Improving milk quality

Abstract

This disclosure relates to a method for improving milk quality in lactating mammals, comprising administering a composition comprising at least one casein-derived peptide.

Description

Technical Field

[0001] This application includes a sequence listing submitted electronically in ASCII format, which is hereby incorporated by reference in its entirety. The above ASCII copy was created on May 16, 2024, named P-625310-PC-ST26, and is 69,632 bytes in size.

Background Art

[0002] Casein proteins are composed of three fractions, α, β, and κ, according to their electrophoretic mobility. Casein hydrolysates are the hydrolyzed forms of casein, which contain, in particular, active β-casein-derived peptides. It has been established that casein hydrolysates play a role in the immune response against microbial and viral infections.

[0003] The global dairy product market is estimated to exceed $500 billion and has an average annual growth rate of 4%. Cow's milk occupies a significant portion of the market, while plant-derived alternatives account for $1 billion in the United States and are estimated to be $700 million for lactose-intolerant milk. Mammalian or mammalian-produced milk is a very complex fluid containing thousands of components. Mammalian or mammalian-produced milk contains water, various different lipids, sugars, various different proteins, and various different inorganic salts and compounds.

[0004] Dietary proteins are essential nutrients for human health and growth. The World Health Organization recommends that dietary proteins should contribute approximately 10 - 15% of the energy intake when the energy balance is in equilibrium and the body weight is stable. The average daily protein intake in each country indicates that these recommendations are consistent with the amount of protein consumed worldwide. A diet with an average of 20 - 3% of energy from protein is representative of a high-protein diet when consumed with an energy balance.

[0005] The body cannot synthesize certain amino acids necessary for health and growth and must obtain them from food instead. These amino acids are called “essential amino acids” and include histidine (H), isoleucine (I), leucine (L), lysine (K), methionine (M), phenylalanine (F), threonine (T), tryptophan (W), and valine (V). Dietary proteins that supply all essential amino acids are called “high-quality” proteins. Animal foods such as meat, fish, poultry, eggs, and dairy products are generally considered high-quality protein sources that provide a good balance of essential amino acids. Casein protein and whey (protein in the liquid remaining after milk has been curdled and strained) are major sources of high-quality dietary protein. Foods that do not supply a good balance of essential amino acids are called “low-quality” proteins. Most fruits and vegetables are poor sources of protein. Some plant foods, including beans, peas, lentils, nuts, and grains (e.g., wheat), are better sources of protein. Soybeans are a plant-based protein derived from soybeans and are sometimes considered a high-quality protein source.

[0006] The composition and characteristics of raw milk are crucial for controlling the quality of dairy products. There remains a great need for a source of high-quality nutritional products. [Overview of the Initiative]

[0007] In some embodiments, disclosed herein are methods for improving milk quality in lactating mammals, comprising administering a composition comprising at least one casein-derived peptide.

[0008] In some related embodiments, improved milk quality includes increased milk protein, increased milk fat, increased lactose, increased energy-corrected milk (ECM), or any combination thereof.

[0009] In some related embodiments, the proportion of milk protein in the milk is in the range of 2.5% to 6.5%. In some further related embodiments, the proportion of milk protein in the milk is increased by 0.5% to 30% compared to the proportion of milk protein in a control lactating mammal. Those skilled in the art will understand the phrase “control lactating mammal” as a mammal that has not been administered any treatment, or a mammal that has been administered a treatment different from the composition containing at least one casein-derived peptide.

[0010] In some relevant embodiments, the percentage of milk fat in milk is in the range of 2.5% to 6.5%. In some relevant embodiments, the percentage of milk fat in milk is increased by 0.5% to 30% compared to the percentage of milk fat in control lactating mammals.

[0011] In some relevant embodiments, the lactose content in milk is in the range of 3.5% to 7.8%. In some further relevant embodiments, the lactose content in milk is increased by 0.5% to 30% compared to the lactose content in control lactating mammals.

[0012] In some related embodiments, the extracorporeal membrane mass (ECM) is increased by 0.5% to 20% compared to the ECM in control lactating mammals.

[0013] In some embodiments, milk quality is improved at any stage of the lactation cycle.

[0014] In some embodiments, administration occurs at the start of the dry period. In some embodiments, milk quality is improved regardless of the length of the dry period.

[0015] In some related embodiments, the casein-derived peptides include natural peptides, synthetic peptides, semi-synthetic peptides, or any combination thereof.

[0016] In some further related embodiments, the casein-derived peptide comprises one or more fragments of β-casein, αS1-casein, αS2-casein, and κ-casein. In some further related embodiments, the casein-derived peptide further comprises amino acids having different lengths.

[0017] In some further related embodiments, the casein-derived peptide comprises a casein hydrolysate. In some further related embodiments, the casein-derived peptide comprises a phosphopeptide. In some further related embodiments, the phosphopeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 26.

[0018] In some embodiments, the composition is free of antimicrobial agents and includes an acceptable carrier.

[0019] In some embodiments, the method includes administering a casein-derived peptide in a concentration of 10 ng / ml to 500 mg / ml.

[0020] In some embodiments, administration involves intramammary injection into one or more nipples.

[0021] In some embodiments, milk is essentially free of residues and can be used as raw milk for dairy product manufacturing, lactation, bread making, confectionery making, feeding, or a combination thereof. In some further embodiments, dairy products include milk, whey, yogurt, cheese, cream, butter, high-protein milk beverages, or a combination thereof.

[0022] In some embodiments, disclosed herein are compositions comprising at least one casein-derived peptide for use in improving milk quality in lactating mammals.

[0023] The subject matter regarded as the present invention is particularly pointed out in the concluding part of the specification and is clearly claimed for a patent. The present invention, however, together with its objects, features, and advantages, can be best understood by reference to the following detailed description when read in conjunction with the accompanying drawings, with respect to both its construction and the method of operation.

