Reducing effects of avian necrotizing enteritis using oligosaccharide preparations

Abstract

The present invention relates to a method of reducing the effect of necrotic enteritis in an avian comprising administering an oligosaccharide preparation, and an oligosaccharide preparation for use in treating, ameliorating and / or preventing an avian having or at risk of necrotic enteritis.

Description

[0001] The present invention relates to a method for reducing the effects of necrotic enteritis in poultry and the use of an oligosaccharide preparation.

[0002] Necrotic enteritis is one of the most common diseases in poultry farming, with severe consequences for the affected animals and economic losses for agricultural workers. Necrotic enteritis usually occurs in broilers, but can also be seen in other poultry species, such as laying hens or turkeys. In the event of a clinical outbreak, necrotic enteritis can cause up to 50% mortality. In addition, necrotic enteritis often causes intestinal lesions, which have negative effects on animal welfare and animal performance, such as reduced weight gain or increased feed conversion rate. It is estimated that necrotic enteritis causes economic losses of up to US$ 5 to 6 billion per year.

[0003] Necrotic enteritis is caused by an atypically high growth of Clostridium perfringens in the intestine. Under normal circumstances, C. perfringens is a regular member of the gut microbiota of birds. However, under certain conditions, C. perfringens can overproliferate and produce toxins. Conditions known to favor this overproliferation include consumption of feed contaminated with mycotoxins, coccidiosis stress caused by Eimeria contamination and / or vaccination, immunosuppression, low-quality protein, reduced amino acid digestibility or nutrient absorption, increased luminal protein availability, reduced gut integrity, protein leakage or pathogen translocation.

[0004] Traditionally, necrotic enteritis caused by Clostridium perfringens can be treated by administering antibiotics. However, in order to avoid the spread of bacterial mechanisms towards antibiotic resistance, the use of antibiotics in animal feed should be avoided as much as possible. Alternative measures to reduce the risk of necrotic enteritis outbreaks include fine-tuning feed composition and mode of administration. Despite these efforts, there is still a need for further measures to support the mitigation of the negative effects of necrotic enteritis. Therefore, in view of the above-mentioned prior art, the object of the present invention is to provide such methods and measures.

[0005] The object of the present invention is achieved by a method for reducing the effects of necrotic enteritis in poultry, the method comprising the step of administering an oligosaccharide preparation to poultry in need of reducing the effects of necrotic enteritis, wherein the oligosaccharide preparation comprises at least n oligosaccharide fractions, each oligosaccharide fraction 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% (e.g., 1% to 90%; or, for example, about 0.5% to about 15%) of oligosaccharides containing anhydro subunits in relative abundance as determined by mass spectrometry. The at least n fractions of oligosaccharides each having a different degree of polymerization selected from 1 to n can be referred to as DP1 to DPn fractions. By providing this method, one or more effects caused by necrotic enteritis can be reduced, mitigated or avoided. In certain embodiments, the effects of such necrotic enteritis are partially reduced. In other embodiments, the effects of such necrotic enteritis can be completely eliminated, i.e., poultry in need of reducing the effects of necrotic enteritis cannot be distinguished from poultry in need of reducing the effects of necrotic enteritis.

[0006] In another aspect, the present invention relates to the use of an oligosaccharide preparation for reducing the effects of necrotic enteritis in poultry in need thereof, wherein the oligosaccharide preparation comprises at least n oligosaccharide fractions, each oligosaccharide fraction having a different degree of polymerization selected from 1 to n (DP1 fraction to DPn fraction), wherein n is an integer greater than or equal to 2, and wherein each fraction comprises at least about 0.5% to about 90% (e.g., 1% to 90%; or, for example, about 0.5% to about 15%) of oligosaccharides containing anhydro subunits in relative abundance as determined by mass spectrometry. In particular, the oligosaccharide preparation is used for administration to poultry in need of reducing the effects of necrotic enteritis.

[0007] In another aspect, the present invention relates to an oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis, wherein the oligosaccharide preparation comprises at least n oligosaccharide fractions, each oligosaccharide fraction having a different degree of polymerization selected from 1 to n (DP1 fraction to DPn fraction), wherein n is an integer greater than or equal to 2, and wherein each fraction comprises at least about 0.5% to about 90% (e.g., 1% to 90%; or, for example, about 0.5% to about 15%) of anhydro subunit-containing oligosaccharides in relative abundance as determined by mass spectrometry.

