Compositions and methods for modulating breast disorders and conditions

Abstract

The present invention provides compositions and methods for treating infections in the breasts of a lactating mammal during lactation, and provides continuous milk production and milking of a lactating mammal during lactation wherein somatic cell count in milk, milk conductivity, or a combination thereof is increased.

Description

[0001] Sequence Listing Statement

[0002] This application contains a sequence listing that has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. The XML copy, created on November 14, 2023, is named P-609343-PC-SL.XML and is 66,293 bytes in size. Background of the Invention

[0003] According to its electrophoretic mobility, casein contains three fractions, α, β, and κ. Casein hydrolysate is a hydrolyzed form of casein that includes bioactive β-casein-derived peptides. It has been determined that casein hydrolysate plays a role in the immune response against microbial and viral infections.

[0004] Intramammary infection (IMI) plays a decisive role in human breastfeeding and the dairy industry, affecting animal health, making it difficult to maintain milk quality and production, and having a negative impact on welfare and comfort, thereby causing considerable economic losses.

[0005] IMI is a costly disease and requires early diagnosis to reduce the negative impact on milk quality and quantity and to maximize the chances of cure and prevent transmission.

[0006] Mastitis is a disease usually caused by IMI and is caused by pathogens, mainly bacteria, but also yeasts, fungi, or even algae. Mastitis can be clinical, with local (and in some cases systemic) clinical signs and milk abnormalities, or subclinical, with production losses and reduced milk quality.

[0007] Antimicrobial treatments have been used to maintain a balance between cow udder health and economics. Conversely, the emergence and spread of antimicrobial resistance (AMR) is an urgent matter of public interest, and thus, the widespread use of antimicrobials (AMU) in human and animal health and in livestock production is being criticized.

[0008] Hygiene and management measures during milk collection may reduce but not control the incidence of both clinical and subclinical mastitis. Currently, many dairies are equipped with online computerized data collection systems that are designed to detect relevant parameters of milk and dairy cows. Therefore, there is a recognized need for a composition and method to manage IMI in the early stages of the mammalian lactation period, which would be highly advantageous. Summary of the Invention

[0009] In some aspects, the present disclosure provides a method for treating intramammary infections in lactating mammals during lactation, the method comprising administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the period of cure of the lactating mammal occurs during the same lactation. In some related aspects, the period of cure of the lactating mammal comprises about 60 days or fewer days after treatment. In some related aspects, the period of cure of the lactating mammal comprises about 45 days or fewer days after treatment. In some related aspects, the period of cure of the lactating mammal comprises about 14 days or fewer days after treatment. In some related aspects, the period of cure of the lactating mammal comprises about 7 days or fewer days after treatment.

[0010] In some related aspects, the period of cure enables the infected teat to resume milk production during the same lactation. In some additional related aspects, the period of cure enables the uninfected teats to continue milk production.

[0011] In some additional aspects, the present disclosure provides a method for continuous milk production and milking in a lactating mammal during lactation, wherein the lactating mammal has a subclinical or clinical disease of intramammary infection, the method comprising administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or multiple mammary regions.

[0012] In some additional aspects, the subclinical disease comprises an increase in somatic cell count in milk, an increase in milk conductivity, a decrease in milk volume, or a combination thereof. In some related aspects, an increase in somatic cell count in milk, an increase in milk conductivity, a decrease in milk volume, or any combination thereof is a result of an uninfected or infected mammary region.

[0013] In some related aspects, the lactating mammal continues to produce and milk from an untreated single or multiple mammary regions. In some additional related aspects, the milk is substantially free of residues and can be used as raw milk for dairy production, for breastfeeding, or any combination thereof. In some additional related aspects, dairy products include milk, whey, yogurt, cheese, cream, butter, milk beverages with high protein, or a combination thereof.

[0014] In some related aspects, the milk-derived protein comprises casein or casein-derived peptides.

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

[0016] In some additional related aspects, the casein-derived peptides comprise one or more fragments of β-casein, αS1-casein, αS2-casein, and κ-casein. In some additional related aspects, the casein-derived peptides further comprise amino acids of different lengths.

[0017] In some additional related aspects, the casein-derived peptide comprises a casein hydrolyzate. In some additional related aspects, the casein-derived peptide comprises a phosphopeptide. In some additional related aspects, the phosphopeptide comprises an amino acid sequence selected from SEQ ID NO.1 - SEQ ID NO.26. Detailed Description

[0018] In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. However, those skilled in the art will understand that the present invention may be practiced without these specific details. In other instances, well-known methods, procedures, and components have not been described in detail so as not to obscure the present invention.

[0019] Treatment Methods

[0020] In some embodiments, a method for treating intramammary infections in lactating mammals during lactation is disclosed herein, the method comprising administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the healing period of the lactating mammal occurs during the same lactation period.

[0021] In some embodiments, a composition comprising at least one milk-derived protein is disclosed herein for treating intramammary infections in lactating mammals during lactation, wherein the healing period of the lactating mammal occurs during the same lactation period.

[0022] One of ordinary skill in the art understands the term "cure" to mean treating, alleviating symptoms, relieving, altering, stopping, improving a condition (e.g., a disease), the symptoms of a condition, or affecting a condition (or disease), or preventing or delaying the onset of symptoms, complications, biochemical parameters of a disease, or otherwise stopping or inhibiting the further development of a disease or its symptoms.

[0023] In one embodiment, the healing period of the lactating mammal comprises 60 days or fewer days after treatment. In another embodiment, the healing period of the lactating mammal comprises 60 days after treatment. In one embodiment, the healing period of the lactating mammal comprises 55 days or fewer days after treatment. In another embodiment, the healing period of the lactating mammal comprises 55 days after treatment. In one embodiment, the healing period of the lactating mammal comprises 50 days or fewer days after treatment. In another embodiment, the healing period of the lactating mammal comprises 50 days after treatment. In another embodiment, the healing period of the lactating mammal comprises 45 days or fewer days after treatment. In one embodiment, the healing period comprises 45 days or fewer. In another embodiment, the healing period comprises 45 days. In another embodiment, the healing period comprises 40 days or fewer. In another embodiment, the healing period comprises 40 days. In another embodiment, the healing period comprises 30 days or fewer. In another embodiment, the healing period comprises 30 days. In another embodiment, the healing period comprises 21 days. In another embodiment, the healing period comprises 20 days. In another embodiment, the healing period comprises 18 days. In another embodiment, the healing period comprises 16 days. In another embodiment, the healing period comprises 14 days. In another embodiment, the healing period comprises 12 days. In another embodiment, the healing period comprises 10 days. In another embodiment, the healing period comprises 9 days. In another embodiment, the healing period comprises 8 days. In another embodiment, the healing period comprises 7 days. In another embodiment, the healing period comprises 6 days. In another embodiment, the healing period comprises 5 days. In another embodiment, the healing period comprises 4 days. In another embodiment, the healing period comprises 3 days. In another embodiment, the healing period comprises 2 days.

