Compositions and methods for modulating mammary disorders and conditions

Abstract

The present invention provides compositions and methods for treating intramammary infection in lactating mammals during lactation and providing a continuous milk production and milking in lactating mammals during lactation, wherein somatic cell count in milk, milk electrical conductivity or combination thereof are increased.

Description

COMPOSITIONS AND METHODS FOR MODULATING MAMMARY DISORDERS AND CONDITIONSSEQUENCE LISTING STATEMENT

[0001] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said 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

[0002] The casein protein comprises three fractions, a, P and K, according to their electrophoretic mobility. Casein hydrolysate is the hydrolyzed form of casein which includes, among others, the active beta-casein-derived peptide. It has been established that casein hydrolysate plays a role in immune responses against microbial and viral infections.

[0003] Intramammary infection (IMI) plays a decisive role in breastfeeding humans and in the dairy industry impacting animal health, causing difficulties in maintaining milk quality and production, and negatively impacting welfare and comfort thus causing considerable economic losses.

[0004] IMI is a costly disease and needs to be identified as soon as possible to reduce the negative effect on milk quality and quantity and to maximize the chance of cure and prevent spreading.

[0005] Mastitis, is a disease caused generally by IMI and caused by pathogens, mostly bacteria, but also yeast, fungi, or even algae. Mastitis can be clinical, with local (and in some cases general) clinical signs and milk abnormalities, or subclinical with production losses and lowered milk quality.

[0006] Antimicrobial treatment is used in order to keep bovine udder health, and economic aspects in balance. On the contrary, emergence and spread of antimicrobial resistance (AMR) is an urgent matter of public interest, and consequently, extensive antimicrobialusage (AMU) in human, and animal health and in production livestock is being criticized.

[0007] Hygiene and management measures during milk collection, are likely to reduce but not control the incidence of both clinical and subclinical mastitis. Currently, many dairy parlors are equipped with on-line computerized data acquisition systems designed to detect milk and dairy cows’ related parameters. Thus, there is a recognized need for, and would behighly advantageous to have a composition and method to manage IMI at early stage during mammals’ lactation.SUMMARY OF THE INVENTION

[0008] In some aspects, disclosed herein is a method for treating intramammary infection in lactating mammals during lactation, comprising administering to the lactating mammals a composition comprising at least one milk-derived protein, wherein the lactating mammal cure period occurs during the same lactation. In some related aspects, the lactating mammal cure period comprises about 60 or less days posttreatment. In some related aspects, the lactating mammal cure period comprises about 45 or less days posttreatment. In some related aspects, the lactating mammal cure period comprises about 14 or less days posttreatment. In some related aspects, the lactating mammal cure period comprises about 7 or less days posttreatment.

[0009] In some related aspects, the cure period enables re-initiation of milk production form the infected teat during the same lactation. In some further related aspects, the cure period enables continuation of milk production from the uninfected teats.

[0010] In some further aspects, disclosed herein is a method for continuous milk production and milking in lactating mammals during lactation, wherein the lactating mammal is in a sub-clinical disease or clinical disease of intramammary infection comprising administering to the lactating mammals a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or plurality of udder quarters.

[0011] In some further aspects, the sub-clinical disease comprises increased somatic cell count in milk, increased milk electrical conductivity, reduction in milk quantity or combination thereof. In some related aspects, the increased somatic cell count in milk, increased milk electrical conductivity, reduction in milk quantity or any combination thereof are a result of uninfected or infected udder quarters.

[0012] In some related aspects, the lactating mammals continue milk production and milking from untreated single or plurality of udder quarters. In some further related aspects, the milk is essentially free of residues and can be used as raw milk, for dairy production, for breastfeeding or any combination thereof .In some further related aspects, the dairy product comprises milk, whey, yogurt, cheese, cream, butter, milk drinks with high protein or combination thereof.

[0013] In some related aspects, milk-derived protein comprises a casein protein or casein derived peptide.

[0014] In some further related aspects, the casein derived peptide comprises natural peptide, synthetic peptide, semi-synthetic peptide, or any combination thereof .

[0015] In some further related aspects, the casein derived peptide comprises one or more fragments of P-casein, aS 1 -casein, aS2-casein andK-casein. In some further related aspects, the casein derived peptide further comprises amino acids with different lengths.

[0016] In some further related aspects, the casein derived peptide comprises a casein hydrolysate. In some further related aspects, the casein derived peptide comprises a phosphopeptide. In some further related aspects, the phosphopeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO. 1 - SEQ ID NO. 26.DETAILED DESCRIPTION OF THE PRESENT INVENTION

[0017] In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the invention. However, it will be understood by those skilled in the art 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.Method of treatment

[0018] In some embodiments, disclosed herein is a method for treating intramammary infection in lactating mammals during lactation, comprising administering to the lactating mammals a composition comprising at least one milk-derived protein, wherein the lactating mammal cure period occurs during the same lactation.

[0019] In some embodiments, disclosed herein is a composition comprising at least one milk-derived protein, for use for in treating intramammary infection in lactating mammals during lactation, wherein the lactating mammal cure period occurs during the same lactation.

[0020] A person of ordinary skilled in the art would appreciate the term “cure” as treat, reduce symptoms, alleviate, change, stop, improve a condition (e.g., a disease), symptoms of a condition, or affect a condition (or disease), or to prevent or delay the onset of symptoms, complications, biochemical parameters of the disease or otherwise stop or inhibit the further development of the disease, or its symptoms.

[0021] In one embodiment, the lactating mammal cure period comprises 60 or less days posttreatment. In another embodiment, the lactating mammal cure period comprises 60 days posttreatment. In one embodiment, the lactating mammal cure period comprises 55 or less days posttreatment. In another embodiment, the lactating mammal cure period comprises 55 days posttreatment. In one embodiment, the lactating mammal cure period comprises 50 or less days posttreatment. In another embodiment, the lactating mammal cure period comprises 50 days posttreatment. In another embodiment, the lactating mammal cure period comprises 45 or less days posttreatment. In one embodiment, the cure period comprises 45 days or less. In another embodiment, the cure period comprises 45 days. In another embodiment, the cure period comprises 40 days or less. In another embodiment, the cure period comprises 40 days. In another embodiment, the cure period comprises 30 days or less. In another embodiment, the cure period comprises 30 days. In another embodiment, the cure period comprises 21 days. In another embodiment, the cure period comprises 20 days. In another embodiment, the cure period comprises 18 days. In another embodiment, the cure period comprises 16 days. In another embodiment, the cure period comprises 14 days. In another embodiment, the cure period comprises 12 days. In another embodiment, the cure period comprises 10 days. In another embodiment, the cure period comprises 9 days. In another embodiment, the cure period comprises 8 days. In another embodiment, the cure period comprises 7 days. In another embodiment, the cure period comprises 6 days. In another embodiment, the cure period comprises 5 days. In another embodiment, the cure period comprises 4 days. In another embodiment, the cure period comprises 3 days. In another embodiment, the cure period comprises 2 days.

