Oligosaccharide formulations acting against respiratory diseases

Abstract

The present invention relates to oligosaccharide formulations for treating, alleviating, preventing and / or delaying respiratory diseases, preferably infectious bronchitis and / or their action.

Description

[0001] This invention relates to oligosaccharide preparations for treating, alleviating, preventing and / or delaying respiratory diseases, preferably infectious bronchitis and / or their effects.

[0002] Global demand for poultry products continues to rise, while farmers face ongoing financial pressures. Therefore, ensuring animal welfare, sustainability, and animal health, while simultaneously optimizing animal performance and product quality, is crucial in poultry farming.

[0003] Infectious diseases, particularly respiratory ones such as infectious bronchitis (IB) or Newcastle disease (ND), occur in all regions and are a major cause of economic losses, especially in poultry farming. Clinical symptoms in birds infected with IB or NDV include wheezing, coughing, sneezing, respiratory epithelial damage, mucus buildup, tracheal rales, conjunctivitis, sinusitis, dyspnea, and suffocation. Simultaneously, feed consumption and weight gain, egg production and egg quality decrease, and feed conversion ratio (FCR) increases. In unvaccinated populations, morbidity is typically 100%, and mortality can reach up to 60%, depending on the specific virus strain in the infected population.

[0004] As a current countermeasure in poultry farming, birds are routinely vaccinated against NDV and IBV, for example by spraying chicks with live attenuated IBV. However, IBV taxonomic units include several strains, and new IBV types and variants emerge from mutation and recombination events. Therefore, currently applied IBV vaccines only provide cross-protection against some of these variants, and even with vaccination, IB outbreaks cannot be completely prevented.

[0005] In view of the above-mentioned prior art, one object of the present invention is to provide means and methods for treating, alleviating, preventing and / or delaying respiratory diseases and effects related to and / or caused by said respiratory diseases.

[0006] Surprisingly, this objective is achieved by using oligosaccharide formulations to prevent and / or delay respiratory diseases, preferably infectious bronchitis, and / or the effects of said respiratory diseases, preferably infectious bronchitis (i.e., infectious bronchitis caused by IBV); wherein said oligosaccharide formulations comprise at least n oligosaccharide fractions (DP1 to DPn fractions) each having different degrees of polymerization selected from 1 to n, where n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0007] In another aspect, the present invention relates to the use of oligosaccharide formulations for improving the efficacy of vaccines against respiratory diseases, preferably infectious bronchitis; wherein the oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having different degrees of polymerization selected from 1 to n, wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0008] In another aspect, the present invention relates to the use of oligosaccharide formulations for reducing, preventing, and / or delaying mortality from respiratory diseases in poultry populations challenged by respiratory diseases, preferably said respiratory diseases being infectious bronchitis (particularly IB caused by IBV); said oligosaccharide formulations comprising at least n oligosaccharide fractions (DP1 to DPn fractions) each having a different degree of polymerization selected from 1 to n, where n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0009] In another aspect, the present invention relates to the use of oligosaccharide formulations for reducing, mitigating, preventing and / or delaying the incidence of respiratory diseases (particularly the use of such formulations in poultry populations challenged by or at risk of being challenged by respiratory diseases), preferably wherein the respiratory disease is infectious bronchitis (particularly IB caused by IBV); wherein the oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having a different degree of polymerization selected from 1 to n, wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0010] In another aspect, the present invention relates to oligosaccharide formulations for use in methods of treating, alleviating, preventing and / or delaying the effects of respiratory diseases, preferably infectious bronchitis (particularly IB caused by IBV) and / or said respiratory diseases; wherein the oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having different degrees of polymerization selected from 1 to n, where n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0011] In some embodiments, the present invention relates to oligosaccharide formulations for use in methods of treating, alleviating, preventing, and / or delaying the effects of respiratory diseases, preferably infectious bronchitis, and / or said respiratory diseases; wherein said oligosaccharide formulations comprise at least n oligosaccharide fractions (DP1 to DPn fractions) each having a different degree of polymerization selected from 1 to n, where n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry; and wherein said oligosaccharide formulations are administered to individuals (e.g., animals, such as birds, poultry, or chickens) who have received a vaccine against a respiratory disease.

[0012] In some implementations, the oligosaccharide preparation is administered to individuals (e.g., animals, such as birds, poultry, or chickens) who will receive a vaccine against a respiratory disease.

[0013] In some implementations, the oligosaccharide preparation is administered to individuals (e.g., animals, such as birds, poultry, or chickens) who have received at least a first vaccination against a respiratory disease and will receive at least a second vaccination against a respiratory disease.

[0014] In some embodiments, the oligosaccharide formulation of the present invention is included in the nutritional composition at a concentration greater than 40 ppm; for example, greater than 40 ppm, 45 ppm, 50 ppm, 60 ppm, 70 ppm, 80 ppm, 90 ppm, 100 ppm, 110 ppm, 120 ppm, 130 ppm, 140 ppm, 150 ppm, 160 ppm, 170 ppm, 180 ppm, 190 ppm, 200 ppm, 210 ppm, 220 ppm, 230 ppm, 240 ppm, 250 ppm, 260 ppm, 270 ppm, 280 ppm, 290 ppm, 300 ppm, 310 ppm, 320 ppm, 330 ppm, 340 ppm, 350 ppm, 360 ppm, 370 ppm, 380 ppm, 390 ppm, 400 ppm, 410 ppm. ppm, 420 ppm, 430 ppm, 440 ppm, 450 ppm, 460 ppm, 470 ppm, 480 ppm, 490 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm, 1000 ppm, etc.

[0015] In some embodiments, the oligosaccharide formulation of the present invention has n as at least 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. 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100.

[0016] In some embodiments, at least one fraction of the oligosaccharide formulation of the present invention comprises less than 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, or 2% of oligosaccharides containing dehydrated subunits, based on relative abundance; and / or each fraction of the oligosaccharide formulation of the present invention comprises greater than 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, 50%, 60%, 70%, or 80% of oligosaccharides containing dehydrated subunits, based on relative abundance.

[0017] In some embodiments, the weight-average molecular weight of the oligosaccharide formulations of the present invention is about 300 to 5000 g / mol, 500 to 5000 g / mol, 700 to 5000 g / mol, 500 to 2000 g / mol, 700 to 2000 g / mol, 700 to 1500 g / mol, 300 to 1500 g / mol, 300 to 2000 g / mol, 400 to 1300 g / mol, 400 to 1200 g / mol, 400 to 1100 g / mol, 500 to 1300 g / mol, 500 to 1200 g / mol, 500 to 1100 g / mol, 600 to 1300 g / mol, 600 to 1200 g / mol, or 600 to 1100 g / mol; and / or, the number-average molecular weight of the oligosaccharide formulations of the present invention is about 1000 to 2000 g / mol. g / mol, 1100 to 1900 g / mol, 1200 to 1800 g / mol, 1300 to 1700 g / mol, 1400 to 1600 g / mol or 1450 to 1550 g / mol.

[0018] In some embodiments, the relative abundance of oligosaccharides in each of the n fractions of the oligosaccharide formulation of the present invention decreases monotonically with its degree of polymerization.

[0019] In some embodiments, the average DP of the oligosaccharide formulation of the present invention is at least 2. In some embodiments, the average DP of the oligosaccharide formulation of the present invention is at least 3.

[0020] In some embodiments, the average DP of the oligosaccharide formulation of the present invention is less than 5.0.

[0021] In some embodiments, the average DP of the oligosaccharide formulation of the present invention is greater than or equal to 2, for example at least 3, or at least 4; and the average DP of the oligosaccharide formulation of the present invention is less than 5.0.

[0022] In some embodiments, the average DP of the oligosaccharide formulation of the present invention is between 1 and 5; for example, between 2 and 5, or between 2 and 4.94, or between 3 and 5, or between 3 and 3.94.

[0023] In a preferred embodiment, the average DP of the oligosaccharide formulation of the present invention is determined based on the number-average molecular weight distribution of the oligosaccharide formulation of the present invention.