Brief Description of the Drawings

[0024] [Figure 1] Diagram of daily / average energy-corrected milk (ECM) production (Kg) in dairy cows at the dry period stage treated with bovine casein hydrolysate (bCNH) over a subsequent lactation period of 305 days. Control - dairy cows were treated with an antimicrobial agent. bCNH11 - dairy cows were treated with a single dose of bovine casein hydrolysate (bCNH). bCNH22 - dairy cows were treated with two doses of bovine casein hydrolysate (bCNH). *p<0.05. Control: 36.67, bCNH11: 39.83, bCNH22: 39.62; [Figure 2] Diagram of monthly energy-corrected milk (ECM) production (Kg) in dairy cows at the dry period stage treated with bovine casein hydrolysate (bCNH) over a subsequent lactation period of 305 days. Control - dairy cows were treated with an antibiotic. bCNH11 - dairy cows were treated with a single dose of bovine casein hydrolysate (bCNH). bCNH22 - dairy cows were treated with two doses of bovine casein hydrolysate (bCNH). *p<0.05; [Figure 3] Diagram of daily / average milk fat yield (kg) in dairy cows at the dry period stage treated with bovine casein hydrolysate (bCNH) over a subsequent lactation period of 305 days. Control - dairy cows were treated with an antibiotic. bCNH11 - dairy cows were treated with a single dose of bovine casein hydrolysate (bCNH). bCNH22 - dairy cows were treated with two doses of bovine casein hydrolysate (bCNH). *p<0.05 bCNH11: 1.45, bCNH22: 1.49; [Figure 4]Graph of monthly milk fat yield (kg) in dairy cows at the dry period stage treated with bovine casein hydrolysate (bCNH) over a subsequent lactation period of 305 days. Control - dairy cows were treated with antibiotics. bCNH11 - dairy cows were treated with a single dose of bovine casein hydrolysate (bCNH). bCNH22 - dairy cows were treated with two doses of bovine casein hydrolysate (bCNH). *p<0.05; [Figure 5] Graph of daily / average milk protein yield (kg) in dairy cows at the dry period stage treated with bovine casein hydrolysate (bCNH) over a subsequent lactation period of 305 days. Control - dairy cows were treated with antibiotics. bCNH11 - dairy cows were treated with a single dose of bovine casein hydrolysate (bCNH). bCNH22 - dairy cows were treated with two doses of bovine casein hydrolysate (bCNH). *p<0.05. Control 1.23, bCNH11: 1.31, bCNH22: 1.36; [Figure 6] Graph of monthly milk protein yield (kg) in dairy cows at the dry period stage treated with bovine casein hydrolysate (bCNH) over a subsequent lactation period of 305 days. Control - dairy cows were treated with antibiotics. bCNH11 - dairy cows were treated with a single dose of bovine casein hydrolysate (bCNH). bCNH22 - dairy cows were treated with two doses of bovine casein hydrolysate (bCNH). *p<0.05; [Figure 7] Graph of daily / average milk lactose yield (kg) in dairy cows at the dry period stage treated with bovine casein hydrolysate (bCNH) over a subsequent lactation period of 305 days. Control - dairy cows were treated with antibiotics. bCNH11 - dairy cows were treated with a single dose of bovine casein hydrolysate (bCNH). bCNH22 - dairy cows were treated with two doses of bovine casein hydrolysate (bCNH). **p<0.01. Control 1.91 (SE) bCNH11: 1.94, bCNH22: 1.97; [Figure 8]This graph illustrates the monthly lactose yield (kg) in dairy cows treated with bovine casein hydrolysate (bCNH) during the dry period over a 305-day subsequent lactation period. Control cows were treated with antibiotics. bCNH11 cows were treated with a single dose of bovine casein hydrolysate (bCNH). bCNH22 cows were treated with two doses of bovine casein hydrolysate (bCNH). **p<0.01; [Figure 9] This is a monthly diagram of ECM. The mean monthly ECM before participation in the study was used as a covariate. The energy-adjusted milk yield for the control group over the entire period after resuming milking was 35.88 (SE=4.06), and the energy-adjusted milk yield for the bCNH group was 39.86 (SE=2.76), with a P=0.43. [Figure 10] This graph shows the monthly protein values ​​(kg). The average monthly protein yield before participation in the study was used as a covariate. The protein yield of the control group over the entire 3-month period after the start of milking was 1.32 kg (SE=0.098), and the protein yield of the bCNH group was 1.38 kg (SE=0.065), with a P=0.08. [Figure 11] This graph shows the monthly milk fat content (kg). The mean monthly milk fat content before participation in the study was used as a covariate. The milk fat yield of the control group was 1.36 kg (SE=0.14), and the milk fat yield of the bCNH group was 1.49 kg (SE=0.09) (*P=0.46). [Figure 12] This graph shows monthly lactose levels (kg). The average monthly lactose yield before participation in the study was used as a covariate. The least squares mean of lactose yield in the control group was 1.82 (SE=0.18), and in the bCNH group it was 1.90 (SE=0.12). [Figure 13]This graph shows the monthly composite bulk milk somatic cell count (BMSCC) during the pre- and post-re-milking periods. The bulk milk somatic cell count does not follow a normal distribution (Prob>chiSq=0.026). Therefore, the BMSCC values ​​were converted to log10. The difference at each lactation age (MIM) revealed that the log10 BMSCC of the control cows was significantly higher than that of the experimental group in the first month after the start of milking (*P=0.005). [Figure 14] This graph shows the monthly milk yield values ​​(kg) for each month. The average monthly milk yield before participation in the study was used as a covariate. The average monthly milk yield for the control group over the entire period after resuming milking was 39.27 kg (SE=3.56), and the average monthly milk yield for the bCNH group was 39.92 kg (SE=2.37), with a P=0.88.

[0025] For the sake of simplicity and clarity in the illustrations, it should be understood that the elements shown in the drawings are not necessarily drawn to scale. For example, the dimensions of some elements may be exaggerated relative to others for clarity. Furthermore, where deemed appropriate, reference numbers may be repeated between drawings to indicate corresponding or similar elements. [Modes for carrying out the invention]

[0026] In the following detailed description, numerous specific details are given for a thorough understanding of the invention. However, it will be understood by those skilled in the art that the invention can be carried out without these specific details. In other examples, well-known methods, procedures, and components are not described in detail so as not to obscure the invention.

[0027] Methods to improve milk quality In some embodiments, disclosed herein are methods for improving milk quality in lactating mammals, comprising administering a composition comprising at least one casein-derived peptide.

[0028] In some embodiments, improved milk quality includes increased milk protein, increased milk fat, increased lactose, increased energy-corrected milk (ECM), or a combination thereof. In one embodiment, improved milk quality includes increased milk protein. In another embodiment, improved milk quality includes increased milk fat. In another embodiment, improved milk quality includes increased lactose. In another embodiment, improved milk quality includes increased energy-corrected milk (ECM). In another embodiment, improved milk quality includes increased milk protein and increased milk fat. In another embodiment, improved milk quality includes increased milk protein and increased lactose. In another embodiment, improved milk quality includes increased milk protein and increased ECM. In another embodiment, improved milk quality includes increased milk fat and increased lactose. In another embodiment, improved milk quality includes increased milk fat and increased ECM. In another embodiment, improved milk quality includes increased lactose and increased ECM. In another embodiment, the improved milk quality includes increased milk protein, increased milk fat, increased lactose, and increased energy-corrected milk (ECM).

[0029] Those skilled in the art will understand that energy-corrected milk (ECM) determines the amount of energy in milk based on the milk, fat, and protein it contains. The determination of ECM can be carried out, for example, by following the following equation: ECM (kg) = Milk volume (kg) × 0.1 + Fat (%) × Milk volume (kg) × 10 + Protein (%) × Milk volume (kg) × 15.8.

[0030] In one embodiment, disclosed herein is a method for increasing milk protein in lactating mammals, comprising administering a composition comprising at least one casein-derived peptide. In one embodiment, the milk protein is extracted from milk.

[0031] In one embodiment, disclosed herein is a method for increasing milk fat in lactating mammals, comprising administering a composition comprising at least one casein-derived peptide. In one embodiment, milk fat is extracted from milk.

[0032] In one embodiment, disclosed herein is a method for increasing lactose in lactating mammals, comprising administering a composition comprising at least one casein-derived peptide. In one embodiment, lactose is extracted from milk.

[0033] In one embodiment, disclosed herein is a method for increasing extracorporeal membrane clotting (ECM) in lactating mammals, comprising administering a composition comprising at least one casein-derived peptide.

[0034] In some embodiments, the percentage of milk protein in the milk is in the range of 2.5% to 6.5%. In one embodiment, the percentage of milk protein in the milk is 2.5%. In another embodiment, the percentage of milk protein in the milk is 3.0%. In another embodiment, the percentage of milk protein in the milk is 3.5%. In another embodiment, the percentage of milk protein in the milk is 4.0%. In another embodiment, the percentage of milk protein in the milk is 4.5%. In another embodiment, the percentage of milk protein in the milk is 5.0%. In another embodiment, the percentage of milk protein in the milk is 5.5%. In another embodiment, the percentage of milk protein in the milk is 6.0%. In another embodiment, the percentage of milk protein in the milk is 6.5%.

[0035] In some embodiments, the proportion of milk protein in the milk is increased by 0.5% to 30.0% compared to the milk protein in a control lactating mammal. In some embodiments, the milk is increased by 1.0% to 20.0% compared to the milk protein in a control lactating mammal. In some embodiments, the milk is increased by 3.0% to 10.0% compared to the milk protein in a control lactating mammal. In some embodiments, the milk is increased by 5.0% to 8.0% compared to the milk protein in a control lactating mammal.