[0022] In some embodiments, the cure period enables re-initiation of milk production form the infected teat during the same lactation. In one embodiment, the re-initiation of milk production is within the same lactation period. In another embodiment, the re-initiation of milk production is within the subsequent lactation period. In another embodiment, the reinitiation of milk production is from the treated single or plurality of udder quarters.

[0023] In some embodiments, the re-initiation of milk production occurs within 60 days of treatment or less. In other embodiments the re-initiation of milk production occurs within 45 days or less. In additional embodiments the re-initiation of milk production occurs with 30 days or less. In additional embodiments the re-initiation of milk production occurs within 14 days or less. In other embodiments the re-initiation of milk production occurs with 7 daysor less. In additional embodiments the re-initiation of milk production occurs within 5 days or less. In other embodiments the re-initiation of milk production occurs within 3 days or less.

[0024] In one embodiment, the cure period enables continuation of milk production from the uninfected teats.

[0025] In one embodiment, the milk production quantity from the re-initiation of milk production is equal to or higher from milk production quantity before treatment. In another embodiment, the milk production quantity from the re-initiation of milk production is equal to milk production quantity before treatment. In another embodiment, the milk production quantity from the re-initiation of milk production is higher than milk production quantity before treatment. In another embodiment, the milk production quantity from the re-initiation of milk production is higher than the milk production quantity of lactating mammals treated with antibiotics.

[0026] In one embodiment, the milk production quantity from the re-initiation of milk production is equal to or higher from milk production quantity before treatment after 2-90 days of re-initiation of milk production. In another embodiment, the milk production quantity from the re-initiation of milk production is equal to or higher from milk production quantity before treatment after 2 days of re-initiation of milk production. In another embodiment, the milk production quantity from the re-initiation of milk production is equal to or higher from milk production quantity before treatment after 10 days of re-initiation of milk production. In another embodiment, the milk production quantity from the re-initiation of milk production is equal to or higher from milk production quantity before treatment after 30 days of re-initiation of milk production. In another embodiment, the milk production quantity from the re-initiation of milk production is equal to or higher from milk production quantity before treatment after 50 days of re-initiation of milk production. In another embodiment, the milk production quantity from the re-initiation of milk production is equal to or higher than milk production quantity before treatment after 70 days of re-initiation of milk production. In another embodiment, the milk production quantity from the re-initiation of milk production is equal to or higher from milk production quantity before treatment after 90 days of re-initiation of milk production.

[0027] In one embodiment, the milk production quantity from the re-initiation milk production is higher than milk production quantity before treatment by 0.5%-10%. Inanother embodiment, the milk production quantity from the re-initiation milk production is higher from milk production quantity before treatment by 2%. In another embodiment, the milk production quantity from the re-initiation milk production is higher from milk production quantity before treatment by 3%. In another embodiment, the milk production quantity from the re-initiation milk production is higher than milk production quantity before treatment by 4%. In another embodiment, the milk production quantity from the reinitiation milk production is higher than milk production quantity before treatment by 5%. In another embodiment, the milk production quantity from the re-initiation milk production is higher than milk production quantity before treatment by 6%. In another embodiment, the milk production quantity from the re-initiation milk production is higher than milk production quantity before treatment by 7%. In another embodiment, the milk production quantity from the re-initiation milk production is higher than milk production quantity before treatment by 8%. In another embodiment, the milk production quantity from the reinitiation milk production is higher than milk production quantity before treatment by 9%. In another embodiment, the milk production quantity from the re-initiation milk production is higher than milk production quantity before treatment by 10%.

[0028] In another embodiment, the milk production quantity from the re-initiation milk production is higher than milk production quantity before treatment by more than 10%.

[0029] In one embodiment, milk production quantity before treatment comprises milk production quantity up to 60 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 60 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity up to 30 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 30 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity up to 25 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 25 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity up to 20 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 20 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity up to 15 days after treatment. In another embodiment, milk production quantity beforetreatment comprises milk production quantity 15 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity up to 14 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 14 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 12 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 10 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 8 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 7 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 6 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 5 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 4 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 3 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 2 days after treatment. In another embodiment, milk production quantity before treatment comprises milk production quantity 1 day after treatment.

[0030] In some embodiments, disclosed herein is a method for continuous milk production and milking in lactating mammals during lactation, wherein the lactating mammal is in a state of clinical disease of intramammary infection comprising administering to the lactating mammals a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or plurality of udder quarters.

[0031] In some embodiments, disclosed herein is a method for continuous milk production and milking in lactating mammals during lactation, wherein the lactating mammal is in a sub-clinical disease of intramammary infection comprising administering to the lactating mammals a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or plurality of udder quarters.

[0032] A skilled in the art would appreciate the term “sub clinical disease” for intramammary infection where the disease process, is from the susceptibility time ofexposure to the asymptomatic onset of the process, comprising increased somatic cell count in milk, milk electrical conductivity, and reduction in milk quantity or combination thereof.

[0033] A skilled in the art would appreciate the term “intramammary infection (IMI)” as the presence of an infectious organism in the mammary gland. The terms IMI clinical mastitis and subclinical mastitis are used almost interchangeably. Intramammary infection (IMI), comprises a group of costly diseases affecting animals, including humans, worldwide.

[0034] In some embodiments, the sub-clinical disease comprises increased somatic cell count in milk, increased milk electrical conductivity, reduction in milk quantity or any combination thereof. In one embodiment, the sub-clinical disease comprises increased somatic cell count in milk. In another embodiment, the sub-clinical disease comprises increased milk electrical conductivity. In another embodiment, the sub-clinical disease comprises reduction in milk quantity. In another embodiment, the sub-clinical disease comprises increased somatic cell count in milk, increased milk electrical conductivity and reduction in milk quantity.