[0024] A further feature of the present invention is the following item: Item 1. The use of an oligosaccharide formulation for the treatment, relief, prevention and / or delay of respiratory diseases, preferably infectious bronchitis (particularly infectious bronchitis caused by IBV) and / or its effects (i.e., the effects of said respiratory diseases, or the effects caused by said respiratory diseases, or the effects associated with said respiratory diseases); wherein said oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having different degrees of polymerization selected from 1 to n, where n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0025] Item 2. The use of an oligosaccharide formulation for treating, alleviating, preventing, and / or delaying said respiratory diseases, preferably infectious bronchitis (particularly infectious bronchitis caused by IBV) and / or their effects (i.e., effects on said respiratory diseases, effects caused by said respiratory diseases, or effects related to said respiratory diseases) in animals (preferably poultry, more preferably chickens) suffering from and / or at risk of said respiratory diseases; wherein said oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having different degrees of polymerization selected from 1 to n, where n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0026] Item 3. Use of an oligosaccharide formulation for improving the efficacy, effectiveness, and / or efficacy of a vaccine against respiratory diseases, preferably infectious bronchitis (particularly infectious bronchitis caused by IBV); wherein the oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having a different degree of polymerization selected from 1 to n, where n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of an oligosaccharide containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0027] Item 4. Use of an oligosaccharide formulation for reducing, preventing and / or delaying mortality in poultry populations challenged by respiratory diseases, preferably infectious bronchitis (particularly infectious bronchitis caused by IBV); wherein the oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having different degrees of polymerization selected from 1 to n, where n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0028] Item 5. Use of an oligosaccharide formulation for the treatment, mitigation, prevention and / or delay of the incidence of respiratory diseases, preferably infectious bronchitis (particularly infectious bronchitis caused by IBV); wherein the oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having different degrees of polymerization selected from 1 to n, where n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0029] Item 6. The use according to any one of the preceding items, wherein the oligosaccharide preparation is included in the nutritional composition at a concentration of greater than 40 ppm (e.g., greater than 40 ppm, 50 ppm, 60 ppm, 70 ppm, 80 ppm, 90 ppm, 100 ppm, 110 ppm, 120 ppm, 130 ppm, 140 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm, 1000 ppm, 1100 ppm, 1200 ppm, etc.).

[0030] Item 7. The use according to any one of the preceding items, wherein the n of the oligosaccharide formulation is at least 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, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100.

[0031] Item 8. The use according to any one of the preceding items, wherein at least one fraction of the oligosaccharide formulation comprises less than 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% of oligosaccharides containing dehydrated subunits in relative abundance; and / or wherein each fraction of the oligosaccharide formulation comprises more than 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, 50%, 60%, 70% or 80% of oligosaccharides containing dehydrated subunits in relative abundance.

[0032] Item 9. The use according to any one of the preceding items, wherein the weight-average molecular weight of said oligosaccharide formulation is about 300 to 5000 g / mol, for example about 2000 to 2800 g / mol, 2100 to 2700 g / mol, 2200 to 2600 g / mol, 2300 to 2500 g / mol, or 2320 to 2420 g / mol; 500 to 5000 g / mol, 700 to 5000 g / mol, 500 to 2000 g / mol, 700 to 2000 g / mol, 700 to 1500 g / mol, 300 to 1500 g / mol, 300 to 2000 g / mol, 400 to 1300 g / mol, 400 to 1200 g / mol, 400 to 1100 g / mol, 500 to 1300 g / mol, 500 to 12 ...300 g / mol, 500 to 1200 g / mol, 500 to 1200 g / mol, 500 to 1300 g / mol, 500 to 1200 g / mol, 500 to 1300 g / mol, 500 to 1200 g / mol, 500 to 1300 g / mol, 500 to g / mol, 500 to 1100 g / mol, 600 to 1300 g / mol, 600 to 1200 g / mol or 600 to 1100 g / mol; and / or the number average molecular weight of said oligosaccharide formulation is about 1000 to 2000 g / mol, 1100 to 1900 g / mol, 1200 to 1800 g / mol, 1300 to 1700 g / mol, 1400 to 1600 g / mol or 1450 to 1550 g / mol.

[0033] Item 10. The use according to any one of the preceding items, wherein the relative abundance of oligosaccharides in each of the n fractions of the oligosaccharide formulation decreases monotonically with its degree of polymerization.

[0034] Item 11. The use according to any one of the preceding items, wherein the relative abundance of oligosaccharides in at least 5, 10, 20, and 30 DP fractions of the oligosaccharide formulation decreases monotonically with their degree of polymerization.

[0035] Item 12. The use according to any one of the preceding items, wherein the oligosaccharide formulation comprises less than 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% of anhydrous oligosaccharides.

[0036] Item 13. The use according to any one of the preceding items, wherein each fraction of the oligosaccharide formulation comprises less than 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% of anhydrous oligosaccharides.

[0037] Item 14. The use according to any one of the preceding items, wherein at least one fraction of the oligosaccharide formulation comprises, in a relative abundance of more than 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 40%, 50%, 60%, 70% or 80% of oligosaccharides containing dehydrated subunits.

[0038] Item 15. The use according to any one of the preceding items, wherein the oligosaccharide formulation comprises, in a relative abundance of more than 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, or 25%, 30%, 40%, 50%, 60%, 70%, or 80% of anhydrous oligosaccharides.

[0039] Item 16. The use according to any one of the preceding items, wherein each fraction of the oligosaccharide formulation comprises, in a relative abundance of more than 20%, 21%, 22%, 23%, 24% or 25% of anhydrous oligosaccharides.

[0040] Item 17. The use according to any one of the preceding items, wherein more than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, or 30% of the oligosaccharide containing dehydrated subunits of the oligosaccharide preparation has only one dehydrated subunit.

[0041] Item 18. The use according to any one of the preceding items, wherein the oligosaccharide formulation has a DP1 fraction content of 1% to 40% in terms of relative abundance.

[0042] Item 19. The use according to any one of the preceding items, wherein the oligosaccharide formulation has a DP2 fraction content of 1% to 35% in terms of relative abundance.

[0043] Item 20. The use according to any one of the preceding items, wherein the oligosaccharide formulation has a DP3 fraction content of 1% to 30% in terms of relative abundance.

[0044] Item 21 The use according to any one of the preceding items, wherein the oligosaccharide formulation has a DP4 fraction content of 0.1% to 20% in terms of relative abundance.

[0045] Item 22. The use according to any one of the preceding items, wherein the oligosaccharide formulation has a DP5 fraction content of 0.1% to 15% in relative abundance.

[0046] Item 23. The use according to any one of the preceding items, wherein the ratio of the DP2 fraction to the DP1 fraction of the oligosaccharide formulation is 0.02 to 0.40 in terms of relative abundance.

[0047] Item 24. The use according to any one of the preceding items, wherein the ratio of the DP3 fraction to the DP2 fraction of the oligosaccharide formulation is 0.01 to 0.30 in terms of relative abundance.

[0048] Item 25. For any of the preceding items, the content of the aggregates of the DP1 and DP2 fractions in the oligosaccharide formulation is less than 50%, 30%, or 10% in terms of relative abundance.

[0049] Item 26. The use according to any one of the preceding items, wherein the oligosaccharide preparation comprises at least 103, 104, 105, 106 or 109 different oligosaccharide species.

[0050] Item 27. The use according to any one of the preceding items, wherein the two or more independent oligosaccharides of the oligosaccharide formulation comprise different dehydrated subunits.

[0051] Item 28. The use according to any one of the preceding items, wherein the oligosaccharide formulation comprises one or more dehydrated subunits, said one or more dehydrated subunits being products of reversible thermal dehydration of a monosaccharide.

[0052] Item 29. The use according to any one of the preceding items, wherein the oligosaccharide preparation comprises one or more of dehydrated glucose, dehydrated galactose, dehydrated mannose, dehydrated allose, dehydrated arabinose, anhydro-gulose, anhydro-indose, dehydrated tarose, dehydrated fructose, dehydrated ribose, dehydrated arabinose, dehydrated rhamnose, anhydro-lyxose, or dehydrated xylose subunits.

[0053] Item 30. The use according to any one of the preceding items, wherein the oligosaccharide preparation comprises one or more dehydrated glucose, dehydrated galactose, dehydrated mannose or defructose subunits.

[0054] Item 31. The use according to any one of the preceding items, wherein the oligosaccharide formulation comprises one or more 1,6-dehydro-β-D-furanose or 1,6-dehydro-β-D-pyranose subunits. In some embodiments, the oligosaccharide formulation comprises both 1,6-dehydro-β-D-furanose and 1,6-dehydro-β-D-pyranose dehydrated subunits.