[0024] In some embodiments, the healing period may enable the infected teat to resume milk production during the same lactation period. In one embodiment, the resumption of milk production occurs within the same lactation period. In another embodiment, the resumption of milk production occurs during a subsequent lactation period. In another embodiment, the resumption of milk production begins from the treated single or multiple mammary regions.

[0025] In some embodiments, the resumption of milk production occurs within 60 days or less of treatment. In other embodiments, the resumption of milk production occurs within 45 days or less of treatment. In further embodiments, the resumption of milk production occurs within 30 days or less of treatment. In further embodiments, the resumption of milk production occurs within 14 days or less of treatment. In other embodiments, the resumption of milk production occurs within 7 days or less of treatment. In further embodiments, the resumption of milk production occurs within 5 days or less of treatment. In other embodiments, the resumption of milk production occurs within 3 days or less of treatment.

[0026] In one embodiment, the healing period enables uninfected teats to continue milk production.

[0027] In one embodiment, the milk production amount at the resumption of milk production is equal to or higher than the milk production amount before treatment. In another embodiment, the milk production amount at the resumption of milk production is equal to the milk production amount before treatment. In another embodiment, the milk production amount at the resumption of milk production is higher than the milk production amount before treatment. In another embodiment, the milk production amount at the resumption of milk production is higher than the milk production amount of lactating mammals treated with antibiotics.

[0028] In one embodiment, within 2 - 90 days after the resumption of milk production, the milk production amount at the resumption of milk production is equal to or higher than the milk production amount before treatment. In another embodiment, 2 days after the resumption of milk production, the milk production amount at the resumption of milk production is equal to or higher than the milk production amount before treatment. In another embodiment, 10 days after the resumption of milk production, the milk production amount at the resumption of milk production is equal to or higher than the milk production amount before treatment. In another embodiment, 30 days after the resumption of milk production, the milk production amount at the resumption of milk production is equal to or higher than the milk production amount before treatment. In another embodiment, 50 days after the resumption of milk production, the milk production amount at the resumption of milk production is equal to or higher than the milk production amount before treatment. In another embodiment, 70 days after the resumption of milk production, the milk production amount at the resumption of milk production is equal to or higher than the milk production amount before treatment. In another embodiment, 90 days after the resumption of milk production, the milk production amount at the resumption of milk production is equal to or higher than the milk production amount before treatment.

[0029] In one embodiment, the milk yield when milking restarts is 0.5% - 10% higher than the milk yield before treatment. In another embodiment, the milk yield when milking restarts is 2% higher than the milk yield before treatment. In another embodiment, the milk yield when milking restarts is 3% higher than the milk yield before treatment. In another embodiment, the milk yield when milking restarts is 4% higher than the milk yield before treatment. In another embodiment, the milk yield when milking restarts is 5% higher than the milk yield before treatment. In another embodiment, the milk yield when milking restarts is 6% higher than the milk yield before treatment. In another embodiment, the milk yield when milking restarts is 7% higher than the milk yield before treatment. In another embodiment, the milk yield when milking restarts is 8% higher than the milk yield before treatment. In another embodiment, the milk yield when milking restarts is 9% higher than the milk yield before treatment. In another embodiment, the milk yield when milking restarts is 10% higher than the milk yield before treatment.

[0030] In another embodiment, the milk yield when milking restarts is more than 10% higher than the milk yield before treatment.

[0031] In one embodiment, the milk production before treatment includes the milk production up to 60 days after treatment. In another embodiment, the milk production before treatment includes the milk production 60 days after treatment. In another embodiment, the milk production before treatment includes the milk production up to 30 days after treatment. In another embodiment, the milk production before treatment includes the milk production 30 days after treatment. In another embodiment, the milk production before treatment includes the milk production up to 25 days after treatment. In another embodiment, the milk production before treatment includes the milk production 25 days after treatment. In another embodiment, the milk production before treatment includes the milk production up to 20 days after treatment. In another embodiment, the milk production before treatment includes the milk production 20 days after treatment. In another embodiment, the milk production before treatment includes the milk production up to 15 days after treatment. In another embodiment, the milk production before treatment includes the milk production 15 days after treatment. In another embodiment, the milk production before treatment includes the milk production up to 14 days after treatment. In another embodiment, the milk production before treatment includes the milk production 14 days after treatment. In another embodiment, the milk production before treatment includes the milk production 12 days after treatment. In another embodiment, the milk production before treatment includes the milk production 10 days after treatment. In another embodiment, the milk production before treatment includes the milk production 8 days after treatment. In another embodiment, the milk production before treatment includes the milk production 7 days after treatment. In another embodiment, the milk production before treatment includes the milk production 6 days after treatment. In another embodiment, the milk production before treatment includes the milk production 5 days after treatment. In another embodiment, the milk production before treatment includes the milk production 4 days after treatment. In another embodiment, the milk production before treatment includes the milk production 3 days after treatment. In another embodiment, the milk production before treatment includes the milk production 2 days after treatment. In another embodiment, the milk production before treatment includes the milk production 1 day after treatment.

[0032] In some embodiments, disclosed herein is a method for continuous milking and milk production in a lactating mammal during the lactation period, wherein the lactating mammal is in a clinical disease state of intramammary infection, the method comprising administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the milking is from one or more untreated mammary regions.

[0033] In some embodiments, disclosed herein is a method for continuous milking and milk production in a lactating mammal during the lactation period, wherein the lactating mammal has a subclinical disease of intramammary infection, the method comprising administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the milking is from one or more untreated mammary regions.

[0034] Those skilled in the art will understand that the term "subclinical disease" refers to an intramammary infection, where the disease process ranges from the time of exposure susceptibility to the asymptomatic onset of the process, including an increase in somatic cell count in milk, an increase in milk electrical conductivity, and a decrease in milk volume or a combination thereof.

[0035] Those skilled in the art will understand that the term "intramammary infection (IMI)" refers to the presence of infectious organisms in the mammary gland. The terms IMI, clinical mastitis, and subclinical mastitis are used almost interchangeably. Intramammary infection (IMI) includes a group of costly diseases that affect animals (including humans) worldwide.

[0036] In some embodiments, the subclinical disease includes an increase in somatic cell count in milk, an increase in milk electrical conductivity, a decrease in milk volume, or any combination thereof. In one embodiment, the subclinical disease includes an increase in somatic cell count in milk. In another embodiment, the subclinical disease includes an increase in milk electrical conductivity. In another embodiment, the subclinical disease includes a decrease in milk volume. In another embodiment, the subclinical disease includes an increase in somatic cell count in milk, an increase in milk electrical conductivity, and a decrease in milk volume.