[0035] In one embodiment, the increased somatic cell count in milk, increased milk electrical conductivity, reduction in milk quantity or any combination thereof are a result of uninfected or infected udder quarters. In another embodiment, the increased somatic cell count in milk, is a result of uninfected or infected udder quarters. In another embodiment, the increased milk electrical conductivity, is a result of uninfected or infected udder quarters. In another embodiment, the reduction in milk quantity, is a result of uninfected or infected udder quarters. In another embodiment, the increased somatic cell count in milk, increased milk electrical conductivity and reduction in milk quantity are a result of uninfected or infected udder quarters.

[0036] In one embodiment, disclosed herein is a method for continuous milk production and milking in lactating mammals during lactation, wherein somatic cell count in milk is increased from one single quarter or a plurality of quarters, comprising administering to the lactating mammals a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or plurality of udder quarters.

[0037] In one embodiment, disclosed herein is a method for continuous milk production and milking in lactating mammals during lactation, wherein milk electrical conductivity is increased from one single quarter or a plurality of quarters, comprising administering to thelactating mammals a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or plurality of udder quarters.

[0038] In one embodiment, disclosed herein is a method for continuous milk production and milking in lactating mammals during lactation, wherein milk quantity is reduced from one single quarter or a plurality of quarters, comprising administering to the lactating mammals a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or plurality of udder quarters.

[0039] In some embodiments, disclosed herein is a method for continuous milk production and milking in lactating mammals during lactation, wherein somatic cell count in milk, milk electrical conductivity or combination thereof are increased and wherein milk quantity is reduced from one single quarter or a plurality of quarters, comprising administering to the lactating mammals a composition comprising at least one milk-derived protein, wherein the milking is from an untreated single or plurality of udder quarters.

[0040] In one embodiment, the somatic cell count in milk is about 100,000 cells per ml or over. In another embodiment, the increase in somatic cell count in milk is about 150,000 cells per ml or over. In another embodiment, the increase in somatic cell count in milk is about 180,000 cells per ml or over. In another embodiment, the increase in somatic cell count in milk is about 1 million cells per ml or over. In another embodiment, the increase in somatic cell count in milk is about 5 million cells per ml or over. In another embodiment, the increase in somatic cell count in milk is about 10 million cells per ml or over.

[0041] In some embodiments, disclosed herein is a method for continuous milk production and milking in lactating mammals during lactation, wherein somatic cell count in milk and milk electrical conductivity are increased from one single quarter or a plurality of quarters, comprising administering to the lactating mammals a composition comprising at least one milk-derived protein.

[0042] In some embodiments, disclosed herein is a method for continuous milk production and milking in lactating mammals during lactation, comprising administering to the lactating mammals a composition comprising at least one milk-derived protein, wherein the comfort during milking is increased.

[0043] As used herein, the term "comfort" refers to the prevention of suffering and increasing the presence of positive feelings, usually called comfort or pleasure, resulting from, inter alia, an increase lying periods, an increase in ruminating time, a decrease inmetabolic need, a decrease in udder pressure and / or teat leakage, decrease in incidence of mastitis and other diseases, and decrease in lameness effect due to high milk yield.

[0044] In some embodiments, disclosed herein is a method for continuous milk production and milking in lactating mammals during lactation, comprising administering to the lactating mammals a composition comprising at least one milk-derived protein, wherein the period of treatment is at any stage of lactation, including drying off to one or plurality of udder quarters.

[0045] A skilled artisan would understand the term “during lactation” in the dairy industry as the period of time between one calving and the next calving.

[0046] In one embodiment, the increase of milk electrical conductivity is about 3.0 milliSiemens (mS) or more at 25 °C, from a single or multiple measures during an interval of hour up to several months between each. In another embodiment, the increase of milk electrical conductivity is about 4 milliSiemens (mS) or more at 25 °C. In another embodiment, the increase of milk electrical conductivity is about 5 milliSiemens (mS) or more at 25 °C. In another embodiment, the increase of milk electrical conductivity is about 6 milliSiemens (mS) or more at 25 °C. In another embodiment, the increase of milk electrical conductivity is about 7 milliSiemens (mS) or more at 25 °C. In another embodiment, the increase of milk electrical conductivity is about 8 milliSiemens (mS) or more at 25 °C. In another embodiment, the increase of milk electrical conductivity is about 9 milliSiemens (mS) or more at 25 °C. In another embodiment, the increase of milk electrical conductivity is about 10 milliSiemens (mS) or more at 25 °C.

[0047] In one embodiment, disclosed herein is a method for continuous milk production and milking in lactating mammals during lactation, wherein bacteriology is positive in one single quarter or a plurality of quarters comprising administering to the lactating mammals a composition comprising at least one milk-derived protein.In one embodiment, the bacteriology is positive in a single or multiple measures. In one embodiment, the bacteriology is positive in one single quarter or a plurality of quarters.

[0048] In one embodiment, the positive bacteriology in one single quarter or a plurality of quarters is associated with reduction of milk. In one embodiment, the reduction of milk production is by at least 2% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters. In one embodiment, the reduction of milk production is by at least 10% compared to milk production from negative bacteriology in one singlequarter or a plurality of quarters. In one embodiment, the reduction of milk production is by 10-50% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters. In another embodiment, the reduction of milk production is by 10% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters. In another embodiment, the reduction of milk production is by 20% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters. In another embodiment, the reduction of milk production is by 23% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters. In another embodiment, the reduction of milk production is by 25% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters. In another embodiment, the reduction of milk production is by 30% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters. In another embodiment, the reduction of milk production is by 35% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters. In another embodiment, the reduction of milk production is by 40% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters. In another embodiment, the reduction of milk production is by 45% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters. In another embodiment, the reduction of milk production is by 50% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters. In another embodiment, the reduction of milk production is by more than 50% compared to milk production from negative bacteriology in one single quarter or a plurality of quarters.Casein peptides

[0049] In one embodiment, the method comprises administering to the lactating mammals at least one milk-derived protein. In one embodiment, the milk-derived protein comprises a casein protein or casein derived peptide. In one embodiment, the milk derived protein further comprises P-Lactoglobulin, a-Lactalbumin, Serum albumin, Immunoglobulin G1 (IgGl), Immunoglobulin G2 (IgG2), Immunoglobulin A7 (IgA), Immunoglobulin M(IgM), Secretory component (SC), Lactoferrin (LF) or any combination thereof.