[0036] In one embodiment, the proportion of milk protein in the milk is increased by 0.5% compared to the milk protein in a control lactating mammal. In another embodiment, the proportion of milk protein in the milk is increased by 1.0% compared to the milk protein in a control lactating mammal. In another embodiment, the proportion of milk protein in the milk is increased by 5.0% compared to the milk protein in a control lactating mammal. In another embodiment, the proportion of milk protein in the milk is increased by 10.0% compared to the milk protein in a control lactating mammal. In another embodiment, the proportion of milk protein in the milk is increased by 15.0% compared to the milk protein in a control lactating mammal. In another embodiment, the proportion of milk protein in the milk is increased by 20.0% compared to the milk protein in a control lactating mammal. In another embodiment, the proportion of milk protein in the milk is increased by 25.0% compared to the milk protein in a control lactating mammal. In another embodiment, the proportion of milk protein in the milk is increased by 30.0% compared to the milk protein in a control lactating mammal.

[0037] In some embodiments, the percentage of milk fat in the milk is in the range of 2.5% to 6.5%. In one embodiment, the percentage of milk fat in the milk is 2.5%. In another embodiment, the percentage of milk fat in the milk is 3.0%. In another embodiment, the percentage of milk fat in the milk is 3.3%. In another embodiment, the percentage of milk fat in the milk is 3.4%. In another embodiment, the percentage of milk fat in the milk is 3.5%. In another embodiment, the percentage of milk fat in the milk is 3.6%. In another embodiment, the percentage of milk fat in the milk is 3.7%. In another embodiment, the percentage of milk fat in the milk is 3.8%. In another embodiment, the percentage of milk fat in the milk is 3.9%. In another embodiment, the percentage of milk fat in the milk is 4.0%. In another embodiment, the percentage of milk fat in the milk is 4.5%. In another embodiment, the percentage of milk fat in the milk is 5.0%. In another embodiment, the percentage of milk fat in the milk is 5.5%. In another embodiment, the percentage of milk fat in the milk is 6.0%. In another embodiment, the percentage of milk fat in the milk is 6.5%.

[0038] In some embodiments, the percentage of milk fat in the milk is increased by 0.5% to 30% compared to the milk fat in a control lactating mammal. In some embodiments, the percentage of milk fat in the milk is increased by 1% to 15% compared to the milk fat in a control lactating mammal. In some embodiments, the percentage of milk fat in the milk is increased by 2% to 6% compared to the milk fat in a control lactating mammal. In some embodiments, the percentage of milk fat in the milk is increased by 3% to 5% compared to the milk fat in a control lactating mammal.

[0039] In one embodiment, the percentage of milk fat in the milk is increased by 0.5% compared to the milk fat in a control lactating mammal. In another embodiment, the percentage of milk fat in the milk is increased by 1.0% compared to the milk fat in a control lactating mammal. In another embodiment, the percentage of milk fat in the milk is increased by 5.0% compared to the milk fat in a control lactating mammal. In another embodiment, the percentage of milk fat in the milk is increased by 10.0% compared to the milk fat in a control lactating mammal. In another embodiment, the percentage of milk fat in the milk is increased by 15.0% compared to the milk fat in a control lactating mammal. In another embodiment, the percentage of milk fat in the milk is increased by 20.0% compared to the milk fat in a control lactating mammal. In another embodiment, the percentage of milk fat in the milk is increased by 25.0% compared to the milk fat in a control lactating mammal. In another embodiment, the percentage of milk fat in the milk is increased by 30.0% compared to the milk fat in a control lactating mammal.

[0040] In some embodiments, the lactose content in milk is in the range of 3.5% to 7.8%. In one embodiment, the lactose content in milk is 3.5%. In another embodiment, the lactose content in milk is 4.0%. In another embodiment, the lactose content in milk is 4.5%. In another embodiment, the lactose content in milk is 5.0%. In another embodiment, the lactose content in milk is 5.5%. In another embodiment, the lactose content in milk is 6.0%. In another embodiment, the lactose content in milk is 6.5%. In another embodiment, the lactose content in milk is 7.0%. In another embodiment, the lactose content in milk is 7.5%. In another embodiment, the lactose content in milk is 7.8%.

[0041] In some embodiments, the proportion of lactose in milk is increased by 0.5% to 30% compared to the lactose in control lactating mammals. In some embodiments, the proportion of lactose in milk is increased by 1% to 15% compared to the lactose in control lactating mammals. In some embodiments, the proportion of lactose in milk is increased by 2% to 6% compared to the lactose in control lactating mammals. In some embodiments, the proportion of lactose in milk is increased by 3% to 5% compared to the lactose in control lactating mammals.

[0042] In one embodiment, the proportion of lactose in milk is increased by 0.5% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in milk is increased by 1% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in milk is increased by 1.5% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in milk is increased by 2.0% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in milk is increased by 2.5% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in milk is increased by 3.0% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in milk is increased by 3.5% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in milk is increased by 4.0% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in milk is increased by 4.5% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in milk is increased by 5.0% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in milk is increased by 5.5% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in milk is increased by 6.0% compared to the lactose in a control lactating mammal.

[0043] In another embodiment, the proportion of lactose in the milk is increased by 10% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in the milk is increased by 15% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in the milk is increased by 20% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in the milk is increased by 25% compared to the lactose in a control lactating mammal. In another embodiment, the proportion of lactose in the milk is increased by 30% compared to the lactose in a control lactating mammal.

[0044] In some embodiments, the ECM is increased by 0.5% to 20% compared to the ECM in a control lactating mammal. In some embodiments, the ECM is increased by 2.0% to 15% compared to the ECM in a control lactating mammal. In some embodiments, the ECM is increased by 5.0% to 10% compared to the ECM in a control lactating mammal. In some embodiments, the ECM is increased by 7.5% to 9% compared to the ECM in a control lactating mammal.

[0045] In some embodiments, milk quality further includes a decrease in somatic cell count (SCC). In one embodiment, the decrease in SCC occurs after the resumption of milking. In one embodiment, the decrease in SCC is observed in treated subjects compared to untreated subjects.

[0046] In some embodiments, milk quality is improved at any stage of the lactation cycle. Those skilled in the art will understand that “lactation cycle” means the period between one calving and the next. The cycle is divided into stages: early lactation, mid-lactation, late lactation, and dry period.

[0047] In one embodiment, milk quality is improved during early lactation, mid-lactation, late lactation, or any combination thereof. In another embodiment, milk quality is improved during early lactation. In another embodiment, milk quality is improved during mid-lactation. In another embodiment, milk quality is improved during late lactation. In another embodiment, milk quality is improved during early and mid-lactation. In another embodiment, milk quality is improved during early and late lactation. In another embodiment, milk quality is improved during mid and late lactation. In another embodiment, milk quality is improved during early, mid, and late lactation.

[0048] In some embodiments, milk quality is improved during the same lactation period following administration of the composition containing at least one casein-derived peptide. In some embodiments, milk quality is improved during the next lactation period following administration of the composition containing at least one casein-derived peptide.

[0049] In some embodiments, administration occurs at the start of the dry period. In some embodiments, administration occurs during lactation.

[0050] Those skilled in the art will understand that the "dry period" refers to the pre-calving period during which dairy cows are not milked, which currently lasts about 6-9 weeks. The dry period serves multiple functions. Its primary functions are to give the cow a rest period before the birth of the next calf and to maximize milk yield in the next lactation. During the dry period, mammary gland cells regenerate at a faster rate than when the cow would be milked until calving. At the start of the dry period, many dairy farmers administer antibiotics to treat the cows, especially in cases of persistent subclinical mastitis.

[0051] In some embodiments, milk quality is improved regardless of the length of the dry period. In one embodiment, the dry period includes 28 to 120 days. In another embodiment, the dry period includes 28 to 100 days. In yet another embodiment, the dry period includes 28 to 80 days. In yet another embodiment, the dry period includes 28 to 60 days. In yet another embodiment, the dry period includes 28 to 40 days. In yet another embodiment, the dry period includes 40 to 49 days. In yet another embodiment, the dry period includes 50 to 59 days.

[0052] In some embodiments, the milk is essentially free of residue.

[0053] In some embodiments, milk may be used as raw milk for dairy production, lactation, bread making, confectionery making, feeding, or a combination thereof. In one embodiment, milk may be used as raw milk. In another embodiment, milk may be used for dairy production. In another embodiment, milk may be used for lactation. In another embodiment, milk may be used for bread making. In another embodiment, milk may be used for confectionery making. In another embodiment, milk may be used for feeding.