[0037] In one embodiment, an increase in somatic cell count in milk, an increase in milk electrical conductivity, a decrease in milk volume, or any combination thereof is the result of an uninfected or infected mammary region. In another embodiment, an increase in somatic cell count in milk is the result of an uninfected or infected mammary region. In another embodiment, an increase in milk electrical conductivity is the result of an uninfected or infected mammary region. In another embodiment, a decrease in milk volume is the result of an uninfected or infected mammary region. In another embodiment, an increase in somatic cell count in milk, an increase in milk electrical conductivity, and a decrease in milk volume are the result of an uninfected or infected mammary region.

[0038] In one embodiment, a method for continuously milking a lactating mammal during lactation is disclosed herein, wherein the somatic cell count in milk increases from a single region or multiple regions, the method comprising administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or multiple mammary regions.

[0039] In one embodiment, a method for continuously milking a lactating mammal during lactation is disclosed herein, wherein the milk electrical conductivity increases from a single region or multiple regions, the method comprising administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or multiple mammary regions.

[0040] In one embodiment, a method for continuous milk production and milking in a lactating mammal during lactation is disclosed herein, wherein the milk volume is reduced from a single area or multiple areas, and the method comprises administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or multiple mammary gland areas.

[0041] In some embodiments, a method for continuous milk production and milking in a lactating mammal during lactation is disclosed herein, wherein the somatic cell count, milk conductivity, or a combination thereof in the milk increases from a single area or multiple areas, and wherein the milk volume is reduced from a single area or multiple areas, and the method comprises administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or multiple mammary gland areas.

[0042] In one embodiment, the somatic cell count in the milk is about 100,000 cells / ml or more. In another embodiment, the increase in the somatic cell count in the milk is about 150,000 cells / ml or more. In another embodiment, the increase in the somatic cell count in the milk is about 180,000 cells / ml or more. In another embodiment, the increase in the somatic cell count in the milk is about 1 million cells / ml or more. In another embodiment, the increase in the somatic cell count in the milk is about 5 million cells / ml or more. In another embodiment, the increase in the somatic cell count in the milk is about 10 million cells / ml or more.

[0043] In some embodiments, a method for continuous milk production and milking in a lactating mammal during lactation is disclosed herein, wherein the somatic cell count and milk conductivity in the milk increase from a single area or multiple areas, and the method comprises administering to the lactating mammal a composition comprising at least one milk-derived protein.

[0044] In some embodiments, a method for continuous milk production and milking in a lactating mammal during lactation is disclosed herein, and the method comprises administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the comfort during milking is increased.

[0045] As used herein, the term "comfort" refers to the presence of prevention of distress and an increase in positive sensations, commonly referred to as comfort or pleasure, which results particularly from an increase in the lying-down period, an increase in rumination time, a reduction in metabolic requirements, a reduction in udder pressure and / or nipple leakage, a reduction in the incidence of mastitis and other diseases, and a reduction in the lameness effect due to high milk production.

[0046] In some embodiments, a method for continuous milk production and milking in a lactating mammal during the lactation period is disclosed herein, the method comprising administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the treatment period is at any stage of lactation, including drying one or more mammary regions.

[0047] Those skilled in the art understand the term "lactation period" in the dairy industry as the time period between one calving and the next calving.

[0048] In one embodiment, at 25 °C, the increase in milk conductivity is about 3.0 millisiemens (mS) or higher from single or multiple measurements taken at intervals of several hours to several months. In another embodiment, at 25 °C, the increase in milk conductivity is about 4 millisiemens (mS) or higher. In another embodiment, at 25 °C, the increase in milk conductivity is about 5 millisiemens (mS) or higher. In another embodiment, at 25 °C, the increase in milk conductivity is about 6 millisiemens (mS) or higher. In another embodiment, at 25 °C, the increase in milk conductivity is about 7 millisiemens (mS) or higher. In another embodiment, at 25 °C, the increase in milk conductivity is about 8 millisiemens (mS) or higher. In another embodiment, at 25 °C, the increase in milk conductivity is about 9 millisiemens (mS) or higher. In another embodiment, at 25 °C, the increase in milk conductivity is about 10 millisiemens (mS) or higher.

[0049] In one embodiment, a method for continuous milk production and milking in a lactating mammal during the lactation period is disclosed herein, wherein bacteriology is positive in a single region or multiple regions, the method comprising administering to the lactating mammal a composition comprising at least one milk-derived protein.

[0050] In one embodiment, the bacteriology is positive in single or multiple measurements. In one embodiment, the bacteriology is positive in a single region or multiple regions.

[0051] In one embodiment, positive bacteriology in a single area or multiple areas is associated with a decrease in milk. In one embodiment, milk production is reduced by at least 2% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In one embodiment, milk production is reduced by at least 10% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In one embodiment, milk production is reduced by 10 - 50% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In another embodiment, milk production is reduced by 10% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In another embodiment, milk production is reduced by 20% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In another embodiment, milk production is reduced by 23% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In another embodiment, milk production is reduced by 25% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In another embodiment, milk production is reduced by 30% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In another embodiment, milk production is reduced by 35% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In another embodiment, milk production is reduced by 40% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In another embodiment, milk production is reduced by 45% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In another embodiment, milk production is reduced by 50% compared to milk production resulting from negative bacteriology in a single area or multiple areas. In another embodiment, milk production is reduced by more than 50% compared to milk production resulting from negative bacteriology in a single area or multiple areas.

[0052] Casein peptide

[0053] In one embodiment, the method includes administering to a lactating mammal at least one milk-derived protein. In one embodiment, the milk-derived protein comprises casein or a casein-derived peptide. In one embodiment, the milk-derived protein further comprises β-lactoglobulin, α-lactalbumin, serum albumin, immunoglobulin G1 (IgG1), immunoglobulin G2 (IgG2), immunoglobulin A7 (IgA), immunoglobulin M (IgM), secretory component (SC), lactoferrin (LF), or any combination thereof.

[0054] Casein is a protein in the milk of non-human mammals and is also present in the milk of human mammals. It is known to include the subgroups αS1, αS2, β, and κ. Casein is defined according to the amino acid sequence of each of the subgroups αS1, αS2, β, and κ. In the context of the present disclosure, when referring to casein, it should be understood to also include acid casein, salts of casein, phosphoproteins, and rennet casein.

[0055] As used herein, the term "protein" refers to amino acid residues linked by peptide bonds. Protein sequences are typically 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 both natural and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that act in a manner similar to natural amino acids. Amino acids herein may be referred to by their commonly known three-letter symbols or by the single-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission.

[0056] Casein-derived peptides can be a single peptide or a mixture of different peptides, which can independently be selected from naturally occurring peptides, semi-synthetic peptides, synthetic peptides, or recombinant peptides. It should be further noted that the peptides according to the present disclosure can be produced synthetically, or by recombinant DNA technology, or by any other technique. Methods for producing peptides are well known in the art.