[0008] In some embodiments, the effect of necrotic enteritis is at least one selected from the following groups: reduced weight gain in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds not requiring reduction of the effect of necrotic enteritis; increased feed conversion rate in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds; increased mortality (particularly increased mortality associated with necrotic enteritis) in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds; and increased lesion scores in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds.

[0009] In some embodiments, poultry in need of reducing the effects of necrotic enteritis are challenged with Clostridium perfringens. Such challenge with Clostridium perfringens includes an atypically high incidence of intestinal Clostridium perfringens. In some embodiments, the challenge with Clostridium perfringens due to overgrowth of Clostridium perfringens is the result of one or more of the following: consumption of feed contaminated with mycotoxins, coccidiosis stress caused by Eimeria contamination and / or vaccination against coccidiosis, immunosuppression, ingestion of low quality protein, reduced amino acid digestibility or nutrient absorption, increased luminal protein availability, reduced intestinal integrity, protein leakage or pathogen translocation; resulting in the need to reduce the effects of necrotic enteritis.

[0010] In some embodiments, the bird is poultry (e.g., chicken, duck, goose, turkey, guinea fowl, pigeon, quail). In some specific embodiments, the bird is a chicken, preferably a broiler chicken.

[0011] In some embodiments, the effects of necrotic enteritis are reduced on at least one day within 42 days after the first administration of the oligosaccharide preparation (on one or more days selected from the group consisting of 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days after administration of the oligosaccharide preparation). In some specific embodiments, the effects of necrotic enteritis are reduced compared to the day (i.e., day 1) on which the oligosaccharide preparation is first administered to the bird in need of reduced effects of necrotic enteritis. For example, a reduction in the effects of necrotic enteritis may be observed by comparing day 14 to day 1, day 21 to day 1, day 35 to day 1, and / or day 42 to day 1. Herein, "day 14" means the 14th day after the first administration of the oligosaccharide preparation to the poultry in need of a reduction in the effects of necrotic enteritis (i.e., after day 1); "day 21" means the 21st day after the first administration of the oligosaccharide preparation to the poultry in need of a reduction in the effects of necrotic enteritis (i.e., after day 1); and so on.

[0012] In some embodiments, the oligosaccharide preparation is included in a nutritional composition for administration to poultry.

[0013] In some embodiments, the oligosaccharide preparation is included in the nutritional composition at an inclusion rate of at least 50 ppm (e.g., at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm). In some specific embodiments, the oligosaccharide preparation is included in the nutritional composition at a concentration of at least 50 ppm (e.g., at least 100 ppm, 150 ppm, 200 ppm, 220 ppm, 250 ppm, 300 ppm, 310 ppm, 350 ppm, 400 ppm) of dry polysaccharides of the oligosaccharide preparation.

[0014] In some embodiments, the oligosaccharide preparation is administered to birds before, after and / or simultaneously with the diet of birds in need of reducing the effects of necrotic enteritis, preferably simultaneously with the diet.

[0015] The steps described herein of administering an oligosaccharide preparation to a poultry in need of reducing the effects of necrotic enteritis are intended to be interpreted as providing the oligosaccharide preparation to the poultry (e.g., as part of a nutritional composition) for consumption by the poultry. In other words, the oligosaccharide preparation is fed to the poultry. In one embodiment, such administration / providing / feeding can be achieved by providing the nutritional composition to the poultry in an ad libitum manner.

[0016] In some embodiments, administering the oligosaccharide preparation comprises providing the oligosaccharide preparation to the bird such that the bird can ingest the oligosaccharide preparation ad libitum. In these embodiments, the animal ingests at least a portion of the oligosaccharide preparation.

[0017] Furthermore, the oligosaccharide preparation may be provided to the bird on any suitable schedule. In some embodiments, the oligosaccharide preparation is provided to the bird daily, weekly, monthly, every other day, at least three days per week, or at least seven days per month.

[0018] In some embodiments, the oligosaccharide preparation is administered to the poultry multiple times a day. For example, in some embodiments, the oligosaccharide preparation is administered to the poultry at least once, twice, three times, four times, five times, six times, seven times, eight times, nine times, or ten times a day. In some embodiments, the oligosaccharide preparation is administered to the poultry at most once, twice, three times, four times, five times, six times, seven times, eight times, nine times, or ten times a day. In some embodiments, the oligosaccharide preparation is administered to the poultry at least once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times, fifteen times, or twenty times a week. In some embodiments, the oligosaccharide preparation is administered to the poultry at most once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times, fifteen times, or twenty times a week. In some embodiments, the oligosaccharide preparation is administered to the bird daily, every other day, every 3 days, every 4 days, weekly, every other week, or monthly.