[0050] Casein is a protein in non-human mammal’ s milk, also found in human mammal’ s milk known to include the subgroups aSl, aS2, p and K. Casein is defined according to the amino acid sequences of each of the subgroups aSl, aS2, p and K. In the context of thepresent disclosure, when referring to casein, it is to be understood as also including acid casein, salts of casein, phosphorous containing casein and rennet casein.

[0051] The term "protein" as used herein refers to amino acid residues, connected by peptide bonds. A protein sequence is generally reported from theN-terminal end containing free amino group to the C-terminal end containing free carboxyl group. Amino acids, as used herein, refer to naturally occurring and synthetic amino acids, as well as amino acid analogs, and amino acid mimetics, that function in a manner similar to the naturally occurring amino acids. Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission.

[0052] The casein-derived peptide may be a single peptide or a mixture of different peptides which may be independently selected from a naturally occurring peptide, a semi -synthetic peptide, a synthetic peptide or a recombinant peptide. It should be further noted that the peptides according to the present disclosure may be produced synthetically, or by recombinant DNA technology, or by any other technology. Methods for producing peptides are well known in the art.

[0053] In some embodiments, the casein-derived peptide may comprise a casein protein breakdown product which occurs when casein protein is cleaved by enzymes or acids to peptide fragments (also known in the art by the term "casein hydrolysate"). A casein hydrolysate is to be understood as the hydrolyzed form of casein (protein). Casein hydrolysate includes, for example, the active beta-, alpha S1-, alpha S2-, kappa-caseinderived peptide known to those versed in the art. In some embodiments, the casein-derived peptide is or comprises a casein hydrolysate.

[0054] In some embodiments, the casein derived peptide comprises natural peptide, synthetic peptide, 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 natural peptide, synthetic peptide and semi-synthetic peptide.

[0055] Natural casein-derived peptides are typically obtained following enzymatic hydrolysis, the enzyme may be any mammal peptidase, such as, without being limitedthereto, plasmin, pancreatin, trypsin, chymotrypsin, neutrase, alcalase, pepsin, carboxypeptidase, cathepsin, as well as plant peptidase such as, without being limited thereto, papain, bromelain, as well as enzymes from microorganism source. For example, a naturally occurring casein-derived peptide may be the result of an enzyme activity such as plasmin on casein subunits P-casein, asl- and as2-casein or K-casein. In some embodiments, a casein hydrolysate is obtained by cleavage of the casein protein with trypsin.

[0056] A synthetic peptide may be obtained by any methods known in the art of peptide synthesis including chemical synthesis and recombinant DNA technology. For example, the peptides may be synthesized by using standard solid phase techniques.

[0057] In one embodiment, the synthetic peptide is a recombinant peptide.

[0058] A semi-synthetic casein-derived peptide may be obtained by chemical hydrolysis of casein, e.g. by prolonged boiling in a strong acid (acid-HVP) or strong base or using a chemical agent such as Cyanogen bromide (CNBr). The casein -derived peptide may also be obtained by molecular engineering, e.g. using recombinant DNA, in molecular techniques known in the art. In such an embodiment, the casein-derived peptide is a recombinant peptide.

[0059] In one embodiment, the recombinant peptide is produced by fermentation, tissue culture or 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.

[0060] In one embodiment, the tissue culture comprises mammary gland bovine tissue .

[0061] In some embodiments, the casein derived peptide comprises one or more fragments of P-casein, aS 1 -casein, aS2-casein, K-casein or any combination thereof. In one embodiment, the casein derived peptide comprises one or more fragments of P-casein. In another embodiment, the casein derived peptide comprises one or more fragments of aSl- casein. In another embodiment, the casein derived peptide comprises one or more fragments of aS2-casein. In another embodiment, the casein derived peptide comprises one or more fragments of K-casein. In another embodiment, the casein derived peptide comprises one or more fragments of combination of P-casein, aS 1 -casein, aS2-casein and K-casein.

[0062] In one embodiment, the casein derived peptide further comprises amino acids with different lengths.

[0063] In one embodiment, the casein derived peptide comprises a casein hydrolysate .

[0064] In one embodiment, the casein derived peptide comprises a phosphopeptide .

[0065] As used herein, the term "phosphopeptide" designates a phosphorylated peptide in form of a conjugated peptide in which the non-peptide portion is a residue of phosphoric acid. The expression "phosphopeptide" or "phosphoserine" designates conjugated serine in which the non-peptide portion is a residue of phosphoric acid.

[0066] In some embodiments, the casein-derived peptide is a single peptide or mixture of a phosphopeptide, namely, which contains a single phosphorous group or is a phosphorus- enriched peptide. In some embodiments, the casein-derived peptide is any phosphoserine, phosphotyrosine, phosphothreonine, and / or phosphohystidine-enriched casein-derived peptides (casein phosphopeptide, CPP) and monovalent cation phosphocaseinates, such as sodium, potassium, calcium or ammonium phosphocaseinates.

[0067] In some embodiments, the casein-derived peptide is a phosphor-peptide.

[0068] The phosphor-peptide may be genetically engineered casein-derived peptides as well as peptidomimetics of casein-derived peptides. For example, phosphorylation of amino acids such as at least one serine residue may be performed by any method as is known in the art. The term "casein-derived peptide" also encompasses peptide fragments or peptidomimetic products obtained from or corresponding to one or more sections of casein protein. The peptidomimetic peptide may be for example a peptoid or a semipeptoid, which are peptide analogs, having, for example, modifications such as, but are not limited to, cyclization, N-terminus modification, C-terminus 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.

[0069] As used herein, the term “casein-derived peptide” further encompasses any derivatives, analogues, variants or homologues of any of the peptides. The term "derivative" is used to define amino acid sequences (peptide), with any insertions, deletions, substitutions and modifications to the amino acid sequences (peptide) that do not alter the activity of the original peptides. By the term “derivative” it is also referred to homologues, variants and analogues thereof, as well as covalent modifications of polypeptides made according to the present invention.

[0070] In some embodiments, the modified, synthetic, semi -synthetic or other types of analogs of the naturally occurring casein-derived peptides are in some embodiments at least75%, at times 85%, 90%, 95% and even 99% identical (in sequence) to a naturally occurring casein-derived peptide when the two sequences are optimally aligned. Further, any non- naturally occurring casein-derived peptide to be used in accordance with the present disclosure may retain at least part of the biological activity of the naturally occurring casein protein.

