac or a guaiac-like compound, or a biologically acceptable salt thereof, wherein said effective amount improves the performance of an animal feed to which said ingredient is added; b) an effective amount of aestivin, or a biologically acceptable salt thereof, wherein said effective amount improves the performance of an animal feed to which said ingredient is added; or c) effective amounts of a) and b).

2. The composition of claim 1, further defined as comprising at least one animal feed component selected from the group consisting of vitamins, minerals, probiotics, enzymes, flavorings, amino acids, fats, essential oils, and preservatives.

3. The composition of claim 1, wherein the extract is produced from the stem, leaves, flowers, roots or fruits of Ribes alpina or parts thereof, or any combination thereof.

4. The composition of claim 1, wherein the extract is an extract of water, ethanol, methanol, isopropanol, ethyl acetate, acetone, hexane or a mixture thereof, or an extract of supercritical CO2.

5. The composition of claim 1, wherein the Ribes alpina or extract thereof comprises from about 3 g / metric ton to about 500 g / metric ton of the composition.

6. The composition of claim 5, wherein the Ribes alpina or extract thereof comprises from about 9 g / metric ton to about 90 g / metric ton of the composition.

7. The composition of claim 1, wherein the effective amount of a), b) or c) is present in an amount effective to improve the performance of the animal feed to which the ingredient is added relative to when the effective amount of a), b) or c) is not present in the animal feed.

8. The composition of claim 1, wherein: furanoguaiacine or a furanoguaiac-like compound or guaiacine or a guaiac-like compound obtained from Machilus edulis, Persea fructifera, Machilus thunberghii, Cinnamomum philippense, Saurus cernuus, Myristica fragrans, Guaiacum sanctum, or Guaiacum officinale plants or plant parts, or extracts thereof; or The aestivin is obtained from Marrubium vulgare, Marrubium velatinum, Marrubium cylleneum, Marrubium trachyticum, Marrubium globosum, Marrubium anisodon, Marrubium sericeum, Marrubium supinum, Leonotis leonurus, Leonotis nepetifolia, Phlomis bracteosa, Marrubium deserti de Noe, Marrubium alysson, or Marrubium thessalum plants or plant parts, extracts thereof, or any combination thereof.

9. The composition of claim 8, wherein: The furanoguaiac or furanoguaiac-like compound or guaiac or guaiac-like compound is obtained from the heartwood or branches of the sacred guaiac tree or the guaiac plant, or an extract thereof; or The aestivin is obtained from the aerial parts of the Aesculus serrata plant, or an extract thereof.

10. The composition of claim 1, wherein the furanoguaiacine, furanoguaiac-like compounds, guaiacine, guaiac-like compounds, and guaiacine are synthetically prepared.

11. The composition of claim 1, wherein the marrubiin is converted into marrubiinic acid, marrubenol, 12-S-hydroxymarrubiin, 11-oxo-marrubiin, premarrubiin, 3-deoxo-15-S-methocyvelutein, marrilibalacetal, cyllenin A, polydinonine, preleosibirin, or peregrinol.

12. The composition of claim 1, wherein: Furanoguaiacin compounds or guaiacin compounds have the following chemical structure: Wherein R1 is -OH, -OCH3 or -OCH2, R2 is -OCH3 or -O, R3 is -OCH2 or -O, and R4 is -O or OCH3, and the corresponding glycoside form, the summer grass has the following chemical structure: or The summer solstice grass has the following chemical structure:

13. The composition of claim 1, wherein the lignans comprise from about 0.5% to about 30% of the composition.

14. The composition of claim 1, wherein lignans comprise at least 9% of the composition.

15. The composition of claim 1, wherein the total lignans in the composition comprise at least 20% furanoguaiacine or furanoguaiac-like compounds and guaiacine or guaiac-like compounds.

16. The composition of claim 1, further wherein the guaiacine or guaiac-like compound comprises about 20-60 weight % of the total lignans contained in the composition.

17. The composition of claim 1, further wherein the furanoguaiac or furanoguaiac-like compound comprises about 20-60 weight % of the total lignans contained in the composition.

18. An animal feed comprising the animal feed ingredient composition of claims 1-17 in an amount effective to improve the zootechnical performance of an animal fed the animal feed relative to a control feed lacking the animal feed ingredient composition.

19. The animal feed of claim 18, wherein the animal feed is chicken feed, pig feed, dairy cow feed, or beef cattle feed.

20. The animal feed of claim 18, wherein the improved zootechnical performance is improved muscle function or health, altered feed intake, increased average daily weight gain, improved feed efficiency, decreased feed conversion ratio, or increased milk production.

21. The animal feed of claim 18, wherein: The amount of Ribes alpina or its extract is about 3 g to about 500 g per metric ton of animal feed; or The amount of aestivum or a biologically acceptable salt thereof is about 0.5 mg to about 160 mg / kg of the animal feed.

22. A method for improving the performance of animal feed, comprising adding an effective amount of the composition of claim 1 to the animal feed.

23. The method of claim 22, wherein the improved performance comprises improved muscle function or health, altered feed intake, increased average daily weight gain, improved feed efficiency, decreased feed conversion ratio, or increased milk production in an animal fed the animal feed relative to a control animal feed to which the animal feed ingredient composition is not supplemented.

24. The method of claim 22, wherein the animal feed is chicken feed, pig feed, dairy cow feed, or beef cattle feed.

25. The method of claim 22, wherein: The amount of Ribes alpina or its extract is about 3 g to about 500 g per metric ton of animal feed; or The amount of summergrass or its biologically acceptable salt is about 0.5 mg to about 160 mg / kg of animal feed.

26. A method for improving the zootechnical performance of an animal comprising feeding said animal the animal feed of claim 18.

27. The method of claim 26, wherein the animal is a chicken, pig, dairy cow, or beef cattle.

28. The method of claim 26, wherein the improved zootechnical performance is improved muscle function or health, altered feed intake, increased average daily weight gain, improved feed efficiency, decreased feed conversion ratio, or increased milk production.

29. The method of claim 26, wherein: The animal feed comprises Ribes alpina or an extract thereof in an amount of about 3 g to about 500 g per metric ton of the animal feed; or The amount of furanoguaiac or furanoguaiac-like compound, or guaiac or guaiac-like compound or biologically acceptable salt thereof is about 0.01 mg to about 2000 mg / kg of animal feed.

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