[0057] In some embodiments, the casein-derived peptide can comprise casein breakdown products that occur when casein is cleaved into peptide fragments by an enzyme or an acid (also referred to in the art by the term "casein hydrolysate"). Casein hydrolysate should be understood as the hydrolyzed form of casein (protein). Casein hydrolysates include, for example, the active β-, αS1-, αS2-, κ-casein-derived peptides known to those skilled in the art. In some embodiments, the casein-derived peptide is or comprises a casein hydrolysate.

[0058] In some embodiments, the casein-derived peptide comprises a natural peptide, a synthetic peptide, a semi-synthetic peptide, or any combination thereof. In another embodiment, the casein-derived peptide comprises a natural peptide. In another embodiment, the casein-derived peptide comprises a synthetic peptide. In another embodiment, the casein-derived peptide comprises a semi-synthetic peptide. In another embodiment, the casein-derived peptide comprises a combination of a natural peptide, a synthetic peptide, and a semi-synthetic peptide.

[0059] Native casein-derived peptides are typically obtained after enzymatic hydrolysis, and the enzyme can be any mammalian peptidase, such as, but not limited to, plasmin, pancreatin, trypsin, chymotrypsin, neutral protease, alkaline protease, pepsin, carboxypeptidase, cathepsin, as well as plant peptidases, such as, but not limited to, papain, bromelain, and enzymes from microbial sources. For example, naturally occurring casein-derived peptides may be the result of enzymatic activity (such as plasmin) on casein subunits β-casein, αs1- and αs2-casein, or κ-casein. In some embodiments, a casein hydrolysate is obtained by cleaving casein with trypsin.

[0060] Synthetic peptides can be obtained by any method known in the field of peptide synthesis, including chemical synthesis and recombinant DNA technology. For example, the peptides can be synthesized by using standard solid-phase techniques.

[0061] In one embodiment, the synthetic peptide is a recombinant peptide.

[0062] Semi-synthetic casein-derived peptides can be obtained by chemical hydrolysis of casein, for example, by prolonged boiling in strong acid (acid-HVP) or strong base, or by using chemical reagents such as cyanogen bromide (CNBr). The casein-derived peptides can also be obtained by molecular engineering, for example, using recombinant DNA, among the molecular techniques known in the art. In such an embodiment, the casein-derived peptide is a recombinant peptide.

[0063] In one embodiment, the recombinant peptide is produced by fermentation, tissue culture, or a combination thereof. In another embodiment, the recombinant peptide is produced by fermentation. In another embodiment, the recombinant peptide is produced by tissue culture. In another embodiment, the recombinant peptide is produced by a combination of fermentation and tissue culture.

[0064] In one embodiment, the tissue culture contains bovine mammary tissue.

[0065] 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 another embodiment, the casein-derived peptide comprises one or more fragments of αS2-casein. In another embodiment, the casein-derived peptide comprises one or more fragments of κ-casein. In another embodiment, the casein-derived peptide comprises one or more fragments of a combination of β-casein, αS1-casein, αS2-casein, and κ-casein.

[0066] In one embodiment, the casein-derived peptide further comprises amino acids of different lengths.

[0067] In one embodiment, the casein-derived peptide comprises a casein hydrolyzate.

[0068] In one embodiment, the casein-derived peptide comprises a phosphopeptide.

[0069] As used herein, the term "phosphopeptide" refers to a phosphorylated peptide in the form of a conjugated peptide, wherein the non-peptide moiety is a residue of phosphoric acid. The expression "phosphopeptide" or "phosphoserine" refers to conjugated serine, wherein the non-peptide moiety is a residue of phosphoric acid.

[0070] In some embodiments, the casein-derived peptide is a single peptide or a mixture of phosphopeptides, i.e., it contains a single phosphorus group or is a phosphorus-rich peptide. In some embodiments, the casein-derived peptide is any casein-derived peptide (casein phosphopeptide, CPP) rich in phosphoserine, phosphotyrosine, phosphothreonine, and / or phosphohistidine and a monovalent cation caseinate phosphate, such as sodium, potassium, calcium, or ammonium caseinate phosphate.

[0071] In some embodiments, the casein-derived peptide is a phosphorus-peptide.

[0072] The phosphorus-peptide can be a genetically engineered casein-derived peptide and a peptide mimetic of a casein-derived peptide. For example, phosphorylation of an amino acid (such as at least one serine residue) can be carried out by any method known in the art. The term "casein-derived peptide" also includes peptide fragments or peptide mimetic products obtained from or corresponding to one or more portions of casein. Peptide mimetic peptides can be, for example, peptoids or semi-peptoids, which are peptide analogs having, for example, modifications such as, but not limited to, cyclization, N-terminal modification, C-terminal modification, peptide bond modification, including but not limited to CH2-NH, CH2-S, CH2-S-O, O-C-NH, CH2-O, CH2-CH2, S-C-NH, CH-CH or CF-CH, backbone modification, and residue modification.

[0073] As used herein, the term "casein-derived peptide" further includes any derivative, analog, variant, or homolog of any peptide. The term "derivative" is used to define an amino acid sequence (peptide) in which any insertion, deletion, substitution, and modification of the amino acid sequence (peptide) do not change the activity of the original peptide. The term "derivative" also refers to its homologs, variants, and analogs, as well as covalent modifications of the polypeptides prepared according to the present invention.

[0074] In some embodiments, when two sequences are optimally aligned, 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%, or even 99% identical (in sequence) to the naturally occurring casein-derived peptides. In addition, any non-naturally occurring casein-derived peptide to be used in accordance with the present disclosure may retain at least a portion of the biological activity of the naturally occurring casein.

[0075] The present disclosure also includes homologs of the casein-derived peptides. The term "homolog" is used to define an amino acid sequence (peptide) that maintains a minimum homology with the amino acid sequence defined by the present invention, e.g., having at least about 65%, at least about 75%, at least about 85%, or at least about 95% total sequence homology with the amino acid sequence of any peptide (e.g., a specific sequence) structurally defined as above.

[0076] In some embodiments, the casein-derived peptides may also include chemical modifications of the naturally occurring peptides, such as where one or more amino acids are deleted, substituted, or modified, e.g., by removing a side group, substituting a side group, or introducing a chemical group. Without limitation, the chemical modifications may include acetylation, acylation, amidation, ADP-ribosylation, glycosylation, GPI anchor formation, covalent attachment of a lipid or lipid derivative, methylation, myristoylation, pegylation, prenylation, phosphorylation, ubiquitination, or any similar process. When referring to one amino acid sequence being replaced by another, such replacement is likely to be a conservative substitution. For example, one or more amino acid residues in the casein sequence are replaced by another amino acid having a similar polarity or charge. For example, non-polar (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 the desired (casein-like) biological activity of the resulting casein-derived peptide analog is not significantly altered.