[0055] Item 32. The use according to any one of the preceding items, wherein in the oligosaccharide formulation, the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose is about 10:1 to 1:10, 9:1 to 1:10, 8:1 to 1:10, 7:1 to 1:10, 6:1 to 1:10, 5:1 to 1:10, 4:1 to 1:10, 3:1 to 1:10, 2:1 to 1:10, 10:1 to 1:9, 10:1 to 1:8, 10:1 to 1:7, 10:1 to 1:6, 10:1 to 1:5, 10:1 to 1:4, 10:1 to 1:3, 10:1 to 1:2 or 1:1 to 3:1.

[0056] Item 33. The use according to any one of the preceding items, wherein in the oligosaccharide formulation, the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose is about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

[0057] Item 34. The use according to any one of the preceding items, wherein the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose in the oligosaccharide preparation is about 2:1.

[0058] Item 35. The use according to any one of the preceding items, wherein in each fraction of the oligosaccharide formulation, the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose is about 10:1 to 1:10, 9:1 to 1:10, 8:1 to 1:10, 7:1 to 1:10, 6:1 to 1:10, 5:1 to 1:10, 4:1 to 1:10, 3:1 to 1:10, 2:1 to 1:10, 10:1 to 1:9, 10:1 to 1:8, 10:1 to 1:7, 10:1 to 1:6, 10:1 to 1:5, 10:1 to 1:4, 10:1 to 1:3, 10:1 to 1:2 or 1:1 to 3:1.

[0059] Item 36. The use according to any one of the preceding items, wherein in each fraction of the oligosaccharide formulation, the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose is about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

[0060] Item 37. The use according to any one of the preceding items, wherein in each fraction of the oligosaccharide formulation, the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose is about 2:1.

[0061] Item 38. The use according to any one of the preceding items, wherein at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% of the dehydrated subunits in the oligosaccharide formulation are selected from the group consisting of 1,6-dehydrated-β-D-furanose and 1,6-dehydrated-β-D-pyranose.

[0062] Item 39. The use according to any one of the preceding items, wherein the weight-average molecular weight of said oligosaccharide preparation is about 300 to 5000 g / mol, 500 to 5000 g / mol, 700 to 5000 g / mol, 500 to 2000 g / mol, 700 to 2000 g / mol, 700 to 1500 g / mol, 300 to 1500 g / mol, 300 to 2000 g / mol, 400 to 1300 g / mol, 400 to 1200 g / mol, 400 to 1100 g / mol, 500 to 1300 g / mol, 500 to 1200 g / mol, 500 to 1100 g / mol, 600 to 1300 g / mol, 600 to 1200 g / mol, or 600 to 1100 g / mol.

[0063] Item 40. The use according to any one of the preceding items, wherein the number average molecular weight of said oligosaccharide preparation is about 300 to 5000 g / mol, 500 to 5000 g / mol, 700 to 5000 g / mol, 500 to 2000 g / mol, 700 to 2000 g / mol, 700 to 1500 g / mol, 300 to 1500 g / mol, 300 to 2000 g / mol, 400 to 1000 g / mol, 400 to 900 g / mol, 400 to 800 g / mol, 500 to 900 g / mol, or 500 to 800 g / mol.

[0064] Item 41. The use according to any one of the preceding items, wherein the distribution of the degree of polymerization is determined and / or detected by MALDI-MS, GC-MS, LC-MS, SEC, HPLC and / or combinations thereof (e.g., MALDI-MS and SEC).

[0065] Item 42. The use according to any one of the preceding items, wherein the degree of polymerization of the oligosaccharide formulation can be determined based on its molecular weight and molecular weight distribution.

[0066] Item 43. The use according to any one of the preceding items, wherein the oligosaccharide preparation is contained in the nutritional composition at a concentration of at least 50 g per ton of feed (e.g., at least 70 g, 100 g, 200 g, 300 g, 400 g, 500 g, 600 g, 700 g, 800 g, or 900 g per ton of feed); and / or wherein the oligosaccharide preparation is contained in the nutritional composition at a concentration of at least 50 ppm (e.g., at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, or 500 ppm); and / or wherein the oligosaccharide preparation is contained in the nutritional composition at a concentration of at least 50 ppm (e.g., at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, or 500 ppm).

[0067] Item 44. The use according to any one of the preceding items, wherein the oligosaccharide preparation is fed for at least one day, preferably at least 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, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 6 8, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 1 The nutritional composition is fed continuously (i.e., without interruption) for 93, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, or 245 days, most preferably continuously (i.e., without interruption).

[0068] Item 44. The use according to any one of the preceding items, wherein the average DP of the oligosaccharide preparation is at least 2, or at least 3, or at least 4.

[0069] Item 44. The use according to any one of the preceding items, wherein the average DP of the oligosaccharide preparation is less than 5.0.

[0070] Item 44. The use according to any one of the preceding items, wherein the average DP of the oligosaccharide preparation is greater than or equal to 2, for example at least 3, or at least 4; and / or wherein the average DP of the oligosaccharide preparation is less than 5.0.

[0071] Item 44. The use according to any one of the preceding items, wherein the average DP of the formulation is determined based on its number-average molecular weight distribution.

[0072] Item 45. An oligosaccharide preparation for use in methods of treating, alleviating, preventing and / or delaying respiratory diseases, preferably infectious bronchitis (particularly infectious bronchitis caused by IBV) and / or their effects (i.e., the effects of said respiratory disease, or the effects caused by said respiratory disease, or the effects associated with said respiratory disease); wherein said oligosaccharide preparation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having different degrees of polymerization selected from 1 to n, where n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0073] Item 46. An oligosaccharide formulation for use in treating, alleviating, preventing, and / or delaying respiratory diseases, preferably infectious bronchitis (particularly infectious bronchitis caused by IBV), and / or their effects (i.e., effects of the respiratory disease, effects caused by the respiratory disease, or effects related to the respiratory disease) in animals suffering from and / or at risk of suffering from said respiratory diseases; wherein said oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having different degrees of polymerization selected from 1 to n, where n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

[0074] Item 47. An oligosaccharide preparation used according to any one of items 45 or 46, wherein the oligosaccharide preparation is administered in addition to vaccination against the respiratory disease.

[0075] Item 48. An oligosaccharide formulation used according to any one of items 45-47, wherein the oligosaccharide formulation is included in the nutritional composition at a concentration of greater than 40 ppm (e.g., greater than 40 ppm, 50 ppm, 60 ppm, 70 ppm, 80 ppm, 90 ppm, 100 ppm, 110 ppm, 120 ppm, 130 ppm, 140 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm, 1000 ppm, 1100 ppm, 1200 ppm, etc.).

[0076] Item 49. An oligosaccharide formulation used according to any one of items 45-48, wherein the n of the oligosaccharide formulation is at least 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, 4 6, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100.

[0077] Item 50. An oligosaccharide formulation for use according to any one of items 45-49, wherein at least one fraction of the oligosaccharide formulation comprises less than 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% of oligosaccharides containing dehydrated subunits in relative abundance; and / or wherein each fraction of the oligosaccharide formulation comprises more than 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, 50%, 60%, 70% or 80% of oligosaccharides containing dehydrated subunits in relative abundance.

[0078] Item 51. An oligosaccharide formulation for use according to any one of items 45-50, wherein the weight-average molecular weight of said oligosaccharide formulation is about 300 to 5000 g / mol, for example about 2000 to 2800 g / mol, 2100 to 2700 g / mol, 2200 to 2600 g / mol, 2300 to 2500 g / mol or 2320 to 2420 g / mol; 500 to 5000 g / mol, 700 to 5000 g / mol, 500 to 2000 g / mol, 700 to 2000 g / mol, 700 to 1500 g / mol, 300 to 1500 g / mol, 300 to 2000 g / mol, 400 to 1300 g / mol, 400 to 1200 g / mol, 400 to 1100 g / mol, 500 to 1300 g / mol, 400 to 1200 g / mol, 400 to 1100 g / mol, 500 to 1300 g / mol. g / mol, 500 to 1200 g / mol, 500 to 1100 g / mol, 600 to 1300 g / mol, 600 to 1200 g / mol or 600 to 1100 g / mol; and / or the number average molecular weight of said oligosaccharide formulation is about 1000 to 2000 g / mol, 1100 to 1900 g / mol, 1200 to 1800 g / mol, 1300 to 1700 g / mol, 1400 to 1600 g / mol or 1450 to 1550 g / mol.