[0071] The present disclosure also encompasses homologues of the casein-derived peptide. The term "homologues” is used to define amino acid sequences (peptide) which maintain a minimal homology to the amino acid sequences defined by the invention, e.g. have at least about 65%, at least about 75%, at least about 85%, or at least about 95% overall sequence homology with the amino acid sequence of any of the peptide as structurally defined above, e.g. of a specified sequence.

[0072] In some embodiments, the casein-derived peptide may also include a chemical modification of a naturally occurring peptide, e.g. where one or more amino acids are deleted, substituted or modified, e.g. by removal of a side group, substitution of a side group or the introduction of a chemical group. Without being limited thereto, the chemical modification 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 replacement of an amino acid sequence by another, it is likely that the replacement is a conservative substitution. For example, one or more amino acid residues within a casein sequence is substituted by another amino acid of a similar polarity or charge. For example, the non-polar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan and methionine. The polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine. The positively charged (basic) amino acids include arginine, lysine and histidine. The negatively charged (acidic) amino acids include aspartic acid and glutamic acid. Nonetheless, non-conservative substitutions may also take place as long as it does not significantly change the desired (casein like) biological activity of the resulting caseinderived peptide analog.

[0073] A casein-derived peptide in accordance with the present disclosure is characterized by a molecular weight of between about an average 100 to an average 10,000 Dalton (e.g.between 2 to 100 amino acids) at times between about an average 100 to an average 7,000 Dalton and at times between an average 1,000 to an average 5,000 Daltons.

[0074] A casein-derived peptide in accordance with the disclosure is characterized by a length of from 2 to 200, from 2 to 100 amino acids, at times between 4 amino acids to 40 amino acids, at times from 4 amino acids to 30 amino acids, at times 4 amino acids to 10 amino acids, at times between 10 amino acids to 50 amino acids.

[0075] In some embodiments, the casein-derived peptide comprises an amino acid sequence selected form the group consisting of SEQ ID NO. 1 - SEQ ID NO. 26.

[0076] In one embodiment, the phosphopeptide comprises an amino acid sequence denoted 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 denoted herein as Ser(p) or S(p)).

[0077] In another embodiment, the phosphopeptide comprises an amino acid sequence denoted 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.

[0078] In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as Ser(p)-Ser(p)-Ser(p)-Glu-Glu (SEQ ID NO:3).

[0079] In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as RELEELNVPGEIVES(p)LS(p)S(p)S(p)EESITR (SEQ ID NON). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as QMEAESIS(p)S(p)S(p)EEIVPDSVEQK (SEQ ID NO:5). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted asKNTMEHVS(p)S(p)S(p)EESIISNETYK (SEQ ID NO:6). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted asKVNELSKNIGS(p)ES(p)TEDQ (SEQ ID NO:7). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted asPTLNREQLS(p)TS(p)EENSKKTVD (SEQ ID NO: 8). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted asELEELNVPGEIVES(p)LS(p)S(p)S(p)EESITR (SEQ ID NO:9). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted asRELEELNVPGEIES(p)LS(p)S(p)S(p)EESITR (SEQ ID NO: 10). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted asQMEAES(p)IS(p)S(p)S(p)EEIVPNS(p)VEQK (SEQ ID NO: 11). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as KNTMEHVS(p)S(p)S(p)EESIIS(p)QETYK (SEQ ID NO: 12). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted asKVNELSKDIGS(p)ES(p)TEDQ (SEQ ID NO: 13). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted asESIIS(p)QETYKQEKNMAINPSKENLCSTFCKEWRNANEEETSIGS(p)S(p)S(p)EES (p)AEVATEEVKITVDDKHYQKALNEINQFYQKFPGYLQYLYQGPIVLNPWNQVL RNAVPITPTLNREQLS(p)TS(p)EENSKKTVN (SEQ ID NO: 14). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as ELEELNVPGEIES(p)LS(p)S(p)S(p)EESITR(SEQ ID NO: 15).

[0080] In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as Xi(n)- Ser(P)-Ser(P)-Ser(P)-X2(m) (SEQ ID NO: 16), wherein at least one of Xi and X2 is independently selected from a positively charged amino acid and wherein each one of n and m is independently selected from 0, 1 and 2.

[0081] 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.

[0082] According to some embodiments, the formula of SEQ ID NO:16 further comprises a blocking group (also denoted herein as a protecting group) at the C-terminus. In some embodiments, the carboxyl group at the C terminus of the peptide is protected with 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 with a primary amine (NH2), secondary amine, or tertiary amine) or ester (i.e. the hydroxyl group at the C terminus is replaced with an ester). According to some embodiments, the blocking group is selected from the group consisting of amide and ester. According to some embodiments, the blocking group is amide.

[0083] In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as Lys-Lys-Ser(P)-Ser(P)-Ser(P) (SEQ ID NO: 17). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as Lys-Lys-Ser(P)-Ser(P)- Ser(P)-Lys (SEQ ID NO: 18). In another embodiment, the phosphopeptide comprises anamino acid sequence denoted as Lys-Lys-Ser(P)-Ser(P)-Ser(P)-Lys-Lys (SEQ ID NO: 19). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as Lys-Ser(P)- Ser(P)-Ser(P)-Lys-Lys (SEQ ID NO:20). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as Lys- Ser(P)-Ser(P)-Ser(P)- Lys (SEQ ID NO:21). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as Lys-Ser(P)-Ser(P)-Ser(P) (SEQ ID NO:22). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as Ser(P)-Ser(P)-Ser(P)- Lys-Lys (SEQ ID NO:23). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as Ser(P)-Ser(P)-Ser(P)-Lys (SEQ ID NO:24). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as Lys-Lys- Ser(p)-Ser(p)-Ser(p)-NH2 (SEQ ID NO:25). In another embodiment, the phosphopeptide comprises an amino acid sequence denoted as RELEELNVPGEIVES(p)LS(p)S(p)S(p)EESITRINK (SEQ ID NO:26).

[0084] The casein-derived peptide according to the invention may comprise “L” as well as “D” form residues. While the amino acid residues of the peptide sequences set forth in SEQ ID NOs: l-26 are all in the "L" isomeric form, residues in the "D" isomeric form can substitute any L-amino acid residue so long as the resulting peptide analog retains at least part of the biological activity of the corresponding “L” isomer. One reason for designing casein-derived peptides comprising at least one D-amino acid is to increase stability of the peptide to proteolytic degradation.