[0077] The casein-derived peptides according to the present disclosure are characterized by a molecular weight between about an average of 100 and an average of 10,000 daltons (e.g., between 2 and 100 amino acids), sometimes between about an average of 100 and an average of 7,000 daltons, and sometimes between an average of 1,000 and an average of 5,000 daltons.

[0078] The casein-derived peptides according to the present disclosure are characterized by having a length of 2 to 200, 2 to 100 amino acids, sometimes between 4 and 40 amino acids, sometimes between 4 and 30 amino acids, sometimes between 4 and 10 amino acids, and sometimes between 10 and 50 amino acids.

[0079] In some embodiments, the casein-derived peptide comprises an amino acid sequence selected from SEQ ID NO.1 - SEQ ID NO.26.

[0080] In one embodiment, the phosphopeptide comprises the amino acid sequence represented as Ser-Ser-Ser-Glu (SEQ ID NO:1), wherein 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)).

[0081] In another embodiment, the phosphopeptide comprises the amino acid sequence represented as Ser-Ser-Ser-Glu-Glu (SEQ ID NO:2), wherein at least one Ser residue, at least two Ser residues, or three Ser residues are phosphorylated.

[0082] In another embodiment, the phosphopeptide comprises the amino acid sequence represented as Ser(p)-Ser(p)-Ser(p)-Glu-Glu (SEQ ID NO:3).

[0083] In another embodiment, the phosphopeptide comprises the amino acid sequence represented as RELEELNVPGEIVES(p)LS(p)S(p)S(p)EESITR (SEQ ID NO:4). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as QMEAESIS(p)S(p)S(p)EEIVPDSVEQK (SEQ ID NO:5). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as KNTMEHVS(p)S(p)S(p)EESIISNETYK (SEQ ID NO:6). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as KVNELSKNIGS(p)ES(p)TEDQ (SEQ ID NO:7). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as PTLNREQLS(p)TS(p)EENS KKTVD (SEQ ID NO:8). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as ELEELNVPGEIVES(p)LS(p)S(p)S(p)EESITR (SEQ ID NO:9). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as RELEELNVPGEIES(p)LS(p)S(p)S(p)EESITR (SEQ ID NO:10). In another embodiment, the phosphopeptide comprises the 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 the amino acid sequence represented as KNTME HVS(p)S(p)S(p)EESIIS(p)QETYK (SEQ ID NO:12). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as KVNELSKDIGS(p)ES(p)TEDQ (SEQ ID NO:13). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as ESIIS(p)QETYKQEKNMAINPSKENLCSTFCKEVVRNANEEETSIGS(p)S(p)S(p)EES(p)AEVATEEVKITVDDKHYQKALNEINQFYQKFPGYLQYLYQGPIVLNPWNQVLR NAVPITPTLNREQLS(p)TS(p)EENSKKTVN (SEQ ID NO:14). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as ELEELNVPGEIES(p)LS(p)S(p)S(p)EESITR (SEQ ID NO:15).

[0084] In another embodiment, the phosphopeptide comprises the sequence represented as X 1(n) -Ser(P)-Ser(P)-Ser(P)-X 2(m) (SEQ ID NO:16), where at least one of X1 and X2 is independently selected from positively charged amino acids, and where each of n and m is independently selected from 0, 1, and 2.

[0085] In some embodiments, the positively charged amino acid is selected from 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.

[0086] According to some embodiments, the formula of SEQ ID NO:16 further comprises a capping group (also referred to herein as a protecting group) located at the C-terminus. In some embodiments, the carboxyl group at the C-terminus of the peptide is protected by a protecting group. The protecting group is selected from, but not limited to, an amide (i.e., the hydroxyl group at the C-terminus is replaced by a primary amine (NH2), a secondary amine, or a tertiary amine) or an ester (i.e., the hydroxyl group at the C-terminus is replaced by an ester). According to some embodiments, the capping group is selected from amides and esters. According to some embodiments, the capping group is an amide.

[0087] In another embodiment, the phosphopeptide comprises the amino acid sequence represented as Lys-Lys-Ser(P)-Ser(P)-Ser(P) (SEQ ID NO:17). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as Lys-Lys-Ser(P)-Ser(P)-Ser(P)-Lys (SEQ ID NO:18). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as Lys-Lys-Ser(P)-Ser(P)-Ser(P)-Lys-Lys (SEQ ID NO:19). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as Lys-Ser(P)-Ser(P)-Ser(P)-Lys-Lys (SEQ ID NO:20). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as Lys-Ser(P)-Ser(P)-Ser(P)-Lys (SEQ ID NO:21). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as Lys-Ser(P)-Ser(P)-Ser(P) (SEQ ID NO:22). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as Ser(P)-Ser(P)-Ser(P)-Lys-Lys (SEQ ID NO:23). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as Ser(P)-Ser(P)-Ser(P)-Lys (SEQ ID NO:24). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as Lys-Lys-Ser(p)-Ser(p)-Ser(p)-NH2 (SEQ ID NO:25). In another embodiment, the phosphopeptide comprises the amino acid sequence represented as RELEELNVPGEIVES(p)LS(p)S(p)S(p)EESITRINK (SEQ ID NO:26).

[0088] The casein-derived peptides according to the present invention may comprise residues in the "L" and "D" forms. Although the amino acid residues of the peptide sequences listed in SEQ ID NO: 1-26 are all in the "L" isomer form, residues in the "D" isomer form may replace any L-amino acid residue, provided that the resulting peptide analog retains at least part 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 stability of the peptide against proteolytic degradation.

[0089] In some embodiments, the composition is free of antimicrobial agents and comprises an acceptable carrier.

[0090] In one embodiment, milk-derived proteins are measured by UV in the range of 204 to 220 nm.

[0091] Dose and Administration

[0092] In some embodiments, the methods of the present disclosure include administering milk-derived proteins between 10 ng / ml and 500 mg / ml. In one embodiment, the methods of the present disclosure include administering milk-derived proteins between 1 mg / ml and 500 mg / ml. In another embodiment, the methods of the present disclosure include administering milk-derived proteins between 10 mg / ml and 450 mg / ml. In another embodiment, the methods of the present disclosure include administering milk-derived proteins between 50 mg / ml and 400 mg / ml. In another embodiment, the methods of the present disclosure include administering milk-derived proteins between 50 mg / ml and 70 mg / ml. In another embodiment, the methods of the present disclosure include administering milk-derived proteins between 100 mg / ml and 350 mg / ml. In another embodiment, the methods of the present disclosure include administering milk-derived proteins between 150 mg / ml and 300 mg / ml. In another embodiment, the methods of the present disclosure include administering milk-derived proteins between 200 mg / ml and 250 mg / ml. In another embodiment, the methods of the present disclosure include administering milk-derived proteins between 5 mg / ml and 30 mg / ml.