[0019] In certain embodiments, the oligosaccharide preparation is administered to the bird at a certain time during the day. For example, in certain embodiments, the oligosaccharide preparation is administered to the bird in the morning, afternoon and / or evening, or any combination thereof.

[0020] In some embodiments, the oligosaccharide preparation is provided to the poultry at certain dietary stages. For example, a brooding diet is provided to some poultry between 0 days and 14 days of age. In other embodiments, for example, a growing diet is provided to poultry between 15 days and 28 days of age, between 15 days and 35 days of age, or between 15 days and 39 days of age. In still other embodiments, for example, a fattening diet is provided to poultry between 29 days and 35 days of age, between 36 days and 42 days of age, or between 40 days and 46 days of age.

[0021] The oligosaccharide preparation can be fed to individual animals or groups of animals. For example, in some embodiments, where the avian species is poultry, the oligosaccharide preparation can be fed to individual poultry or groups of poultry.

[0022] The oligosaccharide preparation can be provided to poultry in any suitable form, including, for example, solid form, liquid form, or a combination thereof. In addition, if the oligosaccharide preparation is provided as a part of a nutritional composition, the nutritional composition can be provided in any suitable form, including, for example, solid form, liquid form, or a combination thereof. In certain embodiments, the nutritional composition is a liquid, such as a syrup or a solution. In other embodiments, the nutritional composition or the oligosaccharide preparation is a solid, such as a pill or a powder. In still other embodiments, the nutritional composition or the oligosaccharide preparation can be fed to poultry in both a liquid component and a solid component, for example, in a paste form.

[0023] In some embodiments of the present invention, the oligosaccharide preparation referred to herein is a synthetic oligosaccharide preparation.

[0024] It is believed that the oligosaccharide preparation can be provided in the form of a powder formulation, which comprises at least 20% (w / w) (e.g. at least 20% w / w, 25% w / w, 30% w / w, 35% w / w, 40% w / w, 45% w / w, etc.) of an oligosaccharide preparation as mentioned herein; at least 25% (wt / wt) of a silica-based adsorbate (e.g. diatomaceous earth, amorphous precipitated silica) having an average particle size D of less than or equal to 3000 μm (e.g. 100-500 μm, 200-500 μm, 200-300 μm); and optionally 0-25% (w / w) of water and / or auxiliary substances; wherein the % is based on the total weight of the powder formulation. For example, such a powder formulation may contain 30-70% (w / w) of the oligosaccharide preparation mentioned herein; 30-70% (w / w) of a silica-based adsorbate (e.g., an average particle size of at least 50 μm); and 0-21% (w / w) of water; wherein the % is based on the total weight of the powder formulation. In some embodiments, the oligosaccharide preparation is formulated as described in any one of Examples 22 to 26 and 33 of WO 2020 / 097458.

[0025] In some embodiments, n of the oligosaccharide preparation 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.

[0026] In some embodiments, at least one fraction of the oligosaccharide preparation 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% anhydro subunit-containing oligosaccharides in relative abundance; and / or wherein each fraction of the oligosaccharide preparation 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 anhydro subunit-containing oligosaccharides in relative abundance.

[0027] In some embodiments, the weight average molecular weight of the oligosaccharide preparation is about 300 g / mol to 5000 g / mol (e.g., about 2000 g / mol to 2800 g / mol, 2100 g / mol to 2700 g / mol, 2200 g / mol to 2600 g / mol, 2300 g / mol to 2500 g / mol, or 2320 g / mol to 2420 g / mol), 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 2000 g / mol, 400 g / mol to 1300 g / mol. g / mol, 400 g / mol to 1200 g / mol, 400 g / mol to 1100 g / mol, 500 g / mol to 1300 g / mol, 500 g / mol to 1200 g / mol, 500 g / mol to 1100 g / mol, 600 g / mol to 1300 g / mol, 600 g / mol to 1200 g / mol, or 600 g / mol to 1100 g / mol; and / or wherein the number average molecular weight of the oligosaccharide preparation is about 1000 g / mol to 2000 g / mol, 1100 g / mol to 1900 g / mol, 1200 g / mol to 1800 g / mol, 1300 g / mol to 1700 g / mol, 1400 g / mol to 1600 g / mol, or 1450 g / mol to 1550 g / mol.