[0054] In another embodiment, milk may be used for protein extraction. In another embodiment, milk may be used for milk fat extraction. In another embodiment, milk may be used for lactose extraction.

[0055] In one embodiment, the dairy product includes milk, whey, yogurt, cheese, cream, butter, a high-protein dairy beverage, or a combination thereof. In another embodiment, the dairy product includes milk. In another embodiment, the dairy product includes whey. In another embodiment, the dairy product includes yogurt. In another embodiment, the dairy product includes cheese. In another embodiment, the dairy product includes cream. In another embodiment, the dairy product includes butter. In another embodiment, the dairy product includes a high-protein dairy beverage.

[0056] In some embodiments, dairy products contain a high proportion of protein.

[0057] In one embodiment, the product obtained from high-quality milk is for use in the treatment of disease. In one embodiment, the product obtained from high-quality milk is for use in building and repairing muscle, building and repairing skin, building and repairing other body tissues, treating infections, balancing bodily fluids, transporting oxygen throughout the body, supporting cancer treatment, or any combination thereof.

[0058] In some embodiments, products obtained from high-quality milk may be used for non-food applications. In one embodiment, non-food applications include use in several technical applications such as protective coatings and foams, in the manufacture of plastics and other solid materials, in the production of fibers, adhesives, ethanol or methane, in research fields as barriers to nonpolar substances such as oxygen, carbon dioxide and aromas, in paper coatings, adhesives or injection-molded disposable products, as emulsifiers, as surfactants, in drug delivery, or any combination thereof.

[0059] In some embodiments, disclosed herein are compositions comprising at least one casein-derived peptide for use in improving milk quality in lactating mammals.

[0060] Casein peptide In some embodiments, the described method involves administering a composition comprising at least one casein-derived peptide.

[0061] Casein is a protein found in the milk of non-human mammals and also in the milk of human mammals, and is known to contain subgroups αS1, αS2, β, and κ. Casein is defined according to the respective amino acid sequences of subgroups αS1, αS2, β, and κ. In the context of this disclosure, when casein is referred to, it is understood to also include acid casein, salts of casein, phosphorus-containing casein, and rennet casein.

[0062] As used herein, the term “protein” refers to amino acid residues linked by peptide bonds. Protein sequences are generally reported from the N-terminus, which contains a free amino group, to the C-terminus, which contains a free carboxyl group. As used herein, amino acids refer to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function similarly to naturally occurring amino acids. Amino acids may be referred herein by their commonly known three-letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Committee.

[0063] The casein-derived peptide may be a single peptide or a mixture of different peptides that can be independently selected from naturally occurring peptides, semi-synthetic peptides, synthetic peptides, or recombinant peptides. It should be further noted that the peptides according to this disclosure may be produced synthetically, by recombinant DNA technology, or by any other technology. Methods for producing peptides are well known in the art.

[0064] In some embodiments, the casein-derived peptide may contain casein proteolytic products (also known in the art by the term "casein hydrolysate") that are produced when the casein protein is cleaved into peptide fragments by an enzyme or acid. Casein hydrolysate is understood as a hydrolyzed form of casein (protein). Casein hydrolysate includes, for example, active beta-, alpha-S1-, alpha-S2-, and kappa-casein-derived peptides known in the art. In some embodiments, the casein-derived peptide is or contains casein hydrolysate.

[0065] In some embodiments, the casein-derived peptides include natural peptides, synthetic peptides, semi-synthetic peptides, or any combination thereof. In another embodiment, the casein-derived peptides include natural peptides. In another embodiment, the casein-derived peptides include synthetic peptides. In another embodiment, the casein-derived peptides include semi-synthetic peptides. In another embodiment, the casein-derived peptides include a combination of natural and synthetic peptides. In another embodiment, the casein-derived peptides include a combination of natural and semi-synthetic peptides. In another embodiment, the casein-derived peptides include a combination of synthetic and semi-synthetic peptides. In another embodiment, the casein-derived peptides include a combination of natural peptides, synthetic peptides, and semi-synthetic peptides.

[0066] Peptides derived from natural casein are typically obtained following enzymatic hydrolysis, and the enzyme may be any mammalian peptidase, including, but not limited to, plasmin, pancreatin, trypsin, chymotrypsin, neutrase, alcalase, pepsin, carboxypeptidase, cathepsin, as well as, but not limited to, plant peptidases such as papain, bromelain, and enzymes derived from microorganisms. For example, naturally occurring casein peptides may be the result of enzymatic activity, such as plasmin, on casein subunits such as β-casein, αs1- and αs2-casein, or κ-casein. In some embodiments, the casein hydrolysate is obtained by cleavage of the casein protein with trypsin.

[0067] Synthetic peptides may be obtained by any method known in the industry for peptide synthesis, including chemical synthesis and recombinant DNA techniques. For example, peptides may be synthesized using standard solid-phase techniques.

[0068] In one embodiment, the synthetic peptide is a recombinant peptide.

[0069] Semi-synthetic casein-derived peptides may be obtained by chemical hydrolysis of casein, for example, by prolonged boiling in a strong acid (acid-HVP), or by prolonged boiling in a strong base, or by using a chemical agent such as cyanide bromide (CNBr). Casein-derived peptides may also be obtained by molecular engineering, for example, by using recombinant DNA in molecular techniques known in the art. In such embodiments, the casein-derived peptide is a recombinant peptide.

[0070] In one embodiment, recombinant peptides are produced by fermentation, tissue culture, or a combination thereof. In another embodiment, recombinant peptides are produced by fermentation. In yet another embodiment, recombinant peptides are produced by tissue culture. In yet another embodiment, recombinant peptides are produced by a combination of fermentation and tissue culture.

[0071] In one embodiment, the tissue culture includes mammary bovine tissue.

[0072] In some embodiments, the casein-derived peptide comprises one or more fragments of β-casein, αS1-casein, αS2-casein, κ-casein, or any combination thereof. In one embodiment, the casein-derived peptide comprises one or more fragments of β-casein. In another embodiment, the casein-derived peptide comprises one or more fragments of αS1-casein. In yet another embodiment, the casein-derived peptide comprises one or more fragments of αS2-casein. In yet another embodiment, the casein-derived peptide comprises one or more fragments of κ-casein. In yet another embodiment, the casein-derived peptide comprises one or more fragments of a combination of β-casein, αS1-casein, αS2-casein, and κ-casein.

[0073] In one embodiment, the casein-derived peptide further comprises amino acids having different lengths.

[0074] In one embodiment, the casein-derived peptide includes a casein hydrolysate.

[0075] In one embodiment, the casein-derived peptide includes a phosphopeptide.

[0076] As used herein, the term "phosphopeptide" refers to a phosphorylated peptide in the form of a conjugated peptide, where the non-peptide portion is a phosphate residue. The expression "phosphopeptide" or "phosphoserine" refers to a conjugated serine, where the non-peptide portion is a phosphate residue.

[0077] In some embodiments, the casein-derived peptide is a single peptide or a mixture of phosphopeptides, which contain a single phosphorus group or are phosphorus-fortified peptides. In some embodiments, the casein-derived peptide may be a phosphoserine, phosphotyrosine, phosphothreonine, and / or phosphohistidine-fortified casein-derived peptide (casein phosphopeptide, CPP) and a monovalent cation phosphocaseinate, such as sodium, potassium, calcium, or ammonium phosphocaseinate.

[0078] In some embodiments, the casein-derived peptide is a phosphate peptide.

[0079] Phosphatepeptides may be genetically engineered casein-derived peptides, as well as peptidomimetics of casein-derived peptides. For example, phosphorylation of an amino acid, such as at least one serine residue, may be carried out by any method known in the art. The term “casein-derived peptide” also includes peptide fragments or peptidomimetics products obtained from or corresponding to one or more parts of a casein protein. Peptidomimetics peptides may be, for example, peptoids or semipeptoids, which are peptide analogs and have modifications such as cyclization, N-terminal modification, C-terminal modification, peptide bond modification (including but not limited to CH2-NH, CH2-S, CH2-SO, OC-NH, CH2-O, CH2-CH2, SC-NH, CH-CH, or CF-CH), backbone modification, and residue modification.

