Methods of selectively promoting animal welfare through modulation of microbiome

A method for improving the health of a group of production animals kept in a confined space, the method comprising increasing the ratio of peripheral serotonin:tryptophan in the digestive system of said group of animals by feeding said group of production animals one of more of the following feed additives:

• N-acetyl-muramidase, and protease, wherein the ratio of peripheral serotonin:tryptophan in the brain of said group of animals is increased for at least 20% higher than the ratio of peripheral serotonin:tryptophan in the digestive system of a control group of animals which are fed with the same diet except for said feed additives;
• preferably wherein the N-acetyl-muramidase is selected from the group consisting of:
• (a) a polypeptide having at least 80% sequence identity to any one of SEQ ID NOs:
• 1-71;
• (b) a variant of a polypeptide having any one of SEQ ID NOs:
• 1-71 comprising one or more amino acid substitutions (preferably conservative substitutions), and/or one or more amino acid deletions, and/or one or more amino acid insertions or any combination thereof in 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 positions;
• (c) a polypeptide comprising the polypeptide of (a) or (b) and a N-terminal and/or C-terminal extension of between 1 and 10 amino acids;
• and (d) a fragment of a polypeptide of (a) or (b) having muramidase activity and having at least 90% of the length of the mature polypeptide;
• and
• preferably wherein the protease is selected from the group consisting of:
• (a′) a polypeptide having a sequence identity of at least 70% to any one of SEQ ID NOs 72-76;
• (b′) a variant of any one of SEQ ID NOs:
• 72-76, wherein the variant has protease activity and comprises one or more substitutions, and/or one or more deletions, and/or one or more insertions or any combination thereof in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50 positions;
• (c′) a polypeptide comprising the polypeptide of (a′) or (b′) and a N-terminal and/or C-terminal His-tag and/or HQ-tag;
• (d′) a polypeptide comprising the polypeptide of (a′) or (b′) and a N-terminal and/or C-terminal extension of up to 10 amino acids, e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids;
• and (e′) a fragment of the polypeptide of (a′) or (b′) having protease activity and having at least 90% of the length of the mature polypeptide.

A method for improving the health of a group of production animals kept in a confined space, the method comprising decreasing the ratio of tryptamine:tryptophan in the digestive system of said group of animals by feeding said group of production animals one of more of the following feed additives:

• N-acetyl-muramidase, and protease, wherein the ratio of tryptamine:tryptophan in the digestive system of said group of animals is decreased for at least 20% lower than the ratio of tryptamine:tryptophan in the digestive system of a control group of animals which are fed with the same diet except for said feed additives;
• preferably wherein the N-acetyl-muramidase is selected from the group consisting of:
• (a) a polypeptide having at least 80% sequence identity to any one of SEQ ID NOs:
• 1-71;
• (b) a variant of a polypeptide having any one of SEQ ID NOs:
• 1-71 comprising one or more amino acid substitutions (preferably conservative substitutions), and/or one or more amino acid deletions, and/or one or more amino acid insertions or any combination thereof in 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 positions;
• (c) a polypeptide comprising the polypeptide of (a) or (b) and a N-terminal and/or C-terminal extension of between 1 and 10 amino acids;
• and (d) a fragment of a polypeptide of (a) or (b) having muramidase activity and having at least 90% of the length of the mature polypeptide;
• and
• preferably wherein the protease is selected from the group consisting of:
• (a′) a polypeptide having a sequence identity of at least 70% to any one of SEQ ID NOs 72-76;
• (b′) a variant of any one of SEQ ID NOs:
• 72-76, wherein the variant has protease activity and comprises one or more substitutions, and/or one or more deletions, and/or one or more insertions or any combination thereof in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50 positions;
• (c′) a polypeptide comprising the polypeptide of (a′) or (b′) and a N-terminal and/or C-terminal His-tag and/or HQ-tag;
• (d′) a polypeptide comprising the polypeptide of (a′) or (b′) and a N-terminal and/or C-terminal extension of up to 10 amino acids, e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids;
• and (e′) a fragment of the polypeptide of (a′) or (b′) having protease activity and having at least 90% of the length of the mature polypeptide.