[0028] In some embodiments, the relative abundance of oligosaccharides in each of the n fractions of the oligosaccharide preparation decreases monotonically with its degree of polymerization.

[0029] The description and method for making the oligosaccharide preparation according to the invention are described in WO 2020 / 097458 and WO 2016 / 007778, in particular in the embodiments described therein, in particular any one of Examples 1 to 7, 16 to 18 of WO 2020 / 097458A1, in the method described in paragraph

[317] , and / or any one of Examples 73 to 77, 80 to 89, 97 to 99, 101 to 110 of WO 2016 / 007778 A1. The oligosaccharide preparation according to the invention is characterized in that an aqueous composition comprising one or more feed sugars and a catalyst is heated to a temperature and time sufficient to induce polymerization. Such oligosaccharide preparations may be referred to as synthetic oligosaccharide preparations. The oligosaccharide preparations as referred to herein are described in further detail below.

[0030] For the sake of clarity only, the term "oligosaccharide preparation" may refer to a preparation comprising one or more oligosaccharides.

[0031] As used herein, "oligosaccharide" or "oligomer" may refer to a monosaccharide, or a compound containing two or more monosaccharide subunits linked by glycosidic bonds. "Oligosaccharide" may also refer to an anhydromonosaccharide; or a compound containing two or more monosaccharide subunits, wherein at least one monosaccharide unit is replaced by an anhydro subunit. "Oligosaccharides" may optionally be functionalized. As used herein, the term oligosaccharide encompasses all kinds of oligosaccharides, wherein each monosaccharide subunit in the oligosaccharide is independently and optionally functionalized and / or replaced by its corresponding anhydromonosaccharide subunit.

[0032] "Anhydro subunit" can be a reversible thermal dehydration product of a monosaccharide (or monosaccharide subunit) or a sugar caramelization product. For example, the "anhydro subunit" can be an anhydromonosaccharide, such as anhydroglucose. As another example, the "anhydro subunit" can be connected to one or more conventional or anhydromonosaccharide subunits via a glycosidic bond.

[0033] Oligosaccharides may be characterized by containing two or more monosaccharide subunits linked by glycosidic bonds. In this regard, the term "oligoglucose" may refer to glucose or a compound containing two or more glucose monosaccharide subunits linked by glycosidic bonds. "Oligoglucose" may also refer to anhydroglucose; or a compound containing two or more glucose monosaccharide subunits linked by glycosidic bonds, wherein at least one monosaccharide subunit is replaced by anhydroglucose subunit. Similarly, "galacto-oligosaccharide" may refer to galactose; or a compound containing two or more galactose monosaccharide subunits linked by glycosidic bonds. "Galacto-oligosaccharide" may also refer to anhydrogalactose; or a compound containing two or more galactose monosaccharide subunits linked by glycosidic bonds, wherein at least one monosaccharide subunit is replaced by anhydrogalactose subunit. Similarly, oligoglucose-galactose can be oligoglucose, galacto-oligosaccharide or contain the compound of one or more glucose monosaccharide subunits and one or more galactose monosaccharide subunits connected by glycosidic bond, wherein at least one monosaccharide subunit in the monosaccharide subunit is replaced by its corresponding dehydrated monosaccharide subunit. Oligoglucose-galactose-xylose can refer to the compound produced by the condensation reaction of glucose, galactose and xylose. The oligosaccharide preparation comprising oligoglucose-galactose-xylose can comprise oligoglucose-galactose, oligoglucose-xylose, galacto-oligosaccharide-xylose, and contain the compound of one or more glucose monosaccharide subunits, one or more xylose monosaccharide subunits and one or more galactose monosaccharide subunits connected by glycosidic bond.

[0034] As used herein, unless otherwise indicated, the terms "monosaccharide unit" and "monosaccharide subunit" are used interchangeably. "Monosaccharide subunit" may refer to a monosaccharide monomer in an oligosaccharide. For an oligosaccharide with a degree of polymerization of 1, the oligosaccharide may be referred to as a monosaccharide subunit or a monosaccharide. For an oligosaccharide with a degree of polymerization greater than 1, its monosaccharide subunits are connected via glycosidic bonds.