[0080] As used herein, the term “casein-derived peptide” further encompasses any peptide of any derivative, analog, variant, or homolog. The term “derivative” is used to define an amino acid sequence (peptide) having any insertion, deletion, substitution, and modification to the amino acid sequence (peptide) that does not alter the activity of the original peptide. The term “derivative” also refers to homologs, variants, and analogs, as well as covalent modifications of polypeptides produced according to the present invention.

[0081] In some embodiments, modified, synthetic, semi-synthetic, or other types of analogs of naturally occurring casein-derived peptides are, in some embodiments, at least 75%, sometimes 85%, 90%, 95%, and even 99% identical (in sequence) to naturally occurring casein-derived peptides, provided that the two sequences are optimally aligned. Furthermore, any non-naturally occurring casein-derived peptides used in accordance with this disclosure may retain at least some of the biological activity of naturally occurring casein proteins.

[0082] This disclosure also includes homologs of casein-derived peptides. The term “homolog” is used to define an amino acid sequence (peptide) that maintains minimal homology to the amino acid sequences defined in this invention, for example, having at least about 65%, at least about 75%, at least about 85%, or at least about 95% overall sequence homology to any of the amino acid sequences of any particular sequence of the peptides structurally defined above.

[0083] In some embodiments, casein-derived peptides may also include chemical modifications of naturally occurring peptides, for example, in which one or more amino acids are deleted, substituted, or modified, for example, by side chain removal, side chain substitution, or introduction of a chemical group. Chemical modifications may include, but are not limited to, acetylation, acylation, amidation, ADP-ribosylation, glycosylation, GPI anchoring, covalent attachment of lipids or lipid derivatives, methylation, myristoylation, PEGylation, prenylation, phosphorylation, ubiquitination, or any similar process. Where referring to the substitution of an amino acid sequence by another, the substitution is likely to be a conserved substitution. For example, one or more amino acid residues in a casein sequence may be substituted by another amino acid having a similar polarity or charge. For example, nonpolar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine. Polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine. Positively charged (basic) amino acids include arginine, lysine, and histidine. Negatively charged (acidic) amino acids include aspartic acid and glutamic acid. Nevertheless, non-conservative substitutions may also occur, as long as they do not significantly alter the desired (casein-like) bioactivity of the casein-derived peptide analog from which they are produced.

[0084] The casein-derived peptides described herein are characterized by molecular weights of approximately 100 to 10,000 daltons on average (e.g., 2 to 100 amino acids), sometimes approximately 100 to 7,000 daltons on average, and sometimes 1,000 to 5,000 daltons on average.

[0085] The casein-derived peptides described herein are characterized by lengths of 2 to 200, 2 to 100 amino acids, sometimes 4 to 40 amino acids, sometimes 4 to 30 amino acids, sometimes 4 to 10 amino acids, and sometimes 10 to 50 amino acids.

[0086] In some embodiments, the casein-derived peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 26.

[0087] In one embodiment, the phosphopeptide comprises an amino acid sequence represented as Ser-Ser-Ser-Glu (SEQ ID NO: 1), where at least one Ser residue, at least two Ser residues, or three Ser residues are phosphorylated (phosphorylated serine is represented herein as Ser(p) or S(p)).

[0088] In another embodiment, the phosphopeptide comprises an amino acid sequence represented as Ser-Ser-Ser-Glu-Glu (SEQ ID NO: 2), where at least one Ser residue, at least two Ser residues, or three Ser residues are phosphorylated.

[0089] In another embodiment, the phosphopeptide comprises an amino acid sequence represented as Ser(p)-Ser(p)-Ser(p)-Glu-Glu (SEQ ID NO: 3).

[0090] In another embodiment, the phosphopeptide comprises an amino acid sequence represented as RELEELNVPGEIVES(p)LS(p)S(p)S(p)EESITR (SEQ ID NO: 4). In another embodiment, the phosphopeptide comprises an amino acid sequence represented as QMEAESIS(p)S(p)S(p)EEIVPDSVEQK (SEQ ID NO: 5). In another embodiment, the phosphopeptide comprises an amino acid sequence represented as KNTMEHVS(p)S(p)S(p)EESIISNETYK (SEQ ID NO: 6). In another embodiment, the phosphopeptide comprises an amino acid sequence represented as KVNELSKNIGS(p)ES(p)TEDQ (SEQ ID NO: 7). In another embodiment, the phosphopeptide comprises an amino acid sequence represented as PTLNREQLS(p)TS(p)EENSKKTVD (SEQ ID NO: 8). In another embodiment, the phosphopeptide comprises an amino acid sequence represented as ELEELNVPGEIVES(p)LS(p)S(p)S(p)EESITR (SEQ ID NO: 9). In another embodiment, the phosphopeptide comprises an amino acid sequence represented as RELEELNVPGEIES(p)LS(p)S(p)S(p)EESITR (SEQ ID NO: 10). In another embodiment, the phosphopeptide comprises an amino acid sequence represented as QMEAES(p)IS(p)S(p)S(p)EEIVPNS(p)VEQK (SEQ ID NO: 11). In another embodiment, the phosphopeptide comprises an amino acid sequence represented as KNTMEHVS(p)S(p)S(p)EESIIS(p)QETYK (SEQ ID NO: 12). In another embodiment, the phosphopeptide comprises an amino acid sequence represented as KVNELSKDIGS(p)ES(p)TEDQ (SEQ ID NO: 13). In another embodiment, the phosphopeptide comprises an amino acid sequence represented as ESIIS(p)QETYKQEKNMAINPSKENLCSTFCKEVVRNANEEETSIGS(p)S(p)S(p)EES(p)AEVATEEVKITVDDKHYQKALNEINQFYQKFPGYLQYLYQGPIVLNPWNQVLRNAVPITPTLNREQLS(p)TS(p)EENSKKTVN (Sequence ID 14). In another embodiment, the phosphopeptide comprises an amino acid sequence represented as ELEELNVPGEIES(p)LS(p)S(p)S(p)EESITR (Sequence ID 15).

[0091] In another embodiment, the phosphopeptide comprises an amino acid sequence represented as X1(n)-Ser(P)-Ser(P)-Ser(P)-X2(m)-Lys (SEQ ID NO: 16), where at least one of X1 and X2 is independently selected from positively charged amino acids, and each of n and m is independently selected from 0, 1, and 2.

[0092] In some embodiments, the positively charged amino acid is selected from the group consisting of lysine, arginine, and histidine. In some embodiments, the positively charged amino acid is lysine. In some other embodiments, the positively charged amino acid is arginine. In some other embodiments, the positively charged amino acid is histidine.

[0093] According to some embodiments, the formula of Sequence ID No. 16 further includes a blocking group (also referred to here as a protecting group) at the C-terminus. In some embodiments, the C-terminal carboxyl group of the peptide is protected with a protecting group. The protecting group is selected from, but is not limited to, amides (i.e., the C-terminal hydroxyl group is substituted with a primary amine (NH2), secondary amine, or tertiary amine) or esters (i.e., the C-terminal hydroxyl group is substituted with an ester). According to some embodiments, the blocking group is selected from the group consisting of amides and esters. According to some embodiments, the blocking group is an amide.