[0085] In some embodiments, the composition is free of antimicrobials and comprises an acceptable carrier .

[0086] In one embodiment, the milk derived protein is measured by UV at a range from 204 to 220 nm.Dosage and administration

[0087] In some embodiments, the methods of the present disclosure comprise administering between lOng / ml to 500mg / ml of the milk derived protein. In one embodiment, the methods of the present disclosure comprise administering between Img / ml to 500mg / ml of the milk derived protein. In another embodiment, the methods of the present disclosure comprise administering between lOmg / ml to 450mg / ml of the milk derived protein. In another embodiment, the methods of the present disclosure comprise administering between 50mg / ml to 400mg / ml of the milk derived protein. In anotherembodiment, the methods of the present disclosure comprise administering between 50mg / ml to 70mg / ml of the milk derived protein. In another embodiment, the methods of the present disclosure comprise administering between lOOmg / ml to 350mg / ml of the milk derived protein. In another embodiment, the methods of the present disclosure comprise administering between 150mg / ml to 300mg / ml of the milk derived protein. In another embodiment, the methods of the present disclosure comprise administering between 200mg / ml to 250mg / ml of the milk derived protein. In another embodiment, the methods of the present disclosure comprise administering between 5mg / ml to 30mg / ml of the milk derived protein.

[0088] In some embodiments, the methods of the present disclosure comprise intramammary infusion to a single teat or a plurality of teats. In one embodiment, the methods of the present disclosure comprise intramammary infusion to a single teat. In another embodiment, the methods of the present disclosure comprise intramammary infusion to a plurality of teats.

[0089] In some embodiments, the methods of the present disclosure comprise between one to eight administrations. In one embodiment, the methods of the present disclosure comprise one administration. In one embodiment, the methods of the present disclosure comprise two administrations. In one embodiment, the methods of the present disclosure comprise three administrations. In one embodiment, the methods of the present disclosure comprise four administrations. In one embodiment, the methods of the present disclosure comprise five administrations. In one embodiment, the methods of the present disclosure comprise six administrations. In one embodiment, the methods of the present disclosure comprise seven administrations. In one embodiment, the methods of the present disclosure comprise eight administrations.

[0090] In some embodiments, the administrations of the present disclosure comprise intervals of from about 1 hour to about 72 hours . In one embodiment, the administrations comprise intervals of about 1 hour. In one embodiment, the administrations comprise intervals of about 4 hours. In one embodiment, the administrations comprise intervals of about 5 hours. In one embodiment, the administrations comprise intervals of about 8 hours. In one embodiment, the administrations comprise intervals of about 10 hours. In one embodiment, the administrations comprise intervals of about 12 hours. In one embodiment, the administrations comprise intervals of about 15 hours. In one embodiment, theadministrations comprise intervals of about 16 hours, In one embodiment, the administrations comprise intervals of about 20 hours, In one embodiment, the administrations comprise intervals of about 24 hours, In one embodiment, the administrations comprise intervals of about 25 hours, In one embodiment, the administrations comprise intervals of about 28 hours, In one embodiment, the administrations comprise intervals of about 30 hours, In one embodiment, the administrations comprise intervals of about 35 hours, In one embodiment, the administrations comprise intervals of about 36 hours, In one embodiment, the administrations comprise intervals of about 40 hours, In one embodiment, the administrations comprise intervals of about 45 hours, In one embodiment, the administrations comprise intervals of about 50 hours, In one embodiment, the administrations comprise intervals of about 55 hours, In one embodiment, the administrations comprise intervals of about 60 hours, In one embodiment, the administrations comprise intervals of about 65 hours, In one embodiment, the administrations comprise intervals of about 70 hours. In one embodiment, the administrations comprise intervals of about 72 hours.

[0091] In some embodiments, the administrations of the present disclosure comprise immediate administrations of a double dose.

[0092] In some embodiments of the methods of the present disclosure, the lactating mammals continue milk production and milking from untreated single or plurality of udder quarters. In one embodiment, the lactating mammals continue milk production and milking from untreated single udder quarter. In another embodiment, the lactating mammals continue milk production and milking from untreated plurality of udder quarters.

[0093] In some embodiments, the milk is essentially free of residues. In one embodiment, the residues comprise antibiotic residues.

[0094] 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 a 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 a raw milk, for dairy production and for breastfeeding.

[0095] In some embodiments, the dairy product comprises milk, whey, yogurt, cheese, cream, butter, milk drinks with high protein or combination thereof . In one embodiment,the dairy product comprises milk. In another embodiment, the dairy product comprises whey. In another embodiment, the dairy product comprises yogurt. In another embodiment, the dairy product comprises cheese.

[0096] The term “casein” as used herein generally refers to a family of related proteins (aSl, aS2, P, K) commonly found in mammalian milk.

[0097] The term "treatment" concerns improvement of at least one undesired manifestation of the disease such as increase in disease free periods, decrease in acute disease periods (in time and severely), decrease in severity of the disease, improvement in life quality, improvement in comfort and welfare, decreased mortality, decrease in the rate of disease progression as well as prophylactic treatment before disease occurs. More specifically, the term “treatment or prevention” as used herein, refers to the complete range of therapeutically positive effects of administrating to a subject including inhibition, reduction, alleviation and relief from a disorder or any related condition and illness, symptoms or undesired side effects or related disorders. It should be appreciated that the term “reduction” or as referred to herein, relate to the retardation, restraining or reduction of a process by any one of 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%.

[0098] The term "about" as used herein indicates values that may 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 value referred to, the deviation range including integer values, and, if applicable, non-integer values as well, constituting a continuous range. As used herein the term "about" refers to ± 10 %.

[0099] As used herein, the term “average molecular weight” refers to the mean plus or minus standard deviation of the molecular weight of the peptide or protein as measured by a method known to a person skilled in the art. Such methods include, for example, SDS-gel electrophoresis and size exclusion chromatography in an apparatus such as HPLC, wherein the sample is run against Standards with known molecular weight.EXAMPLESExample 1. Inducing a short non-milking period in infected teats, followed by re-initi- ation of milk production from the infected teat during the same lactation.