[0079] Item 52. An oligosaccharide formulation used according to any one of items 45-51, wherein the relative abundance of oligosaccharides in each of the n fractions of the oligosaccharide formulation decreases monotonically with its degree of polymerization.

[0080] Item 53. An oligosaccharide formulation used according to any one of items 45-52, wherein the relative abundance of oligosaccharides in at least 5, 10, 20, and 30 DP fractions of the oligosaccharide formulation decreases monotonically with their degree of polymerization.

[0081] Item 54. An oligosaccharide formulation for use according to any one of items 45-53, wherein the oligosaccharide formulation comprises, in a relative abundance of less than 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, or 2% of anhydrous oligosaccharides.

[0082] Item 55. An oligosaccharide formulation for use according to any one of items 45-54, wherein each fraction of the oligosaccharide formulation comprises less than 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% of anhydrous oligosaccharides.

[0083] Item 56. An oligosaccharide formulation for use according to any one of items 45-55, wherein at least one fraction of the oligosaccharide formulation comprises, in a relative abundance greater than 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 40%, 50%, 60%, 70%, or 80% of anhydrous oligosaccharides.

[0084] Item 57. An oligosaccharide formulation for use according to any one of items 45-56, wherein the oligosaccharide formulation comprises, in a relative abundance greater than 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, or 25%, 30%, 40%, 50%, 60%, 70%, or 80% of anhydrous oligosaccharides.

[0085] Item 58. An oligosaccharide formulation for use according to any one of items 45-57, wherein each fraction of the oligosaccharide formulation comprises, in a relative abundance of more than 20%, 21%, 22%, 23%, 24% or 25% of anhydrous oligosaccharides.

[0086] Item 59. An oligosaccharide formulation used according to any one of items 45-58, wherein more than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, or 30% of the oligosaccharide containing dehydrated subunits of the oligosaccharide formulation has only one dehydrated subunit.

[0087] Item 60. An oligosaccharide formulation used according to any one of items 45-59, wherein the oligosaccharide formulation has a DP1 fraction content of 1% to 40% in terms of relative abundance.

[0088] Item 61. An oligosaccharide formulation used according to any one of items 45-60, wherein the oligosaccharide formulation has a DP2 fraction content of 1% to 35% in terms of relative abundance.

[0089] Item 62. An oligosaccharide formulation used according to any one of items 45-61, wherein the oligosaccharide formulation has a DP3 fraction content of 1% to 30% in terms of relative abundance.

[0090] Item 63. An oligosaccharide formulation for use according to any one of items 45-62, wherein the oligosaccharide formulation has a DP4 fraction content of 0.1% to 20% in terms of relative abundance.

[0091] Item 64. An oligosaccharide formulation used according to any one of items 45-63, wherein the oligosaccharide formulation has a DP5 fraction content of 0.1% to 15% in relative abundance.

[0092] Item 65. An oligosaccharide formulation used according to any one of items 45-64, wherein the ratio of the DP2 fraction to the DP1 fraction of the oligosaccharide formulation is from 0.02 to 0.40 in terms of relative abundance.

[0093] Item 66. An oligosaccharide formulation used according to any one of items 45-65, wherein the ratio of the DP3 fraction to the DP2 fraction of the oligosaccharide formulation is from 0.01 to 0.30 in terms of relative abundance.

[0094] Item 67. An oligosaccharide formulation used according to any one of items 45-66, wherein the content of aggregates of DP1 and DP2 fractions in the oligosaccharide formulation is less than 50%, 30% or 10% in relative abundance.

[0095] Item 68. An oligosaccharide formulation used according to any one of items 45-67, wherein the oligosaccharide formulation comprises at least 103, 104, 105, 106 or 109 different oligosaccharide species.

[0096] Item 69. An oligosaccharide formulation used according to any one of items 45-68, wherein the two or more independent oligosaccharides of the oligosaccharide formulation comprise different dehydrated subunits.

[0097] Item 70. An oligosaccharide formulation used according to any one of items 45-69, wherein the oligosaccharide formulation comprises one or more dehydrated subunits, the one or more dehydrated subunits being products of reversible thermal dehydration of a monosaccharide.

[0098] Item 71. An oligosaccharide preparation used according to any one of Items 45-70, wherein the oligosaccharide preparation comprises one or more of dehydrated glucose, dehydrated galactose, dehydrated mannose, dehydrated allose, dehydrated arabinose, anhydro-gulose, anhydro-indose, dehydrated tarose, dehydrated fructose, dehydrated ribose, dehydrated arabinose, dehydrated rhamnose, anhydro-lyxose, or dehydrated xylose subunits.

[0099] Item 72. An oligosaccharide preparation used according to any one of Items 45-71, wherein the oligosaccharide preparation comprises one or more dehydrated glucose, dehydrated galactose, dehydrated mannose, or defructose subunits.

[0100] Item 73. An oligosaccharide formulation used according to any one of items 45-72, wherein the oligosaccharide formulation comprises one or more 1,6-dehydro-β-D-furanose or 1,6-dehydro-β-D-pyranose subunits. In some embodiments, the oligosaccharide formulation comprises both 1,6-dehydro-β-D-furanose and 1,6-dehydro-β-D-pyranose dehydrated subunits.

[0101] Item 74. An oligosaccharide formulation for use according to any one of items 45-73, wherein the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose in the oligosaccharide formulation is about 10:1 to 1:10, 9:1 to 1:10, 8:1 to 1:10, 7:1 to 1:10, 6:1 to 1:10, 5:1 to 1:10, 4:1 to 1:10, 3:1 to 1:10, 2:1 to 1:10, 10:1 to 1:9, 10:1 to 1:8, 10:1 to 1:7, 10:1 to 1:6, 10:1 to 1:5, 10:1 to 1:4, 10:1 to 1:3, 10:1 to 1:2 or 1:1 to 3:1.

[0102] Item 75. An oligosaccharide formulation for use according to any one of items 45-74, wherein the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose in the oligosaccharide formulation is about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

[0103] Item 76. An oligosaccharide formulation used according to any one of items 45-75, wherein the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose in the oligosaccharide formulation is about 2:1.

[0104] Item 77. An oligosaccharide formulation for use according to any one of items 45-76, wherein in each fraction of the oligosaccharide formulation, the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose is about 10:1 to 1:10, 9:1 to 1:10, 8:1 to 1:10, 7:1 to 1:10, 6:1 to 1:10, 5:1 to 1:10, 4:1 to 1:10, 3:1 to 1:10, 2:1 to 1:10, 10:1 to 1:9, 10:1 to 1:8, 10:1 to 1:7, 10:1 to 1:6, 10:1 to 1:5, 10:1 to 1:4, 10:1 to 1:3, 10:1 to 1:2 or 1:1 to 3:1.

[0105] Item 78. An oligosaccharide formulation for use according to any one of items 45-77, wherein in each fraction of the oligosaccharide formulation, the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose is about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

[0106] Item 79. An oligosaccharide formulation used according to any one of items 45-78, wherein in each fraction of the oligosaccharide formulation, the ratio of 1,6-dehydro-β-D-furanose glucose to 1,6-dehydro-β-D-pyranose glucose is about 2:1.

[0107] Item 80. An oligosaccharide formulation used according to any one of items 45-79, wherein at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% of the dehydrated subunits in the oligosaccharide formulation are selected from the group consisting of 1,6-dehydrated-β-D-furanose and 1,6-dehydrated-β-D-pyranose.

[0108] Item 81. An oligosaccharide formulation used according to any one of items 45-80, wherein the weight-average molecular weight of the oligosaccharide formulation is about 300 to 5000 g / mol, 500 to 5000 g / mol, 700 to 5000 g / mol, 500 to 2000 g / mol, 700 to 2000 g / mol, 700 to 1500 g / mol, 300 to 1500 g / mol, 300 to 2000 g / mol, 400 to 1300 g / mol, 400 to 1200 g / mol, 400 to 1100 g / mol, 500 to 1300 g / mol, 500 to 1200 g / mol, 500 to 1100 g / mol, 600 to 1300 g / mol, 600 to 1200 g / mol, or 600 to 1100 g / mol.