[0094] In another embodiment, the phosphopeptide includes an amino acid sequence represented as Lys-Lys-Ser(P)-Ser(P)-Ser(P) (SEQ ID NO: 17). In another embodiment, the phosphopeptide includes an amino acid sequence represented as Lys-Lys-Ser(P)-Ser(P)-Ser(P)-Lys (SEQ ID NO: 18). In another embodiment, the phosphopeptide includes an amino acid sequence represented as Lys-Lys-Ser(P)-Ser(P)-Ser(P)-Lys-Lys (SEQ ID NO: 19). In another embodiment, the phosphopeptide includes an amino acid sequence represented as Lys-Ser(P)-Ser(P)-Ser(P)-Lys-Lys (SEQ ID NO: 20). In another embodiment, the phosphopeptide includes an amino acid sequence represented as Lys-Ser(P)-Ser(P)-Ser(P)-Lys (SEQ ID NO: 21). In another embodiment, the phosphopeptide includes an amino acid sequence represented as Lys-Ser(P)-Ser(P)-Ser(P) (SEQ ID NO: 22). In another embodiment, the phosphopeptide includes an amino acid sequence represented as Ser(P)-Ser(P)-Ser(P)-Lys-Lys (SEQ ID NO: 23). In another embodiment, the phosphopeptide includes an amino acid sequence represented as Ser(P)-Ser(P)-Ser(P)-Lys (SEQ ID NO: 24). In another embodiment, the phosphopeptide includes an amino acid sequence represented as Lys-Lys-Ser(p)-Ser(p)-Ser(p)-NH2 (SEQ ID NO: 25). In another embodiment, the phosphopeptide includes an amino acid sequence represented as RELEELNVPGEIVES(p)LS(p)S(p)S(p)EESITRINK (SEQ ID NO: 26).

[0095] The casein-derived peptides according to the present invention may contain residues in both "L" and "D" forms. While all amino acid residues in the peptide sequences shown in SEQ ID NOs: 1-26 are in the "L" isomer form, the "D" isomer residues can be substituted for any L-amino acid residue, as long as the resulting peptide analog retains at least some of the biological activity of the corresponding "L" isomer. One reason for designing casein-derived peptides containing at least one D-amino acid is to increase the peptide's stability against protease degradation.

[0096] In one embodiment, the composition is free of antimicrobial agents and contains an acceptable carrier. In another embodiment, the composition further contains hormones, genetically modified organisms, or combinations thereof. Those skilled in the art will understand that a genetically modified organism (GMO) is an animal, plant, or microorganism whose DNA has been modified using genetic engineering techniques.

[0097] In one embodiment, milk-derived proteins are measured by UV in the range of 204-220 nm.

[0098] Dosage and administration In some embodiments, the method of the present disclosure includes administering 10 ng / ml to 500 mg / ml of milk-derived protein per dose. In one embodiment, the method of the present disclosure includes administering 1 mg / ml to 500 mg / ml of milk-derived protein per dose. In another embodiment, the method of the present disclosure includes administering 10 mg / ml to 450 mg / ml of milk-derived protein per dose. In another embodiment, the method of the present disclosure includes administering 50 mg / ml to 400 mg / ml of milk-derived protein per dose. In another embodiment, the method of the present disclosure includes administering 50 mg / ml to 70 mg / ml of milk-derived protein per dose. In another embodiment, the method of the present disclosure includes administering 100 mg / ml to 350 mg / ml of milk-derived protein per dose. In another embodiment, the method of the present disclosure includes administering 150 mg / ml to 300 mg / ml of milk-derived protein per dose. In another embodiment, the method of the present disclosure includes administering 200 mg / ml to 250 mg / ml of milk-derived protein per dose. In yet another embodiment, the method of the present disclosure includes administering 5 mg / ml to 30 mg / ml of milk-derived protein per dose.

[0099] In one embodiment, the milk-derived protein concentration in the composition is 0.1% to 30%. In another embodiment, the milk-derived protein concentration in the composition is 0.1%. In another embodiment, the milk-derived protein concentration in the composition is 1%. In another embodiment, the milk-derived protein concentration in the composition is 5%. In another embodiment, the milk-derived protein concentration in the composition is 10%. In another embodiment, the milk-derived protein concentration in the composition is 10%. In another embodiment, the milk-derived protein concentration in the composition is 15%. In another embodiment, the milk-derived protein concentration in the composition is 20%. In another embodiment, the milk-derived protein concentration in the composition is 25%. In another embodiment, the milk-derived protein concentration in the composition is 30%.

[0100] In one embodiment, administration includes oral, intraoral, topical, transdermal, subcutaneous, rectal, vaginal, parenteral, or any combination thereof. In another embodiment, administration includes oral administration. In another embodiment, administration includes oral administration. In another embodiment, administration includes topical administration. In another embodiment, administration includes transdermal administration. In another embodiment, administration includes subcutaneous administration. In another embodiment, administration includes rectal administration. In another embodiment, administration includes vaginal administration. In another embodiment, administration includes parenteral administration.

[0101] In another embodiment, administration includes intramammary injection into one or more nipples. In another embodiment, administration includes intramammary injection into one nipple. In another embodiment, administration includes intramammary injection into multiple nipples.

[0102] In some embodiments, the method of the present disclosure includes one to eight doses to a single nipple. In one embodiment, the method of the present disclosure includes one dose. In another embodiment, the method of the present disclosure includes two doses. In another embodiment, the method of the present disclosure includes three doses. In another embodiment, the method of the present disclosure includes four doses. In another embodiment, the method of the present disclosure includes five doses. In another embodiment, the method of the present disclosure includes six doses. In another embodiment, the method of the present disclosure includes seven doses. In another embodiment, the method of the present disclosure includes eight doses.

[0103] In some embodiments, the administration of this disclosure includes a subsequent administration. In one embodiment, the administration is performed immediately after the previous administration.

[0104] In some embodiments, the administration of the Disclosure includes intervals of about 1 hour to about 72 hours. In one embodiment, the administration includes intervals of about 1 hour. In one embodiment, the administration includes intervals of about 4 hours. In one embodiment, the administration includes intervals of about 5 hours. In one embodiment, the administration includes intervals of about 8 hours. In one embodiment, the administration includes intervals of about 10 hours. In one embodiment, the administration includes intervals of about 12 hours. In one embodiment, the administration includes intervals of about 15 hours. In one embodiment, the administration includes intervals of about 16 hours. In one embodiment, the administration includes intervals of about 20 hours. In one embodiment, the administration includes intervals of about 24 hours. In one embodiment, the administration includes intervals of about 25 hours. In one embodiment, the administration includes intervals of about 28 hours. In one embodiment, the administration includes intervals of about 30 hours. In one embodiment, the administration includes intervals of about 35 hours. In one embodiment, the administration includes intervals of about 36 hours. In one embodiment, the administration includes intervals of about 40 hours. In one embodiment, the administration includes intervals of about 45 hours. In one embodiment, the administration includes intervals of approximately 50 hours. In one embodiment, the administration includes intervals of approximately 55 hours. In one embodiment, the administration includes intervals of approximately 60 hours. In one embodiment, the administration includes intervals of approximately 65 hours. In one embodiment, the administration includes intervals of approximately 70 hours. In one embodiment, the administration includes intervals of approximately 72 hours.

[0105] In one embodiment, the administration includes intervals of approximately 1 hour to approximately 24 hours.

[0106] In one embodiment, administration takes place during the lactation period.

[0107] In one embodiment, administration includes continuous administration during the lactation period or multiple lactation periods. In another embodiment, administration includes continuous administration during the lactation period. In yet another embodiment, administration includes continuous administration during multiple lactation periods. In yet another embodiment, administration includes continuous administration during two lactation periods. In yet another embodiment, administration includes continuous administration during three lactation periods. In yet another embodiment, administration includes continuous administration during four lactation periods. In yet another embodiment, administration includes continuous administration during five lactation periods. In yet another embodiment, administration includes continuous administration during six lactation periods. In yet another embodiment, administration includes continuous administration during seven lactation periods. In yet another embodiment, administration includes continuous administration during eight lactation periods. In yet another embodiment, administration includes continuous administration during nine lactation periods. In yet another embodiment, administration includes continuous administration during ten lactation periods.