[0100] The objective is to determine the cure of intramammary infection during lactating period, by intramammary administration of casein protein or casein-derived peptides followed by re-initiation of milk production from the infected teat during the same lactation. Study Design

[0101] A multi centered, clinical controlled field study, allocating lactating cows bacteriology test positive for intramammary infection into two treatment groups in a ratio of 1 : 1 (casein protein or casein-derived peptides and negative control (untreated)). The study population (23 dairy cows) was consistently divided between primiparous and multiparous. Dairy cows were enrolled from commercial dairy farms.

[0102] Dairy cows were eligible for inclusion in the study with positive bacteriology results for the same pathogens from two bacteriological milk samples in conjunction with elevated quarter somatic cell count (QSCC) > 200,000 cells / ml, the last from one pre-treatment (last milking) sample.

[0103] Dairy cows were not eligible for inclusion in the study if they were in the herd less than 3 months, lactating less than 30 days from estimated dry-off day, less than 3 functional udder quarters, any antimicrobial, hormone, an anti-inflammatory medication has been administrated within 10 days before inclusion in the study, or in the Investigator’s opinion. Animals after treatment were maintained in their regular housing according to standard farming practices during lactation. Cows were regularly milked from the untreated quarters. During treatment and follow-up periods dairy cows are housed, feed and managed as per standard farm practice.

[0104] Data of the animal joining the study, for statistical analysis was included as follows: history of mastitis, birth date, number of lactations, date of the last calving, days in milking at day of treatment, daily milk yield and SCC from pre-treatment and post-treatment up to 35 days after re-milking and previous lactations.

[0105] A description of the housing and management practice, feed and feeding regimen, number of dairy cows, and teat disinfection procedures were collected from each farm which participated in the study.

[0106] A physical examination of the cow included rectal temperature, pulse, respiratory rate.

[0107] Safety assessment parameters were registered from general clinical examination and clinical observation, and by farm staff informing of the cow on feed intake of the cow and water intake.

[0108] Milk samples for bacteriology tests were collected separately from all quarters or the target udder quarter as summarized in the below Table 1 ‘Study Schedule of Events’. Milk samples for bacteriology tests were collected into sterile 30 ml tubes. The volume of milk collected was approximately 5 ml.

[0109] Milk samples for SCC were collected separately from all quarters once before treatment as summarized in the below Table 1 - ‘Study Schedule of Events’. Milk samples, for SCC, were collected into 55 ml tubes. The volume of milk collected was approximately 35 ml.

[0110] All milk samples for bacteriology and SCC were stored and shipped by courier or study personnel in suitable packaging to the laboratory according to lab instructions.The laboratory provided laboratory results to the Investigator.Table 1 - Study Schedule of EventsSCC= somatic cell counts.

[0111] All raw data collection and procedures relating to data collection, monitoring and quality control checks were conducted in accordance with standard operative procedure. This includes information about the investigational product administration procedure. Eligible dairy cows for the study were equally allocated according to a random sample table into one of the 2 groups.

[0112] Treatment (T2) and controls (Tl) were randomly selected, the treatment group (T2) was treated with the investigational product in the infected and eligible udder quarter with one intramammary infusion of casein protein or casein-derived peptides. Treatment was carried out after post-milking procedure. Before treatment the teats were thoroughly cleaned and disinfected before each infusion.

[0113] Cow’s treated quarters were not milked following the intramammary administration. After seven days of milking intermission, milking was re-initiated. The rest of the quarters (healthy quarters) and the untreated control (Tl) quarter were milked regularly.

[0114] The treatment unit and the statistical unit was the individual quarter. At least a total of 10 cows per treatment group were enrolled (including drop-out).Results and Conclusion

[0115] Twenty -three of 42 dairy cows enrolled were bacteriology positive after 2 milk samples tests. No adverse events occurred to cows either following treatment with casein protein or casein-derived peptides or untreated up to day posttreatment. All treated quarters reinitiated and returned to normal milking process at day 7 posttreatment.

[0116] The most prevalent isolated pathogens constituted non-aureus staphylococci (NAS).

[0117] Following treatment, results from both bacteriology culture tests performed at days 7 and 14 post-treatment indicated negative for 8 out of 10 received casein protein or caseinderived peptides, compared to 3 out of 10 in untreated controls (below Table 2).Table 2(8) in parenthesis denotes negative bacteriology results from two tests

[0118] Only, to udder quarter with 2 bacteriology positive result from same microorganism, a period of non-milking was followed by re-milking at day 6 ±1 after treatment with casein protein or casein-derived peptides. Bacteriology tests from milk samples were performed on days 7, 14 and 21 posttreatment. Success was achieved only when the 2 tests were bacteriology negative with the same microorganism treated. In parenthesis, number of bacteriology negative results (which is treatment success) of both tests performed.

[0119] To Non-< / / / v / / .s staphylococci (NAS) the success cure (bacteriology negative results following treatment) was 8 out of 16, and 0 out of 7 to those treated with casein protein or casein-derived peptide, and untreated controls, respectively.

[0120] Thus, the present high cure results provide an advantage in management of lactating animals as it is safe, increases comfort, no discarded milk following treatment, and no use of antimicrobials.Example 2. Inducing a short non-milking period in infected teats, followed by re-initi- ation of milk production from the infected teat during the same lactation.

[0121] The objectives of the present example were to determine whether lactation function from an individual udder quarter could be reinitiated after induced involution by intramammary administration of casein protein or casein-derived peptides without affecting or increasing the milk quality and production.Study Design

[0122] Dairy cows allocated to Example 1 were participants in the current multicenter case- controlled study.

[0123] Twenty three dairy cows participated in the study after 2 bacteriologies positive test results from milk samples from a sole udder quarter. Sixteen (16) received intramammary administration of casein protein or casein-derived peptides, and seven (7) were untreated (controls).

[0124] Udder quarter of dairy cow receiving intramammary infusion of casein protein or casein-derived peptides post milking, was not milked during 7±1 consecutive days. Milking was reinstituted after 6±1 days of the treated udder quarter. The other untreated udder quarters were normally milked according to farm management. Conversely, control udder quarter continued being milked during the whole treatment period. Somatic cell counts were recoded from the same period as summarized in the Table 3 - ‘Study Schedule of Events’

[0125] Milk yield from all participating cows was recorded daily from 35 days before day of allocation or treatment in the study up to 35 days after re-initiation of milking.Table 3 - Study Schedule of EventsSCC= somatic cell counts.Results and Conclusions

[0126] All dairy cows finished the six (6±1) days period following treatment to a sole udder quarter treated cows and untreated controls. Following treatment, in the period to the examination on day 14 posttreatment, no adverse events occurred to all cows.Average weekly milk yield during the pretrial, trial and re-initiation of milking periods are presented in the below Table 4.Table 4 - Milk Yield (Kg) from Dairy Cows Receiving casein protein or casein derived peptides by Trial Periods, Weekly average (per day) Milk Yield average of Kg / Day.