[0109] Item 82. An oligosaccharide formulation used according to any one of items 45-81, wherein the number average molecular weight of said oligosaccharide formulation is about 300 to 5000 g / mol, 500 to 5000 g / mol, 700 to 5000 g / mol, 500 to 2000 g / mol, 700 to 2000 g / mol, 700 to 1500 g / mol, 300 to 1500 g / mol, 300 to 2000 g / mol, 400 to 1000 g / mol, 400 to 900 g / mol, 400 to 800 g / mol, 500 to 900 g / mol, or 500 to 800 g / mol.

[0110] Item 83. The oligosaccharide formulation used according to any one of items 45-82, wherein the distribution of the degree of polymerization is determined and / or detected by MALDI-MS, GC-MS, LC-MS, SEC, HPLC and / or combinations thereof (e.g., MALDI-MS and SEC).

[0111] Item 84. An oligosaccharide formulation used according to any one of items 45-83, wherein the degree of polymerization of the oligosaccharide formulation can be determined based on its molecular weight and molecular weight distribution.

[0112] Item 85. An oligosaccharide preparation used according to any one of Items 45-84, wherein the oligosaccharide preparation is contained in the nutritional composition at a concentration of at least 50 g per tonne of feed (e.g., at least 70 g, 100 g, 200 g, 300 g, 400 g, 500 g, 600 g, 700 g, 800 g, or 900 g per tonne of feed); and / or wherein the oligosaccharide preparation is contained in the nutritional composition at a concentration of at least 50 ppm (e.g., at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, or 500 ppm); and / or wherein the oligosaccharide preparation is contained in the nutritional composition at a concentration of at least 50 ppm (e.g., at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, or 500 ppm).

[0113] Item 86. An oligosaccharide preparation used according to any one of items 45-85, wherein the oligosaccharide preparation is fed for at least one day, preferably at least 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, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 6 6, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192 The nutritional composition is fed continuously (i.e., without interruption) for 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, or 245 days, most preferably continuously (i.e., without interruption).

[0114] Item 87. An oligosaccharide formulation used according to any one of items 45-86, wherein the average DP of the oligosaccharide formulation is at least 2, or at least 3, or at least 4.

[0115] Item 88. An oligosaccharide formulation used according to any one of items 45-87, wherein the average DP of the oligosaccharide formulation is less than 5.0.

[0116] Item 89. An oligosaccharide formulation used according to any one of items 45-88, wherein the average DP of the oligosaccharide formulation is greater than or equal to 2, for example at least 3, or at least 4; and / or wherein the average DP of the oligosaccharide formulation is less than 5.0.

[0117] Item 90. An oligosaccharide formulation used according to any one of items 45-89, wherein the average DP of the formulation is determined based on its number-average molecular weight distribution.

[0118] It is also believed that the oligosaccharide formulations of the present invention can be provided in the form of a powder formulation comprising at least 20% (w / w) of the oligosaccharide formulations mentioned herein; at least 25% (wt / wt) of a silica-based adsorbent (e.g., diatomaceous earth, amorphous precipitated silica) with an average particle size D less than or equal to 3000 μm (e.g., 100-500 μm, 200-500 μm, 200-300 µm, etc.); and optionally 0-25% (wt / wt) of water and / or auxiliary substances; wherein the percentage is based on the total weight of the powder formulation. For example, such a powder formulation may comprise 30-70% (wt / wt) of the oligosaccharide formulations mentioned herein; 30-70% (wt / wt) of a silica-based adsorbent (e.g., having an average particle size of at least 50 μm); and 0-21% (wt / wt) of water; wherein the percentage is based on the total weight of the powder formulation. In some embodiments, the oligosaccharide formulation is formulated as described in any one of Examples 22 to 26 and Example 33 of WO 2020 / 097458.

[0119] Methods for manufacturing oligosaccharide formulations according to the invention are described in WO 2020 / 097458, WO 2016 / 007778, and particularly in the examples described therein, especially in any one of Examples 1-7, 16-18 of WO 2020 / 097458 A1; in the methods described in paragraph

[317] of WO 2016 / 007778 A1 and / or in any one of Examples 73-77, 80-89, 97-99, 101-110; preferably, the oligosaccharide formulation according to the invention is manufactured as described in Example 16 of WO 2020 / 097458A1. In particular, the oligosaccharide formulation according to the invention is characterized by the step of heating an aqueous composition comprising one or more feed sugars and a catalyst to a temperature and time sufficient to induce polymerization. In some embodiments, the feed sugar is glucose, mannose, galactose, lactose, fructose, sucrose, xylose, arabinose, glucosamine, N-acetylglucosamine, N-acetylglucosamine, glucuronic acid, galacturonic acid, fucose, rhamnose, chinovose, or any combination thereof; glucose and mannose, glucose and galactose, glucose and lactose, glucose and fructose, glucose and arabinose, glucose and sucrose, glucose and xylose, glucose and galactose and arabinose, etc.In some embodiments, the catalyst is any one of the following: (+)-camphor-10-sulfonic acid; 2-pyridinesulfonic acid; 3-pyridinesulfonic acid; 8-hydroxy-5-quinolinesulfonic acid hydrate; α-hydroxy-2-pyridinemethanesulfonic acid; (β)-camphor-10-sulfonic acid; butylphosphonic acid; diphenylphosphonic acid; hexylphosphonic acid; methylphosphonic acid; phenylphosphonic acid; phenylphosphonic acid; tert-butylphosphonic acid; SS)-VAPOL hydrogen phosphate; 6-quinolinesulfonic acid; 3-(1-pyridyl)-1-propanesulfonate. ; 2-(2-pyridyl)ethanesulfonic acid; 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine-p,p'-disulfonic acid monosodium hydrate; 1,1'-binaphthyl-2,2'-diyl-hydrogen phosphate; bis(4-methoxyphenyl)phosphinoic acid; phenyl(3,5-dimethyl)phosphinoic acid; L-sulfoalanine monohydrate; poly(styrenesulfonic acid-copolymer-divinylbenzene); lysine; ethanedisulfonic acid; ethanesulfonic acid; hydroxyethanesulfonic acid (isethionic) (acid); homocysteine; HEPBS (N-(2-hydroxyethyl)piperazine-N'-(4-butyric acid)); HEPES (4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid); 2-hydroxy-3-morpholinopropanesulfonic acid; 2-(N-morpholino)ethanesulfonic acid; methanesulfonic acid; nicotinamide methanesulfonate; naphthalene-1-sulfonic acid; naphthalene-2-sulfonic acid; perfluorobutyric acid; 6-sulfoquinolate; trifluoromethanesulfonic acid; 2-aminoethanesulfonic acid; benzoic acid; chloroacetic acid; trifluoroacetic acid; hexanoic acid; heptanoic acid; octanoic acid; nonanoic acid; lauric acid; palmitic acid; stearic acid; arachidic acid; aspartic acid; glutamic acid; serine; threonine; glutamine; cysteine; glycine; proline; alanine; valine; isoleucine; leucine; methionine; phenylalanine; tyrosine; or tryptophan. In some embodiments, the feed sugar is glucose and the catalyst is (+)-camphor-10-sulfonic acid, (β)-camphor-10-sulfonic acid, butylphosphonic acid, HEPBS, naphthalene-1-sulfonic acid, naphthalene-2-sulfonic acid, or palmitic acid. In some embodiments, the feed sugar is glucose and mannose, and the catalyst is ethanedisulfonic acid, ethanesulfonic acid, trifluoromethanesulfonic acid, or lauric acid. In some embodiments, the feed sugar is galactose, and the catalyst is 2-(N-morpholino)ethanesulfonic acid, methanesulfonic acid, or nicotinamide methanesulfonate.

[0120] For clarity only, the term "oligosaccharide formulation" may refer to a formulation containing one or more oligosaccharides.

[0121] As used herein, "oligosaccharide" or "oligomery" can refer to a monosaccharide unit or a compound containing two or more monosaccharide subunits linked by glycosidic bonds to form an oligosaccharide. "Oligosaccharide" can also refer to anhydrous monosaccharide units or a compound containing two or more monosaccharide subunits, wherein at least one monosaccharide unit is replaced by anhydrous subunits. "Oligosaccharide" may optionally be functionalized. As used herein, the term "oligosaccharide" encompasses all types of oligosaccharides in which each monosaccharide subunit is independently and optionally functionalized and / or replaced by its corresponding anhydrous monosaccharide subunit.