[0108] In one embodiment, administration includes continuous administration during the dry period or multiple dry periods. In another embodiment, administration includes continuous administration during the dry period. In yet another embodiment, administration includes continuous administration during multiple dry periods. In yet another embodiment, administration includes continuous administration during two dry periods. In yet another embodiment, administration includes continuous administration during three dry periods. In yet another embodiment, administration includes continuous administration during four dry periods. In yet another embodiment, administration includes continuous administration during five dry periods. In yet another embodiment, administration includes continuous administration during six dry periods. In yet another embodiment, administration includes continuous administration during seven dry periods. In yet another embodiment, administration includes continuous administration during eight dry periods. In yet another embodiment, administration includes continuous administration during nine dry periods. In yet another embodiment, administration includes continuous administration during ten dry periods.

[0109] Examples Example 1 - Casein hydrolysate administered intramammaryally as a treatment for dry cows. The objective of this randomized, blinded, controlled study was to evaluate the association between intramammary administration of bovine casein hydrolysate (bCNH) and subsequent lactation milk production, as well as subsequent protein, milk fat, and lactose yields. Milk production was measured by milk yield, energy-corrected milk (ECM), and a combination of protein, milk fat, and lactose yields. Local tolerability of the infusion and the patient's overall condition were also evaluated during subsequent lactation.

[0110] A total of 306 registered Israeli Holstein Friesian dairy cows were included in the analysis. Eligibility criteria included good general health, absence of clinical mastitis, or absence of other intramammary infections. Fourteen cows were excluded from the overall eligible registered cows in this study due to management events (5), miscarriage (6), and mammary condition (3). Cow registration followed the farm's normal introduction into the pre-dry period.

[0111] There were two bCNH experimental groups and one positive control group. The experimental groups included: (i) 109 dairy cows treated with a single syringe (20 mL of 1200 mg bCNH) (bCNH-11), and (ii) 96 dairy cows treated consecutively with two 20 mL syringes (40 mL) (bCNH-22). The control group, consisting of 101 cows, was treated with the conventional dry cow treatment (DCT) Nefpenzal® DC.

[0112] Analysis of milk quality-corrected milk yield (ECM) Milk Quality Corrected Milk Yield (ECM) determines the amount of energy in milk based on milk composition, specifically fat and protein. Calculating ECM is used to measure feed efficiency (FE), which is a cow's ability to convert dry matter feed into milk yield (kg), by dividing ECM by the amount of dry matter intake. Feed efficiency is used for genetic assessment and the effects of temperature and humidity indices.

[0113] The results of monthly ECM production (kg) in milk over the subsequent 305 days of lactation are shown in Figures 1 and 2. Analysis of the differences between each bCNH treatment group and the control revealed that bCNH11 and bCNH22 were significantly different from the control (P<0.05). ECM yield for bCNH11 was 3.07 kg higher per day than the control over the 305 days of lactation, and ECM yield for bCNH22 was 2.96 kg higher than the control. For the entire 305 days of lactation, the difference between the control and these bCNH groups was 965 kg per cow for bCNH11 and 902 kg per cow for bCNH22. The mean ECM yield of the bCNH treatment was higher than that of the control at all time points over the 305 days of lactation.

[0114] Analysis of milk fat yield The results of a linear mixed model for monthly milk fat yield (kg) over a 305-day subsequent lactation period are shown in Figures 3 and 4.

[0115] Analysis of protein yield The results of a linear mixed model for monthly protein yield (kg) over a 305-day subsequent lactation period are shown in Figures 5 and 6.

[0116] Analysis of lactose yield The results for monthly lactose yield over the subsequent lactation period of 305 days are shown in Figures 7 and 8.

[0117] The above results regarding the enhancement of energy-corrected milk and the biological components of milk (primarily milk protein, milk fat, and lactose) during lactation after drying out of dairy cows with bCNH are innovative and surprising findings. Treatment with bCNH had no negative impact on milk yield compared to antibiotics. Furthermore, the importance of higher milk fat and protein yields, later demonstrated by higher ECM, strengthens the idea that bCNH is a very effective dry-period treatment.

[0118] Example 2 - Field study to evaluate the effect of bovine casein hydrolysate on milk quality during lactation. The objective of this field study was to evaluate the clinical benefits of intramammary infusion of bCNH into the mammary compartment in terms of milk yield and milk quality during lactation and after the resumption of milking (after a 5-day milking cessation period following bCNH administration).

[0119] Twenty-three pregnant Israeli Holstein Friesian dairy cows of any number of lactations were enrolled after clinical examination to confirm subclinical mastitis, determined by elevated SCC and following two consecutive positive bacterial tests for NAS during the screening period (test days -7 and -6). On treatment day (Day 0), following clinical examination and morning milking, eligible cows (infected compartments, 1 or 2 compartments per cow) were randomized to receive bCNH or not treated (negative control). After bCNH treatment, treated udder compartments had a 6 (±1) day milking cessation period, followed by resumption of milking. Other compartments were either controls (untreated) or not included in the study (uninfected) and continued to be milked according to the normal milking management process. Milk samples were collected from infected udder sections on the day of treatment (D0), the day of resumption (D6±1), 12 hours later, the following morning's milking day, and 14 and 21 days after treatment for bacterial testing, somatic cell count (SCC), lactose, sodium:potassium ratio, and milk conductivity. Following treatment, all dairy cows were managed according to normal management procedures. All dairy cows were monitored for clinical signs of mastitis or other diseases until day 21 of the trial. Daily milk yield data was collected from the NOA system (Israeli Dairy Herd Management Program developed by the Israeli Dairy Board) until day 35 after treatment. In addition, normal monthly data on milk yield and milk components were collected for up to 3 months prior to and 3 months after the trial entry (treatment).

[0120] Each dairy cow in the study received treatment administered only to the infected udder section(s); that is, each infected section of each cow received either 1200 mg of bCNH (T2), 2400 mg of bCNH (T3), or 4800 mg of bCNH (T4), or received no treatment (T1).

[0121] The treated udder section(s) of each participating dairy cow were not milked for 6 consecutive days (±1) before re-milking began. Untreated sections of the treated cows were milked as usual, and their milk was not discarded.

[0122] The effects of the treatment on milk yield, energy-corrected milk (ECM), milk fat (kg), protein (kg), lactose (kg), and complex SCC were examined.

[0123] Monthly milk yield, ECM, milk fat, protein, lactose, and composite SCC data were collected from the three months prior to treatment of the cows and the three months after resumption of milking. A linear mixed model was used to analyze the differences between the control and active treatment groups using JMP 16.2.0, LSmeans difference Student's t. The model included calving number (1, 2, 3+), lactation age (months) after resumption of milking (MIM), treatment (control, T2), and interaction between treatment and calving number by MIM. The mean of each independent variable for the period prior to enrollment in the study was calculated and used as a covariate. Energy-corrected milk (ECM) values ​​(kg) before and 3 months after the re-milking period. Energy-adjusted milk (kg) was calculated according to the following formula: ECM = 0.1 * milk + milk * milk fat % / 100 * 10 * milk * protein % / 100 * 15.8. The mean monthly ECM before participation in the study was used as a covariate. The energy-adjusted milk yield for the entire period after resuming milking in the control group was 35.88 (SE=4.06), and the energy-adjusted milk yield for the bCNH group was 39.86 (SE=2.76), with P=0.43. A significant difference was observed in the second month, as can be seen in Figure 9.

[0124] Complex protein levels (kg) in milk before the re-milking period and for 35 days after the re-milking period. The average monthly protein yield before participation in the study was used as a covariate. The protein yield for the control group over the entire 3-month period after the start of milking was 1.32 kg (SE=0.098), while the protein yield for the bCNH group was 1.38 kg (SE=0.065), with a P=0.08. As can be seen in Figure 10, protein yield was higher in the second and third months.

[0125] Combined milk fat content (kg) in milk before the re-milking period and for 35 days after the re-milking period. The mean monthly milk fat levels prior to participation in the study were used as a covariate. The milk fat yield in the control group was 1.36 kg (SE=0.14), and in the bCNH group it was 1.49 kg (SE=0.09) (*P=0.46). As can be seen in Figure 11, milk fat yield was higher during the measurement periods of the second and third months.