[0093] In some embodiments, the methods of the present disclosure include intramammary infusion to a single teat or multiple teats. In one embodiment, the methods of the present disclosure include intramammary infusion to a single teat. In another embodiment, the methods of the present disclosure include intramammary infusion to multiple teats.

[0094] In some embodiments, the methods of the present disclosure include one to eight administrations. In one embodiment, the methods of the present disclosure include one administration. In one embodiment, the methods of the present disclosure include two administrations. In one embodiment, the methods of the present disclosure include three administrations. In one embodiment, the methods of the present disclosure include four administrations. In one embodiment, the methods of the present disclosure include five administrations. In one embodiment, the methods of the present disclosure include six administrations. In one embodiment, the methods of the present disclosure include seven administrations. In one embodiment, the methods of the present disclosure include eight administrations.

[0095] In some embodiments, the administration of the present disclosure includes an interval of from about 1 hour to about 72 hours. In one embodiment, the administration includes an interval of about 1 hour. In one embodiment, the administration includes an interval of about 4 hours. In one embodiment, the administration includes an interval of about 5 hours. In one embodiment, the administration includes an interval of about 8 hours. In one embodiment, the administration includes an interval of about 10 hours. In one embodiment, the administration includes an interval of about 12 hours. In one embodiment, the administration includes an interval of about 15 hours. In one embodiment, the administration includes an interval of about 16 hours. In one embodiment, the administration includes an interval of about 20 hours. In one embodiment, the administration includes an interval of about 24 hours. In one embodiment, the administration includes an interval of about 25 hours. In one embodiment, the administration includes an interval of about 28 hours. In one embodiment, the administration includes an interval of about 30 hours. In one embodiment, the administration includes an interval of about 35 hours. In one embodiment, the administration includes an interval of about 36 hours. In one embodiment, the administration includes an interval of about 40 hours. In one embodiment, the administration includes an interval of about 45 hours. In one embodiment, the administration includes an interval of about 50 hours. In one embodiment, the administration includes an interval of about 55 hours. In one embodiment, the administration includes an interval of about 60 hours. In one embodiment, the administration includes an interval of about 65 hours. In one embodiment, the administration includes an interval of about 70 hours. In one embodiment, the administration includes an interval of about 72 hours.

[0096] In some embodiments, the administration of the present disclosure includes an immediate administration of a double dose.

[0097] In some embodiments of the method of the present disclosure, the lactating mammal continues to produce and milk from a single or multiple untreated mammary regions. In one embodiment, the lactating mammal continues to produce and milk from a single untreated mammary region. In another embodiment, the lactating mammal continues to produce and milk from multiple untreated mammary regions.

[0098] In some embodiments, the milk is substantially free of residues. In one embodiment, the residues include antibiotic residues.

[0099] In some embodiments, the milk can be used as raw milk for dairy production, for breastfeeding, or any combination thereof. In one embodiment, the milk can be used as raw milk. In another embodiment, the milk can be used for dairy production. In another embodiment, the milk can be used for breastfeeding. In another embodiment, the milk can be used as raw milk for dairy production and for breastfeeding.

[0100] In some embodiments, the dairy products include milk, whey, yogurt, cheese, cream, butter, milk beverages with high protein, or combinations thereof. In one embodiment, the dairy product contains milk. In another embodiment, the dairy product contains whey. In another embodiment, the dairy product contains yogurt. In another embodiment, the dairy product contains cheese.

[0101] The term "casein" as used herein generally refers to the related family of proteins (αS1, αS2, β, κ) typically found in mammalian milk.

[0102] The term "treatment" relates to the improvement of at least one undesirable manifestation of a disease, such as an increase in the disease-free period, a decrease in the acute disease period (in terms of time and severity), a reduction in disease severity, an improvement in quality of life, an improvement in comfort and well-being, a reduction in mortality, a reduction in the disease progression rate, and prophylactic treatment before the occurrence of the disease. More specifically, the term "treatment or prevention" as used herein refers to all the positive therapeutic effects administered to a subject, including inhibiting, reducing, alleviating, and relieving a disorder or any related condition and disease, symptom, or undesirable side effect or related disorder. It should be understood that the term "reduce" or as referred to herein involves delaying, inhibiting, or reducing a process by about 1% to 99.9%, about 1% to about 5%, about 5% to 10%, about 10% to 15%, about 15% to 20%, about 20% to 25%, about 25% to 30%, about 30% to 35%, about 35% to 40%, about 40% to 45%, about 45% to 50%, about 50% to 55%, about 55% to 60%, about 60% to 65%, about 65% to 70%, about 75% to 80%, about 80% to 85%, about 85% to 90%, about 90% to 95%, about 95% to 99%, or about 99% to 99.9%.

[0103] The term "about" as used herein means a value that can deviate up to 1%, more specifically 5%, more specifically 10%, more specifically 15%, and in some cases up to 20% higher or lower than the indicated value. The deviation range includes integer values and, if applicable, also non-integer values, constituting a continuous range. As used herein, the term "about" means ±10%.

[0104] As used herein, the term "average molecular weight" refers to the average plus or minus standard deviation of the molecular weight of a peptide or protein, as measured by methods known to those of skill in the art. Such methods include, for example, SDS-gel electrophoresis and size exclusion chromatography in an apparatus such as HPLC, where the sample is run against standards of known molecular weight.

[0105] Example

[0106] Example 1. Induce a short non-milking period in the infected teat, followed by resumption of milk production from the infected teat during the same lactation period.

[0107] The aim is to determine the cure of intramammary infection during lactation by intramammary administration of casein or casein-derived peptides, followed by resumption of milk production from the infected teat during the same lactation period.

[0108] Study design

[0109] A multi-center, clinically controlled field study will allocate lactating dairy cows that test positive for bacteriological testing of intramammary infection to two treatment groups (casein or casein-derived peptides and negative control (untreated)) at a ratio of 1:1. The study population (23 dairy cows) is always divided into two groups: primiparous and multiparous. The dairy cows are recruited from commercial dairy farms.

[0110] Dairy cows with positive bacteriological results for the same pathogen from two bacterial milk samples and an elevated regional somatic cell count (QSCC) > 200,000 cells / ml are eligible for inclusion in the study, with the last sample being from one pre-treatment (last milking) sample.

[0111] Dairy cows are ineligible for inclusion in the study if they have been in the herd for less than 3 months, have a lactation length of less than 30 days from the estimated dry-off date, have less than 3 functional mammary regions, have received any antimicrobial, hormonal, anti-inflammatory drugs within 10 days prior to inclusion in the study, or according to the opinion of the investigator.

[0112] According to standard farming practices during lactation, the treated animals are housed in their regular accommodation. The cows are milked regularly from the untreated quarters. During treatment and follow-up, the dairy cows are housed, fed, and managed according to standard farm practices.