[0122] "Dehydrated subunit" can be a monosaccharide or a monosaccharide subunit or a reversible thermal dehydration product of a caramelized sugar. For example, "dehydrated subunit" can be a dehydrated monosaccharide, such as dehydrated glucose. As another example, "dehydrated subunit" can be linked to one or more conventional monosaccharide subunits and / or to one or more dehydrated monosaccharide subunits via a glycosidic bond.

[0123] Oligosaccharides are characterized by containing two or more monosaccharide subunits linked by glycosidic bonds. In this regard, the term "glucose oligosaccharide" can refer to a single glucose molecule or a compound containing two or more glucose monosaccharide subunits linked by glycosidic bonds. "Glucose oligosaccharide" can also refer to dehydrated glucose or a compound containing two or more glucose monosaccharide subunits linked by glycosidic bonds, wherein at least one monosaccharide subunit is replaced by a dehydrated glucose subunit. Similarly, "galactose oligosaccharide" can refer to galactose or a compound containing two or more galactose monosaccharide subunits linked by glycosidic bonds. "Galactose oligosaccharide" can also refer to dehydrated galactose or a compound containing two or more galactose monosaccharide subunits linked by glycosidic bonds, wherein at least one monosaccharide subunit is replaced by a dehydrated galactose subunit. Similarly, glucose-galactose-oligosaccharide can be a compound containing one or more glucose monosaccharide subunits and one or more galactose monosaccharide subunits linked by glycosidic bonds. Glucose-galactose oligosaccharides can also refer to compounds in which at least one monosaccharide subunit is replaced by its corresponding dehydrated monosaccharide subunit. Glucose-galactose-xylose-oligosaccharides can refer to compounds produced by the condensation reaction of glucose, galactose, and xylose. Oligosaccharide preparations containing glucose-galactose-xylose-oligosaccharides can include glucose-galactose-oligosaccharides, glucose-xylose-oligosaccharides, galactose-xylose-oligosaccharides, glucose-galactose-xylose-oligosaccharides, and compounds containing one or more glucose monosaccharide subunits, one or more xylose monosaccharide subunits, and one or more galactose monosaccharide subunits and their dehydrated monosaccharide subunits linked by glycosidic bonds.

[0124] As used herein, unless otherwise stated, the terms "monosaccharide unit" and "monosaccharide subunit" are used interchangeably. "Monosaccharide subunit" can refer to the monosaccharide monomer in an oligosaccharide. For an oligosaccharide with a degree of polymerization (DP) of 1, the oligosaccharide can be referred to as a monosaccharide subunit or simply a monosaccharide. For oligosaccharides with a degree of polymerization greater than 1, the monosaccharide subunits are linked via glycosidic bonds.

[0125] As used herein, the term "conventional monosaccharide" may refer to a monosaccharide that does not contain anhydrous subunits. The term "conventional disaccharide" may refer to a disaccharide that does not contain anhydrous subunits. Therefore, the term "conventional subunit" may refer to a subunit that is not anhydrous subunits.

[0126] As used herein, the terms “relative abundance” or “abundance” may refer to the abundance of a substance in terms of its prevalence or rarity. For example, a DP1 fraction (i.e., a fraction with DP of 1) containing 10% of anhydrous subunit-containing oligosaccharides in terms of relative abundance may refer to a variety of DP1 oligosaccharides, wherein 10% by quantity of the DP1 oligosaccharides are anhydrous monosaccharides.

[0127] Distribution of degree of aggregation (DP) The degree of polymerization (DOP) distribution of oligosaccharide formulations can be determined by any suitable analytical method and instrument, including but not limited to end-group methods, osmometry (osmoregulation), ultracentrifugation, viscosity measurement, light scattering, size exclusion chromatography (SEC), SEC-MALLS, field flow fractionation (FFF), asymmetric flow field flow fractionation (A4F), high-performance liquid chromatography (HPLC), and mass spectrometry (MS). For example, the DOP distribution can be determined and / or detected by mass spectrometry (e.g., MALDI-MS, LC-MS, or GC-MS). As another example, the DOP distribution can be determined and / or detected by SEC (e.g., gel permeation chromatography (GPC)). As yet another example, the DOP distribution can be determined and / or detected by HPLC, FFF, or A4F. In yet another example, the DOP of oligosaccharide formulations can be determined based on their molecular weight and molecular weight distribution (see WO 2020 / 097458 for a more detailed description).

[0128] Dehydration subunit levelThe oligosaccharide formulation may comprise n distinct fractions, each fraction having a different DP (distribution density), and where n is any integer, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc. In some embodiments, each of the n oligosaccharide fractions of the oligosaccharide formulation described herein independently comprises a level of dehydrated subunits. For example, in some embodiments, fraction DP1 comprises 10% of a dehydrated subunit-containing oligosaccharides at a relative abundance, and fraction DP2 comprises 15% of a dehydrated subunit-containing oligosaccharides at a relative abundance. As another example, in some embodiments, fractions DP1, DP2, and DP3 each comprise 5%, 10%, and 2% of a dehydrated subunit-containing oligosaccharides at relative abundances, respectively. In other embodiments, two or more fractions of the oligosaccharide formulation may comprise similar levels of a dehydrated subunit-containing oligosaccharides. For example, in some embodiments, fractions DP1 and DP3 each comprise approximately 5% of a dehydrated subunit-containing oligosaccharides at a relative abundance.

[0129] The level of the dehydration subunit can be determined by any suitable analytical method, such as nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry, HPLC, FFF, A4F, or any combination thereof. In some embodiments, the level of the dehydration subunit is determined (at least partially) by mass spectrometry (e.g., MALDI-MS). In some embodiments, the level of the dehydration subunit may be determined (at least partially) by NMR. In some embodiments, the level of the dehydration subunit may be determined (at least partially) by HPLC. For example, in some embodiments, the level of the dehydration subunit can be determined by MALDI-MS, as described in more detail in WO 2020 / 097458.

[0130] glycosidic bond In some embodiments, the oligosaccharide formulations described herein comprise multiple glycosidic bonds. The type and distribution of glycosidic bonds may depend on the source and manufacturing method of the oligosaccharide formulation. In some embodiments, the type and distribution of various glycosidic bonds can be determined and / or detected by any suitable method known in the art (e.g., NMR). For example, in some embodiments, glycosidic bonds are determined and / or detected by proton NMR, carbon NMR, 2D NMR (e.g., 2D JRES, HSQC, HMBC, DOSY, COSY, ECOSY, TOCSY, NOESY, or ROESY) or any combination thereof. In some embodiments, glycosidic bonds are determined and / or detected (at least partially) by proton NMR. In some embodiments, glycosidic bonds are determined and / or detected (at least partially) by carbon NMR. In some embodiments, glycosidic bonds are determined and / or detected (at least partially) by 2D HSQC NMR.

[0131] In some embodiments, the oligosaccharide formulation may comprise one or more α-(1,2) glycosidic bonds, α-(1,3) glycosidic bonds, α-(1,4) glycosidic bonds, α-(1,6) glycosidic bonds, β-(1,2) glycosidic bonds, β-(1,3) glycosidic bonds, β-(1,4) glycosidic bonds, β-(1,6) glycosidic bonds, α(1,1)α glycosidic bonds, α(1,1)β glycosidic bonds, β(1,1)β glycosidic bonds, or any combination thereof.

[0132] In some embodiments, the glycosidic bond type distribution of the oligosaccharide formulation is approximately 0 mol% to 60 mol%, 5 mol% to 55 mol%, 5 mol% to 50 mol%, 5 mol% to 45 mol%, 5 mol% to 40 mol%, 5 mol% to 35 mol%, 5 mol% to 30 mol%, 5 mol% to 25 mol%, 10 mol% to 60 mol%, 10 mol% to 55 mol%, 10 mol% to 50 mol%, 10 mol% to 45 mol%, 10 mol% to 40 mol%, 10 mol% to 35 mol%, 15 mol% to 60 mol%, 15 mol% to 55 mol%, 15 mol% to 50 mol%, 15 mol% to 45 mol%, 15 mol% to 40 mol%, 15 mol% to 35 mol%, 20 mol% to 60 mol%, 20 mol% to 55 mol%. α-(1,6) glycosidic bonds of mol%, 20 mol% to 50 mol%, 20 mol% to 45 mol%, 20 mol% to 40 mol%, 20 mol% to 35 mol%, 25 mol% to 60 mol%, 25 mol% to 55 mol%, 25 mol% to 50 mol%, 25 mol% to 45 mol%, 25 mol% to 40 mol%, or 25 mol% to 35 mol%.