[0126] Complex lactose levels (kg) in milk before the re-milking period and for 35 days after the re-milking period. The mean monthly lactose yield before participation in the study was used as a covariate. The least squares mean lactose yield in the control group was 1.82 (SE=0.18), and in the bCNH group it was 1.90 (SE=0.12). As can be seen in Figure 12, lactose yield was higher in the second and third months after treatment.

[0127] Compound bulk milk somatic cell count (BMSCC) during the pre- and post-re-milking periods. Bulk milk somatic cell counts do not follow a normal distribution (Prob>chiSq=0.026). Therefore, the BMSCC value is log 10 Converted to: Log after resuming milking, independent of pre-treatment level. 10 To understand the effect of the treatment on BMSCC, pre-treatment log 10 BMSCC was used as a covariate. The log of the control group over the entire period after re-milking was used. 10BMSCC is 5.58 (SE=0.18), and the log of the bCNH group 10 The BMSCC was 5.14 (SE=0.13) (P=0.051). The difference at each lactation age (MIM) is the log of the control cow. 10 BMSCC was the log of the experimental group in the first month after the start of milking. 10 The results were significantly higher than those of BMSCC (P=0.005), as revealed in Figure 13.

[0128] Monthly milk yield (kg) before re-milking and for 35 days after re-milking. The average energy-adjusted milk yield for the control group over the entire period after resuming milking was 39.27 kg per day (SE=3.56), while the average energy-adjusted milk yield for the bCNH group was 39.92 kg per day (SE=2.37, *P=0.88). As can be seen in Figure 14, a higher average milk yield was observed in the bCNH-treated group. Therefore, it is considered that the experimental (bCNH) group showed a tendency toward higher milk yield over time.

[0129] While certain features of the present invention are illustrated and described herein, many modifications, substitutions, alterations, and equivalents will now arise for those skilled in the art. It should therefore be understood that the appended claims are intended to encompass all such modifications and alterations that fall within the true spirit of the present invention.

Claims

1. A method for improving milk quality in lactating mammals, comprising administering a composition containing at least one casein-derived peptide.

2. The method according to claim 1, wherein the improved milk quality comprises increased milk protein, increased milk fat, increased lactose, increased energy-corrected milk (ECM), or any combination thereof.

3. The method according to claim 2, wherein the improved milk quality includes increased milk protein.

4. The method according to claim 2, wherein the improved milk quality includes increased milk fat.

5. The method according to claim 2, wherein the improved milk quality includes increased lactose.

6. The method according to claim 2, wherein the improved milk quality includes increased energy-corrected milk (ECM).

7. The method according to claims 2 and 3, wherein the proportion of milk protein in the milk is in the range of 2.5% to 6.5%.

8. The method according to claims 2 and 3, wherein the proportion of milk protein in the milk is increased by 0.5% to 30% compared to the milk protein in a control lactating mammal.

9. The method according to claim 8, wherein the proportion of milk protein in the milk is increased by 1% to 20% compared to the milk protein in a control lactating mammal.

10. The method according to claims 8 and 9, wherein the proportion of the milk protein in the milk is increased by 3% to 10% compared to the milk protein in a control lactating mammal.

11. The method according to claims 8 to 10, wherein the proportion of the milk protein in the milk is increased by 5% to 8% compared to the milk protein in a control lactating mammal.

12. The method according to claims 2 and 4, wherein the proportion of milk fat in the milk is in the range of 2.5% to 6.5%.

13. The method according to claims 2 and 4, wherein the proportion of milk fat in the milk is increased by 0.5% to 30% compared to the milk fat in a control lactating mammal.

14. The method according to claim 13, wherein the proportion of milk fat in the milk is increased by 1% to 15% compared to the milk fat in a control lactating mammal.

15. The method according to claims 13 and 14, wherein the proportion of milk fat in the milk is increased by 2% to 6% compared to the milk fat in a control lactating mammal.

16. The method according to claims 13 to 15, wherein the proportion of milk fat in the milk is increased by 3% to 5% compared to the milk fat in a control lactating mammal.

17. The method according to claims 2 and 5, wherein the proportion of lactose in the milk is in the range of 3.5 to 7.8%.

18. The method according to claims 2 and 5, wherein the proportion of lactose in the milk is increased by 0.5% to 30% compared to lactose in a control lactating mammal.

19. The method according to claim 18, wherein the proportion of lactose in the milk is increased by 1% to 15% compared to lactose in a control lactating mammal.

20. The method according to claim 18, wherein the proportion of lactose in the milk is increased by 2% to 6% compared to lactose in a control lactating mammal.

21. The method according to claims 18 to 20, wherein the proportion of lactose in the milk is increased by 3% to 5% compared to the lactose in a control lactating mammal.

22. The method according to claims 2 and 6, wherein the ECM is increased by 0.5% to 20% compared to the ECM in a control lactating mammal.

23. The method according to claim 22, wherein the ECM is increased by 2% to 15% compared to the ECM in a control lactating mammal.

24. The method according to claims 22 and 23, wherein the ECM is increased by 5% to 10% compared to the ECM in a control lactating mammal.

25. The method according to any of the prior claims, wherein the milk quality further comprises a decrease in somatic cell count (SCC).

26. The method according to any of the prior claims, wherein the milk quality is improved at any stage of the lactation cycle.

27. The method according to claim 26, wherein the milk quality is improved during early lactation, mid-lactation, late lactation, or any combination thereof.

28. The method according to claims 1 to 27, wherein the milk quality is improved during the same lactation period of administration of the composition comprising at least one casein-derived peptide.

29. The method according to claims 1 to 27, wherein the milk quality is improved during the lactation period following administration of the composition comprising at least one casein-derived peptide.

30. The method according to claim 29, wherein the administration is performed at the start of the dry period.

31. The method according to any of the prior claims, wherein the milk quality is improved independently of the length of the dry period.

32. The method according to claim 31, wherein the dry period includes 28 to 120 days.

33. The method according to claim 31, wherein the dry period includes 28 to 60 days.

34. The method according to claim 31, wherein the dry period includes 40 to 49 days.

35. The method according to claim 31, wherein the dry period includes 50 to 59 days.

36. The method according to any of the prior claims, wherein the casein-derived peptide includes a natural peptide, a synthetic peptide, a semi-synthetic peptide, or any combination thereof.

37. The method according to claim 36, wherein the synthetic peptide is a recombinant peptide.

38. The method according to claim 37, wherein the recombinant peptide is produced by fermentation, tissue culture, or a combination thereof.

39. The method according to claim 38, wherein the tissue culture comprises mammary bovine tissue.

40. The method according to any of the prior claims, wherein the casein-derived peptide comprises one or more fragments of β-casein, αS1-casein, αS2-casein, and κ-casein, and optionally further comprises amino acids having different lengths or any combination thereof.

41. The method according to any one of the prior claims, wherein the casein-derived peptide includes a casein hydrolysate.

42. The method according to any of the prior claims, wherein the casein-derived peptide includes a phosphopeptide.

43. The method of claim 42, wherein the phosphopeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to SEQ ID NOs:

26.

44. The method according to any of the prior claims, wherein the composition is free of an antimicrobial agent and includes an acceptable carrier.

45. The method according to any of the prior claims, comprising administering the casein-derived peptide in an amount of 10 ng / ml to 500 mg / ml.

46. The method according to any of the prior claims, wherein the administration comprises intramammary injection into one or more nipples.

47. The method according to any of the prior claims, wherein the administration comprises one to eight administrations per nipple.

48. The method according to any of the prior claims, wherein the administration includes an interval of about 1 hour to about 72 hours.

49. The method according to any of the prior claims, wherein the administration includes continuous administration during the lactation period or multiple lactation periods.

50. The method according to any of the prior claims, wherein the milk is substantially free of residues and can be used as raw milk for dairy product manufacturing, breastfeeding, bread making, confectionery making, feeding or any combination thereof.

51. The method according to claim 50, wherein the dairy product includes milk, whey, yogurt, cheese, cream, butter, a high-protein dairy beverage, or a combination thereof.

52. A composition comprising at least one casein-derived peptide for use in improving milk quality in lactating mammals.

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