[0127] Daily average of the milk yield between both groups during the pre-trial weeks (average of 5 weeks) was similar, 35.5 kg / day and 35.6 / day kg for untreated controls and treated study groups. Daily average milk during the week trial (7 days) recorded was 34.2 kg\day in the untreated control [measurement of 4 teats] and 29.3 kg / day in the treated group [measurement of 3 teats]. Daily average milk during the week trial (6±1 days) and the 5 weeks post-trial recorded was 35.0 kg\day in the untreated control and 36.12 kg / day in the treated group.

[0128] Composite SCC as described in the below table 5, during the pre-trial month (values in ‘000 cell / ml) were high and comparable for both study groups (1943 untreated control and 2399 treated,). Similar high composite SCC values were recorded on days -3 to -1 before treatment (2449 untreated control and 2609 treated). Composite SCC decreased to an average of 275 cells / ml during the first post-trial month in the treated group compared to untreated controls which were higher and recorded 1,815 cells / ml. Such represent particularlow average values in those animals treated with casein protein or casein-derivate peptides(Table 5)Table 5 - Composite Somatic Cell Counts from Dairy Cows Receiving Casein Protein or Casein-derivate Peptides by Trial Periods (expressed in ‘000 cells / ml).*in parenthesis denotes range values.

[0129] Treatment group during lactation indicated a decrease of composite SCC in the immediate posttreatment time after reinstating the milking process.

[0130] Examples 1 and 2 showed that treatment with casein protein or casein-derivate peptides enable a re-initiation of milking within 7 days post treatment and exhibited a significant economic value by decreasing somatic cell and increasing milk yield.

[0131] In addition to the immediate improvement in milk quality that persisted in the posttrial month, another important added value from the treatment was a high rate of bacterial cure, undischarged milk resulting from antibiotic residue.

[0132] While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will now occur to those of ordinary skill in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.

Claims

CLAIMSWhat is claimed is:

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

2. The method according to claim 1, wherein said lactating mammal cure period comprises 60 or less days posttreatment.

3. The method according to claims 1 and 2, wherein said lactating mammal cure period comprises about 14 or less days posttreatment.

4. The method according to claims 1-3, wherein said lactating mammal cure period comprises about 7 or less days posttreatment.

5. The method according to claims 1 -4, wherein said cure period enables re-initiation of milk production from the infected teats during the same lactation.

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

7. The method according to claim 1, wherein said lactating mammal’s milk production quantity after re-initiation milk production is equal to or higher than milk production quantity before treatment.

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

9. The method according to claims 1 -8, wherein said milk derived protein comprises a casein protein or casein derived peptide.

10. The method according to claim 9, wherein said casein derived peptide comprises natural peptide, synthetic peptide, semi -synthetic peptide, or any combination thereof.

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

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

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

14. The method according to claims 9-13, wherein said casein derived peptide comprises one or more fragments of P-casein, aS 1 -casein, aS2-casein, K-casein, optimally further comprises amino acids with different lengths or any combination thereof.

15. The method according to claims 9-14, wherein said casein derived peptide comprises a casein hydrolysate.

16. The method according to claims 9-15, wherein said casein derived peptide comprises a phosphopeptide.

17. The method according to claim 16, wherein said phosphopeptide comprises an amino acid sequence selected form the group consisting of SEQ ID NO. 1 - SEQ ID NO. 26.

18. The method according to claims 1-17, wherein said composition is free of antimicrobials and comprises an acceptable carrier.

19. The method according to claims 1-18, comprising administering between lOng / ml to 500mg / ml of said milk derived protein.

20. The method according to claims 1-19 wherein said administration comprises intramammary infusion to a single teat or a plurality of teats.

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

22. The method according to claims 1-21 wherein said administration comprises intervals of from about 1 hour to about 72 hours.

23. The method according to claims 1-22, wherein said lactating mammals continue milk production and milking from untreated single or plurality of udder quarters.

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

25. The method according to claim 24, wherein said dairy product comprises milk, whey, yogurt, cheese, cream, butter, milk drinks with high protein or combination thereof.

26. A method for continuous milk production and milking in lactating mammals during lactation, wherein the lactating mammal is in a sub-clinical disease of intramammary infection comprising administering to said lactating mammals a composition comprising at least one milk-derived protein, wherein said milking is from an untreated single or plurality of udder quarters.

27. The method according to claim 26, wherein said sub-clinical disease comprises increased somatic cell count in milk, increased milk electrical conductivity, reduction in milk quantity or any combination thereof.

28. A method according to claim 27, wherein said increase in somatic cell count, electrical conductivity in milk, reduction in milk quantity or any combination thereof are a result of uninfected or infected udder quarters.

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

30. The method according to claim 28, wherein said increase of milk electrical conductivity is about 3.0 milliSiemens (mS) or more at 25 °C, from a single or multiple measures during an interval of hour up to several months between each.

31. The method according to claims 26-30, wherein said milk derived protein comprises a casein protein or casein derived peptide.

32. The method according to claim 31, wherein said casein derived peptide comprises natural peptide, synthetic peptide, semi -synthetic peptide, or any combination thereof.

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

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

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

36. The method according to claims 31-35, wherein said casein derived peptide comprises one or more fragments of P-casein, aS 1 -casein, aS2-casein, K-casein or any combination thereof.

37. The method according to claims 31-36, wherein said casein derived peptide comprises a casein hydrolysate.

38. The method according to claims 31-37, wherein said casein derived peptide comprises a phosphopeptide.

39. The method according to claim 38, wherein said phosphopeptide comprises an amino acid sequence selected form the group consisting of SEQ ID NO. 1 - SEQ ID NO.

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

41. The method according to claims 26-40, comprising administering between lOng / ml to 500mg / ml of said milk derived protein.

42. The method according to claims 26-41 wherein said administration comprises intramammary infusion to a single teat or a plurality of teats.

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

44. The method according to claims 26-43 wherein said administration is in intervals of from about 1 hour to about 72 hours.

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

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

Images