[0133] molecular weightThe molecular weight and molecular weight distribution of the oligosaccharide formulation can be determined by any suitable analytical method and instrument, such as end-group method, osmotic pressure (osmoregulation), ultracentrifugation, viscosity measurement, light scattering, SEC, SEC-MALLS, FFF, A4F, HPLC, and mass spectrometry. In some embodiments, the molecular weight and molecular weight distribution are determined by mass spectrometry (e.g., MALDI-MS, LC-MS, or GC-MS). In some embodiments, the molecular weight and molecular weight distribution are determined by size exclusion chromatography (SEC) (e.g., gel permeation chromatography (GPC)). In other embodiments, the molecular weight and molecular weight distribution are determined by HPLC. In some embodiments, the molecular weight and molecular weight distribution are determined by MALDI-MS.

[0134] In some embodiments, the weight-average molecular weight of the oligosaccharide formulation is about 100 to 10000 g / mol, 200 to 8000 g / mol, 300 to 5000 g / mol, 500 to 5000 g / mol, 700 to 5000 g / mol, 900 to 5000 g / mol, 1100 to 5000 g / mol, 1300 to 5000 g / mol, 1500 to 5000 g / mol, 1700 to 5000 g / mol, 300 to 4500 g / mol, 500 to 4500 g / mol, 700 to 4500 g / mol, 900 to 4500 g / mol, 1100 to 4500 g / mol, 1300 to 4500 g / mol, 1500 to 4500 g / mol, 1700 to 4500 g / mol. g / mol, 1900 to 4500 g / mol, 300 to 4000 g / mol, 500 to 4000 g / mol, 700 to 4000 g / mol, 900 to 4000 g / mol, 1100 to 4000 g / mol, 1300 to 4000 g / mol, 1500 to 4000 g / mol, 1700 to 4000 g / mol, 1900 to 4000 g / mol, 300 to 3000 g / mol, 500 to 3000 g / mol, 700 to 3000 g / mol, 900 to 3000 g / mol, 1100 to 3000 g / mol, 1300 to 3000 g / mol, 1500 to 3000 g / mol, 1700 to 3000 g / mol, 1900 to 3000 g / mol g / mol, 2100 to 3000 g / mol, 300 to 2500 g / mol, 500 to 2500 g / mol, 700 to 2500 g / mol, 900 to 2500 g / mol, 1100 to 2500 g / mol, 1300 to 2500 g / mol, 1500 to 2500 g / mol, 1700 to 2500 g / mol, 1900 to 2500 g / mol, 2100 to 2500 g / mol, 300 to 1500 g / mol, 500 to 1500 g / mol, 700 to 1500 g / mol, 900 to 1500 g / mol, 1100 to 1500 g / mol, 1300 to 1500 g / mol, 2000 to 2800 g / mol, 2100 to 2700 g / mol g / mol, 2200 to 2600 g / mol, 2300 to 2500 g / mol, or 2320 to 2420 g / mol.In some embodiments, the weight-average molecular weight of the oligosaccharide formulation is about 2000 to 2800 g / mol, 2100 to 2700 g / mol, 2200 to 2600 g / mol, 2300 to 2500 g / mol, or 2320 to 2420 g / mol.

[0135] Types of oligosaccharides In some embodiments, the types of oligosaccharides present in the oligosaccharide formulations mentioned herein may depend on the types of one or more feed sugars. For example, in some embodiments, when the feed sugar includes glucose, the oligosaccharide formulation comprises glucose-oligosaccharides. For example, in some embodiments, when the feed sugar includes galactose, the oligosaccharide formulation comprises galactose-oligosaccharides. For another example, in some embodiments, when the feed sugar includes both galactose and glucose, the oligosaccharide formulation comprises glucose-galactose-oligosaccharides.

[0136] In some embodiments, the oligosaccharide formulation comprises monosaccharide subunits of one or more species. In some embodiments, the oligosaccharide formulation may comprise oligosaccharides having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more different species of monosaccharide subunits.

[0137] Methods for manufacturing oligosaccharide preparationsThe method for manufacturing the oligosaccharide formulation according to the present invention is described in detail in WO 2020 / 097458, comprising heating an aqueous composition comprising one or more feed sugars and a catalyst to a temperature and time sufficient to induce polymerization, wherein the catalyst is selected from the group consisting of: (+)-camphor-10-sulfonic acid; 2-pyridinesulfonic acid; 3-pyridinesulfonic acid; 8-hydroxy-5-quinolinesulfonic acid hydrate; α-hydroxy-2-pyridinemethanesulfonic acid; (β)-camphor-10-sulfonic acid; butylphosphonic acid; diphenylphosphonic acid; hexylphosphonic acid; methylphosphonic acid. Acids; phenylphosphino acid; phenylphosphonic acid; tert-butylphosphonic acid; SS)-VAPOL hydrogen phosphate; 6-quinoline sulfonic acid, 3-(1-pyridyl)-1-propanesulfonate; 2-(2-pyridyl)ethanesulfonic acid; 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine-p,p'-disulfonic acid monosodium hydrate; 1,1'-binaphthyl-2,2'-diyl-hydrogen phosphate; bis(4-methoxyphenyl)phosphino acid; phenyl(3,5) -Xylyl)phosphonic acid; L-sulfoalanine monohydrate; poly(styrene sulfonic acid-co-divinylbenzene); lysine; ethanedisulfonic acid; ethanesulfonic acid; hydroxyethanesulfonic acid; homocysteine; HEPBS (N-(2-hydroxyethyl)piperazine-N'-(4-butyric acid)); HEPES (4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid); 2-hydroxy-3-morpholinopropanesulfonic acid; 2-(N-morpholino)ethanesulfonic acid; methanesulfonic acid; formylhydrazine Naphthalene-1-sulfonic acid; Naphthalene-2-sulfonic acid; Perfluorobutyric acid; 6-sulfoquinolate; Trifluoromethanesulfonic acid; 2-aminoethanesulfonic acid; Benzoic acid; Chloroacetic acid; Trifluoroacetic acid; Hexanoic acid; Heptanoic acid; Octanoic acid; Nonanoic acid; Lauric acid; Palmitic acid; Stearic acid; Arachidic acid; Aspartic acid; Glutamic acid; Serine; Threonine; Glutamine; Cysteine; Glycine; Proline; Alanine; Valine; Isoleucine; Leucine; Methionine; Phenylalanine; Tyrosine; Tryptophan.

[0138] In some embodiments, the polymerization of feed sugars is achieved through stepwise growth polymerization. In some embodiments, the polymerization of feed sugars is achieved through condensation polymerization.

[0139] Feed sugar The one or more feed sugars used in the method of manufacturing the oligosaccharide formulation described herein may comprise one or more types of sugars. In some embodiments, the one or more feed sugars include monosaccharides, disaccharides, trisaccharides, tetrasaccharides, or any mixture thereof.

[0140] In some embodiments, one or more feed sugars comprise glucose. In some embodiments, one or more feed sugars comprise glucose and galactose. In some embodiments, one or more feed sugars comprise glucose, xylose, and galactose. In some embodiments, one or more feed sugars comprise glucose and mannose. In some embodiments, one or more feed sugars comprise glucose and fructose. In some embodiments, one or more feed sugars comprise glucose, fructose, and galactose. In some embodiments, one or more feed sugars comprise glucose, galactose, and mannose.

[0141] When used herein, unless the context clearly indicates otherwise, the singular forms “a,” “an,” and “the,” as well as cases where no quantifier is used, include the plural referent. Thus, for example, reference to “a reagent” or “reagent” includes multiple such reagents, and reference to “oligosaccharide” includes reference to one or more oligosaccharides (or multiple oligosaccharides) and their equivalents known to those skilled in the art, and so on.

[0142] When ranges are used herein to refer to physical properties (e.g., molecular weight) or chemical properties (e.g., chemical formula), it is intended to include all combinations and sub-combinations of ranges, as well as specific embodiments thereof. When referring to a number or numerical range, the term “about” means that the number or numerical range mentioned is an approximation within experimental variability (or within statistical experimental error), and therefore in some cases, the number or numerical range will vary between 1% and 15% of said number or numerical range.

[0143] The invention is further described below by way of non-limiting embodiments.

[0144] Example The present invention disclosed herein is not limited to the specific embodiments, drawings, methodologies, examples, and solutions described herein, but is defined solely by the claims.