A method for improving the health of a group of production animals kept in a confined space, the method comprising increasing the ratio of melatonin:tryptophan in the digestive system of said group of animals by feeding said group of production animals one of more of the following group of feed additives:

• N-acetyl-muramidase, and protease, wherein the ratio of melatonin:tryptophan in the digestive system of said group of animals is increased for at least 10% higher than the ratio of melatonin:tryptophan in the digestive system of a control group of animals which are fed with the same diet except for said group of feed additives;
• preferably wherein the N-acetyl-muramidase is selected from the group consisting of:
• (a) a polypeptide having at least 80% sequence identity to any one of SEQ ID NOs:
• 1-71;
• (b) a variant of a polypeptide having any one of SEQ ID NOs:
• 1-71 comprising one or more amino acid substitutions (preferably conservative substitutions), and/or one or more amino acid deletions, and/or one or more amino acid insertions or any combination thereof in 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 positions;
• (c) a polypeptide comprising the polypeptide of (a) or (b) and a N-terminal and/or C-terminal extension of between 1 and 10 amino acids;
• and (d) a fragment of a polypeptide of (a) or (b) having muramidase activity and having at least 90% of the length of the mature polypeptide;
• and
• preferably wherein the protease is selected from the group consisting of:
• (a′) a polypeptide having a sequence identity of at least 70% to any one of SEQ ID NOs 72-76;
• (b′) a variant of any one of SEQ ID NOs:
• 72-76, wherein the variant has protease activity and comprises one or more substitutions, and/or one or more deletions, and/or one or more insertions or any combination thereof in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50 positions;
• (c′) a polypeptide comprising the polypeptide of (a′) or (b′) and a N-terminal and/or C-terminal His-tag and/or HQ-tag;
• (d′) a polypeptide comprising the polypeptide of (a′) or (b′) and a N-terminal and/or C-terminal extension of up to 10 amino acids, e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids;
• and (e′) a fragment of the polypeptide of (a′) or (b′) having protease activity and having at least 90% of the length of the mature polypeptide.

Use of feed enzymes, in particular N-acetyl-muramidase, and/or protease, in a diet for feeding to a group of animals for

• a) improving the health of said group of production animals kept in a confined space, comprising increasing the ratio of kynurenine:tryptophan in the body of said group of animals, wherein the ratio of kynurenine:tryptophan in the digestive system of said group of animals is increased for at least 10% higher than the ratio of kynurenine:tryptophan in the body of a control group of animals which are fed with the same diet except for said feed additives;
• b) improving the health of said group of production animals kept in a confined space, comprising increasing the ratio of peripheral serotonin:tryptophan in the digestive system of said group of animals, wherein the ratio of peripheral serotonin:tryptophan in the brain of said group of animals is increased for at least 20% higher than the ratio of peripheral serotonin:tryptophan in the digestive system of a control group of animals which are fed with the same diet except for said feed additives;
• c) improving the health of said group of production animals kept in a confined space, comprising decreasing the ratio of tryptamine:tryptophan in the digestive system of said group of animals, wherein the ratio of tryptamine:tryptophan in the digestive system of said group of animals is decreased for at least 20% lower than the ratio of tryptamine:tryptophan in the digestive system of a control group of animals which are fed with the same diet except for said feed additives;
• and/or
• d) improving the health of said group of production animals kept in a confined space, comprising increasing the ratio of melatonin:tryptophan in the digestive system of said group of animals, wherein the ratio of melatonin:tryptophan in the digestive system of said group of animals is increased for at least 10% higher than the ratio of melatonin:tryptophan in the digestive system of a control group of animals which are fed with the same diet except for said group of feed additives.