[0113] Data on the animals participating in the study for statistical analysis include the following: mastitis history, date of birth, number of lactations, date of last calving, number of milking days on the day of treatment, daily milk production and SCC from pre-treatment and post-treatment until 35 days after resumption of milking, and previous lactation.

[0114] Descriptions of housing and management practices, feeding and rationing programs, number of dairy cows, and teat disinfection procedures were collected from each farm participating in the study.

[0115] Physical examinations of dairy cows include rectal temperature, pulse, and respiratory rate.

[0116] Safety assessment parameters are recorded from general clinical examinations and clinical observations, and the dairy cows' feed intake and water intake are informed by farm staff.

[0117] Milk samples for bacteriological testing are collected separately from all areas or targeted udder areas, as outlined in Table 1 "Research Activity Schedule" below. The milk samples for bacteriological testing are collected into sterile 30 ml tubes. The volume of milk collected is approximately 5 ml.

[0118] Before treatment, milk samples for SCC are collected once separately from all areas, as outlined in Table 1 - "Research Activity Schedule" below. The milk samples for SCC are collected into 55 ml tubes. The volume of milk collected is approximately 35 ml.

[0119] According to laboratory instructions, all milk samples for bacteriology and SCC are stored in appropriate packaging by couriers or researchers and transported to the laboratory.

[0120] The laboratory provided the laboratory results to the researcher.

[0121] Table 1 - Research Activity Schedule

[0122]

[0123] SCC = Somatic Cell Count.

[0124] All raw data collection and procedures related to data collection, monitoring, and quality control checks are carried out according to standard operating procedures. This includes information on the research product administration procedures. According to the random sample table, dairy cows meeting the research criteria are evenly allocated to one of two groups.

[0125] The treatment group (T2) and the control group (T1) are randomly selected. The treatment group (T2) is treated with the research product in the infected and eligible udder areas, where an intramammary infusion of casein or casein-derived peptides is performed once. The treatment is carried out after the milking procedure. Before treatment, the teats are thoroughly cleaned and disinfected before each infusion.

[0126] After intramammary administration, the treated areas of the dairy cows are not milked. After a seven-day milking interval, milking is restarted. The remaining areas (healthy areas) and the untreated control (T1) areas are milked regularly.

[0127] The treatment unit and the statistical unit are single areas. At least a total of 10 dairy cows are recruited (including dropouts) in each treatment group.

[0128] Results and conclusions

[0129] After two milk sample tests, 23 out of 42 recruited dairy cows were bacteriologically positive. No adverse events occurred in the treated cows with casein or casein-derived peptides or in the untreated cows one day after treatment. On the 7th day after treatment, milking in all treated areas resumed and returned to the normal milking process.

[0130] The most prevalent isolated pathogen included non-Staphylococcus aureus (NAS).

[0131] After treatment, the results of bacteriological culture tests conducted on the 7th and 14th days after treatment showed that 8 out of 10 cases receiving casein or casein-derived peptides were negative, compared with 3 out of 10 cases in the untreated control group (see Table 2 below).

[0132] Table 2

[0133]

[0134]

[0135] (8) The negative bacteriological results of the two tests are indicated in parentheses

[0136] For udder regions with only 2 bacteriologically positive results from the same microorganism, milking was resumed again after the non-milking period on the 6 ± 1 day after treatment with casein or casein-derived peptides. On the 7th, 14th, and 21st days after treatment, bacteriological tests were performed on milk samples. Success was achieved only when both tests performed with the same microorganism were bacteriologically negative. In parentheses, the number of bacteriological negative results (successful treatment) of the two tests performed.

[0137] For non-Staphylococcus aureus (NAS), the success cure rates (bacteriologically negative results after treatment) for those treated with casein or casein-derived peptides and the untreated control were 8 out of 16 and 0 out of 7, respectively.

[0138] Therefore, the high cure rate results of the present invention provide advantages for the management of lactating animals, as it is safe, increases comfort, has no discarded milk after treatment, and does not use antibacterial agents.

[0139] Example 2. Induce a short non-milking period in the infected teat, and then resume milk production from the infected teat during the same lactation period.

[0140] The purpose of this example was to determine whether the lactation function of a single udder region could resume after the induction of regression by intramammary administration of casein or casein-derived peptides without affecting or increasing milk quality and yield.

[0141] Study Design

[0142] The dairy cows assigned to Example 1 were participants in a current multicenter case-control study.

[0143] Twenty-three dairy cows participated in this study after obtaining two bacteriological positive test results from milk samples of separate udder regions. Sixteen (16) received intramammary administration of casein or casein-derived peptides, and seven (7) were untreated (controls).

[0144] The udder regions of dairy cows that received intramammary infusion of casein or casein-derived peptides after milking were not milked for 7 ± 1 consecutive days. After 6 ± 1 days, milking of the treated udder regions was resumed. Other untreated udder regions were milked normally according to farm management. Conversely, the control udder regions continued to be milked throughout the treatment period. Somatic cell counts were recorded from the same period, as outlined in Table 3 - "Study Activity Schedule".

[0145] Milk production of all participating dairy cows was recorded daily from 35 days before the assignment or treatment date in the study until 35 days after resumption of milking.

[0146] Table 3 - Study Activity Schedule

[0147]

[0148]

[0149] SCC = Somatic Cell Count.

[0150] Results and Conclusions

[0151] After treatment of dairy cows treated in separate udder regions and untreated controls, all dairy cows completed a period of six (6 ± 1) days. After treatment, no adverse events occurred in any of the cows during the examination on day 14 after treatment.

[0152] The average weekly milk production before the trial, during the trial, and during resumption of milking is presented in Table 4 below.

[0153] Table 4 - Milk Production (Kg) of Dairy Cows Receiving Casein or Casein-Derived Peptides During the Trial, Average Weekly (Daily) Milk Production Averaged in Kg / Day.

[0154]

[0155] During the weeks (average 5 weeks) prior to the trial, the average daily milk production was similar between the two groups, at 35.5 kg / day for the untreated control group and 35.6 kg / day for the treated study group. During the one-week trial (7 days), the average daily milk volume recorded was 34.2 kg / day [measuring 4 nipples] in the untreated control group and 29.3 kg / day [measuring 3 nipples] in the treated group. During the one-week trial (6 ± 1 days) and the 5 weeks following the trial, the average daily milk volume recorded was 35.0 kg / day in the untreated control group and 36.12 kg / day in the treated group.

[0156] During the month prior to the trial, the combined SCC (values in '000 cells / ml) as described in Table 5 below was high and comparable for both study groups (1943 for the untreated control group and 2399 for the treated group). Similar high combined SCC values were recorded from day -3 to day -1 prior to treatment (2449 for the untreated control group and 2609 for the treated group). During the first month following the trial, the combined SCC in the treated group decreased to an average of 275 cells / ml, while the untreated control group was higher and recorded at 1,815 cells / ml. This represents a particularly low average among those animals treated with casein or casein-derived peptides (Table 5).