[0145] Example 1 For the broiler feeding trial, 32,600 male Ross broilers aged 308 days were fed either a control diet or an experimental diet. 10,800 broilers received the control diet, and 21,800 received the experimental diet. The control diet was the basal diet for broilers (corn and soybean meal). In addition to the basal diet, the experimental diet contained 0.9 kg / MT of an oligosaccharide formulation as described herein. Specifically, the oligosaccharide formulation contained more than two DP fractions and had an average DP of less than 5, as determined based on its number-average molecular weight distribution. Each DP fraction of the oligosaccharide formulation contained approximately 0.5%–15% of dehydrated subunit-containing oligosaccharides at relative abundance determined by mass spectrometry. Feeding was conducted according to common recommendations for this breed; the feed contained no antibiotic growth promoters or ionophores.

[0146] All birds were vaccinated against Newcastle disease, infectious bronchitis, NB, and IBQX. Feed and water were provided without restriction. Broiler (d0-10d) birds were fed crushed feed, grower (d11-d22) birds were fed pelleted feed, and finisher (d23-d35) birds were fed pelleted feed. The total trial duration was 38 days.

[0147] Despite vaccination, IBV still infected the birds, leading to an outbreak on day 20 in birds receiving a control diet. Surprisingly, in birds receiving the test diet, the disease broke out on day 29. Furthermore, compared to the control birds, the experimental birds showed significantly improved free radical saturation (FCR) and weight gain (1.58 vs. 1.69; and 2615 g vs. 2534 g, respectively). In addition, the mortality rate in the test diet was significantly lower than that in the control diet. Specifically, the mortality rate on days 23–35 was 14.48% in the control group and 2.965% in the experimental group. In contrast, there was no significant difference in mortality rate on days 0–10 between the two groups. Therefore, the oligosaccharide preparation was found to make birds more resilient and less susceptible to disease, and to delay and prolong IB outbreaks.

[0148] Example 2 For the individual feeding trials of laying hens, 24 replicate trials were conducted, each feeding eight Bovans white hens with either a control diet (containing phytase and non-starch polysaccharide xylanase), a first experimental diet (the control diet additionally containing 450 ppm of the oligosaccharide formulation of the present invention as described in Example 1), or a second experimental diet (the control diet additionally containing 900 ppm of the oligosaccharide formulation of the present invention as described in Example 1). All birds were vaccinated against IBV and NDV.

[0149] The experiment began when the birds were 22 weeks old and continued until they were 60 weeks old. Weekly monitoring included living conditions, feed consumption, egg production and weight, Haugh units, yolk color, eggshell crack strength, fertilizer moisture content, and mortality.

[0150] In terms of egg production, egg quality, and FCR, birds receiving any of the test diets outperformed those receiving the control diet throughout the trial. Outbreaks of IB and ND occurred at 26 weeks of age. Another ND outbreak was recorded at 36 weeks of age. Consistent with the findings of Example 1, disease outbreaks had a smaller impact on birds receiving any of the test diets compared to the control group.

[0151] At 22–25 weeks of age, compared with the control group, birds receiving 450 ppm and 900 ppm oligosaccharide formulations had average daily egg production (HDEP) that were 1.39% and 4.13% higher, respectively. This increased productivity was even more pronounced at 26–30 weeks (i.e., during disease outbreaks), with birds receiving 450 ppm and 900 ppm oligosaccharide formulations showing HDEP that were 7.66% and 11.34% higher, respectively, compared with birds receiving the control diet. Furthermore, in terms of average egg weight, birds receiving either test diet had an average egg weight (g / d) that was at least 2.2% higher than the control birds. Again, this improvement was particularly significant at 26–30 weeks, with birds receiving 450 ppm or 900 ppm of the present invention's oligosaccharide formulation producing an average egg weight that was 12.2% and 11.9% higher, respectively, than birds receiving the control diet. Similarly, regarding average free clotting rate (FCR), at 22-26 weeks of age, the experimental birds were 1-1.5% lower than the control birds (i.e., more efficient); at 26-30 weeks, 9.9-11.2% lower; at 30-34 weeks, 2.8-6.3% lower; and at 34-38 weeks, 1.6-4.9% lower, etc. Furthermore, it was found that the average shell cracking strength of eggs produced by the experimental birds was about 1% higher in the weeks preceding the disease outbreak, and about 3-11% higher than the control birds during or in the weeks following the disease outbreak. Notably, pO2 and pCO2, as blood biomarkers, indicated that the experimental birds experienced less respiratory stress than the control birds during the disease outbreak.

Claims

1. The use of oligosaccharide preparations for the prevention and / or delay of respiratory diseases, preferably infectious bronchitis, and / or their effects; The oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having a different degree of polymerization selected from 1 to n; wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

2. The use of oligosaccharide preparations to improve the efficacy of vaccines against respiratory diseases, preferably infectious bronchitis; The oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having a different degree of polymerization selected from 1 to n; wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

3. Use of oligosaccharide preparations for reducing, preventing and / or delaying mortality in poultry populations challenged by respiratory diseases, preferably infectious bronchitis; The oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having a different degree of polymerization selected from 1 to n; wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

4. The use of oligosaccharide preparations to reduce, alleviate, prevent and / or delay the incidence of respiratory diseases, preferably infectious bronchitis; The oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having a different degree of polymerization selected from 1 to n; wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

5. An oligosaccharide preparation for use in methods of treating, alleviating, preventing and / or delaying respiratory diseases, preferably infectious bronchitis and / or their effects; The oligosaccharide formulation comprises at least n oligosaccharide fractions (DP1 to DPn fractions) each having a different degree of polymerization selected from 1 to n; wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90%, for example 1% to 90%, or for example about 0.5% to about 15% of oligosaccharides containing dehydrated subunits, based on a relative abundance determined by mass spectrometry.

6. The oligosaccharide preparation according to claim 5 for use in a method of treating, alleviating, preventing and / or delaying respiratory diseases, preferably infectious bronchitis and / or their effects, wherein the oligosaccharide preparation is administered to an individual who has received a vaccine against the respiratory disease.

7. The use according to any one of claims 1-4; or the oligosaccharide preparation used according to any one of claims 5-6, wherein the oligosaccharide preparation is included in the nutritional composition at a concentration of greater than 40 ppm.

8. The use according to any one of claims 1-4, 7; or the oligosaccharide formulation used according to any one of claims 5-7, wherein n is at least 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, 4 4, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100.

9. The use according to any one of claims 1-4, 7-8; or the oligosaccharide formulation used according to any one of claims 5-8, wherein at least one fraction of the oligosaccharide formulation comprises less than 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% of oligosaccharides containing dehydrated subunits in relative abundance; and / or wherein each fraction of the oligosaccharide formulation comprises more than 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, 50%, 60%, 70% or 80% of oligosaccharides containing dehydrated subunits in relative abundance.

10. The use according to any one of claims 1-4, 7-9; or the oligosaccharide formulation used according to any one of claims 5-9, wherein the weight-average molecular weight of the oligosaccharide formulation is about 300 to 5000 g / mol, 500 to 5000 g / mol, 700 to 5000 g / mol, 500 to 2000 g / mol, 700 to 2000 g / mol, 700 to 1500 g / mol, 300 to 1500 g / mol, 300 to 2000 g / mol, 400 to 1300 g / mol, 400 to 1200 g / mol, 400 to 1100 g / mol, 500 to 1300 g / mol, 500 to 1200 g / mol, 500 to 1100 g / mol, 600 to 1300 g / mol, 600 to 1200 g / mol or 600 to 1100 g / mol; and / or the number average molecular weight of said oligosaccharide preparation is about 1000 to 2000 g / mol, 1100 to 1900 g / mol, 1200 to 1800 g / mol, 1300 to 1700 g / mol, 1400 to 1600 g / mol or 1450 to 1550 g / mol.

11. The use according to any one of claims 1-4, 7-10; or the oligosaccharide formulation used according to any one of claims 5-10, wherein the relative abundance of oligosaccharides in each of the n fractions of the oligosaccharide formulation decreases monotonically with its degree of polymerization.

12. The use according to any one of claims 1-4, 7-11; or the oligosaccharide formulation used according to any one of claims 5-11, wherein the average DP of the oligosaccharide formulation is at least 2, and / or wherein the average DP of the oligosaccharide formulation is less than 5.0.