[0157] Table 5 - Composite somatic cell counts (expressed in '000 cells / ml) of dairy cows receiving casein or casein-derived peptides during the trial.

[0158]

[0159] *Range values are indicated in parentheses.

[0160] The lactating treatment group showed an immediate reduction in combined SCC immediately after the post-treatment period following the resumption of the milking process.

[0161] Examples 1 and 2 demonstrate that treatment with casein or casein-derived peptides enables resumption of milking within 7 days after treatment and exhibits significant economic value by reducing somatic cells and increasing milk production.

[0162] In addition to the immediate improvement in milk quality that persists within one month after the trial, another important added value of the treatment is the high bacterial cure rate, with milk not being discarded due to antibiotic residues.

[0163] Although certain features of the invention have been illustrated and described herein, many modifications, alternatives, variations, and equivalents will now occur to those of ordinary skill in the art. Accordingly, it is to be understood that the appended claims are intended to cover all such modifications and changes that fall within the true spirit of the invention.

Claims

1. A method for treating intramammary infections in lactating mammals during lactation, the method comprising administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the cure period of the lactating mammal occurs during the same lactation period.

2. The method according to claim 1, wherein the cure period of the lactating mammal comprises 60 days or fewer days after treatment.

3. The method according to claims 1 and 2, wherein the cure period of the lactating mammal comprises about 14 days or fewer days after treatment.

4. The method according to claims 1-3, wherein the cure period of the lactating mammal comprises about 7 days or fewer days after treatment.

5. The method according to claims 1-4, wherein the cure period enables the infected teat to resume milk production during the same lactation period.

6. The method according to claims 1-4, wherein the cure period enables the uninfected teats to continue milk production.

7. The method according to claim 1, wherein the milk production of the lactating mammal after resuming milk production is equal to or higher than the milk production before treatment.

8. The method according to claim 7, wherein the milk production before treatment comprises the milk production for up to 14 days after treatment.

9. The method according to claims 1-8, wherein the milk-derived protein comprises casein or casein-derived peptides.

10. The method according to claim 9, wherein the casein-derived peptides comprise natural peptides, synthetic peptides, semi-synthetic peptides, or any combination thereof.

11. The method according to claim 10, wherein the synthetic peptide is a recombinant peptide.

12. The method according to claim 11, wherein the recombinant peptide is produced by fermentation, tissue culture, or a combination thereof.

13. The method according to claim 12, wherein the tissue culture comprises bovine mammary tissue.

14. The method according to claims 9-13, wherein the casein-derived peptides comprise one or more fragments of β-casein, αS1-casein, αS2-casein, κ-casein, and optimally further comprise amino acids of different lengths or any combination thereof.

15. The method according to claims 9-14, wherein the casein-derived peptides comprise casein hydrolysates.

16. The method according to claims 9-15, wherein the casein-derived peptides comprise phosphopeptides.

17. The method according to claim 16, wherein the phosphopeptides comprise an amino acid sequence selected from the group consisting of SEQ ID NO.1-SEQ ID NO.

26.

18. The method according to claims 1-17, wherein the composition does not contain antibacterial agents and comprises an acceptable carrier.

19. The method according to claims 1-18, which comprises administering the milk-derived protein at between 10 ng / ml and 500 mg / ml.

20. The method according to claims 1-19, wherein the administration comprises intramammary infusion to a single teat or multiple teats.

21. The method according to claims 1-20, wherein the treatment comprises one to eight administrations.

22. The method according to any one of claims 1-21, wherein the administration comprises an interval of from about 1 hour to about 72 hours.

23. The method according to any one of claims 1-22, wherein the lactating mammal continues to produce and be milked from a single or multiple untreated mammary regions.

24. The method according to claim 23, wherein the milk is substantially free of residues, and wherein the milk can be used as raw milk for dairy production, for breastfeeding, or any combination thereof.

25. The method according to claim 24, wherein the dairy products comprise milk, whey, yogurt, cheese, cream, butter, milk beverages with high protein, or combinations thereof.

26. A method for continuously milking a lactating mammal during lactation, wherein the lactating mammal has a subclinical disease of intramammary infection, the method comprising administering to the lactating mammal a composition comprising at least one milk-derived protein, wherein the milking is from a single or multiple untreated mammary regions.

27. The method according to claim 26, wherein the subclinical disease comprises an increase in somatic cell count in the milk, an increase in milk conductivity, a decrease in milk volume, or any combination thereof.

28. The method according to claim 27, wherein the increase in somatic cell count, the increase in milk conductivity, the decrease in milk volume, or any combination thereof is the result of an uninfected or infected mammary region.

29. The method according to claims 27-28, wherein the somatic cell count in the milk is about 100,000 cells / ml or more.

30. The method according to claim 28, wherein at 25 °C, the increase in milk conductivity is about 3.0 millisiemens (mS) or higher from single or multiple measurements taken at intervals of from several hours to several months.

31. The method according to claims 26-30, wherein the milk-derived protein comprises casein or casein-derived peptides.

32. The method according to claim 31, wherein the casein-derived peptides comprise natural peptides, synthetic peptides, semi-synthetic peptides, or any combination thereof.

33. The method according to claim 32, wherein the synthetic peptide is a recombinant peptide.

34. The method according to claim 33, wherein the recombinant peptide is produced by fermentation, tissue culture, or a combination thereof.

35. The method according to claim 34, wherein the tissue culture comprises bovine mammary tissue.

36. The method according to claims 31-35, wherein the casein-derived peptides comprise one or more fragments of β-casein, αS1-casein, αS2-casein, κ-casein, or any combination thereof.

37. The method according to claims 31-36, wherein the casein-derived peptides comprise casein hydrolysates.

38. The method according to claims 31-37, wherein the casein-derived peptides comprise phosphopeptides.

39. The method according to claim 38, wherein the phosphopeptides comprise an amino acid sequence selected from the group consisting of SEQ ID NO.1-SEQ ID NO.

26.

40. The method according to claims 26 - 39, wherein the composition is free of antimicrobial agents and comprises an acceptable carrier.

41. The method according to claims 26 - 40, which comprises administering the milk-derived protein at a level between 10 ng / ml and 500 mg / ml.

42. The method according to claims 26 - 41, wherein the administration comprises intramammary infusion to a single teat or multiple teats.

43. The method according to claims 26 - 42, wherein the treatment comprises one to eight administrations.

44. The method according to claims 26 - 43, wherein the interval between administrations is from about 1 hour to about 72 hours.

45. The method according to claims 26 - 44, wherein the milk can be used as raw milk, for dairy production, for breastfeeding, or any combination thereof.

46. The method according to claim 45, wherein the dairy product comprises milk, whey, yogurt, cheese, cream, butter, milk beverages with high protein, or any combination thereof.

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