[0035] As used herein, the term "conventional monosaccharide" may refer to a monosaccharide that does not contain an anhydro subunit. The term "conventional disaccharide" may refer to a disaccharide that does not contain an anhydro subunit. Therefore, the term "conventional subunit" may refer to a subunit that is not an anhydro subunit.

[0036] As used herein, the term "relative abundance" or "abundance" may refer to the abundance of a substance in terms of how common or rare it is. For example, a DP1 fraction containing 10% anhydro subunit-containing oligosaccharides in relative abundance may refer to a plurality of DP1 oligosaccharides, wherein 10% by number of the DP1 oligosaccharides are anhydromonosaccharides.

[0037] Degree of Polymerization (DP) Distribution:The degree of polymerization distribution of the oligosaccharide preparation can be determined by any suitable analytical method and instrument, including but not limited to the end group method, osmotic pressure method (osmotic pressure determination method), ultracentrifugation, viscosity measurement, light scattering method, size exclusion chromatography (SEC), SEC-MALLS, field flow fractionation (field flow fractionation, FFF), asymmetric flow field flow fractionation (asymmetric flow field flow fractionation, A4F), high-performance liquid chromatography (high-performance liquid chromatography, HPLC) and mass spectrometry (mass spectrometry, MS). For example, the distribution of the degree of polymerization can be determined and / or detected by mass spectrometry (e.g., MALDI-MS, LC-MS or GC-MS). For another example, the degree of polymerization distribution can be determined and / or detected by SEC, such as gel permeation chromatography (gel permeation chromatography, GPC). As another example, the degree of polymerization distribution can be determined and / or detected by HPLC, FFF or A4F. In another example, the degree of polymerization of the oligosaccharide preparation can be determined based on its molecular weight and molecular weight distribution (for a more detailed description, see WO 2020 / 097458).

[0038] Dehydrated subunit level: In some embodiments, each oligosaccharide fraction in the n oligosaccharide fractions of the oligosaccharide preparations described herein independently comprises a dehydration subunit level. For example, in some embodiments, the DP1 fraction comprises 10% of oligosaccharides containing dehydration subunits in relative abundance, and the DP2 fraction comprises 15% of oligosaccharides containing dehydration subunits in relative abundance. For another example, in some embodiments, the DP1 fraction, the DP2 fraction, and the DP3 fraction each comprise 5%, 10%, and 2% of oligosaccharides containing dehydration subunits in relative abundance, respectively. In other embodiments, two or more oligosaccharide fractions may comprise oligosaccharides containing dehydration subunits at similar levels. For example, in some embodiments, the DP1 fraction and the DP3 fraction each comprise about 5% of oligosaccharides containing dehydration subunits in relative abundance.

[0039] The level of dehydrated subunits 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 dehydrated subunits is at least partially determined by mass spectrometry such as MALDI-MS. In some embodiments, the level of dehydrated subunits can be at least partially determined by NMR. In some embodiments, the level of dehydrated subunits can be at least partially determined by HPLC. For example, in some embodiments, the level of dehydrated subunits can be determined by MALDI-MS, as described in more detail in WO 2020 / 097458.

[0040] Glycosidic bond :In some embodiments, the oligosaccharide preparations described herein include a variety of glycosidic bonds. The type and distribution of glycosidic bonds can depend on the source and manufacturing method of the oligosaccharide preparation. In some embodiments, the type and distribution of various glycosidic bonds can be measured and / or detected by any suitable method known in the art (e.g., NMR). For example, in some embodiments, glycosidic bonds are measured 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 at least partially measured and / or detected by proton NMR. In some embodiments, glycosidic bonds are at least partially measured and / or detected by carbon NMR. In some embodiments, glycosidic bonds are at least partially measured and / or detected by 2D HSQC NMR.

[0041] In some embodiments, the oligosaccharide preparation may contain 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.

[0042] In some embodiments, the distribution of glycosidic bond types of the oligosaccharide preparation is about 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%, % to 45 mol%, 15 mol% to 40 mol%, 15 mol% to 35 mol%, 20 mol% to 60 mol%, 20 mol% to 55 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%, 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% of α-(1,6) glycosidic bonds.

[0043] Molecular weight :The molecular weight and molecular weight distribution of oligosaccharide preparations can be determined by any suitable analytical means and instrument, such as end group method, osmotic pressure (osmotic pressure determination method), ultracentrifugation, viscosity measurement, light scattering method, SEC, SEC-MALLS, FFF, A4F, HPLC and mass spectrometry. In some embodiments, molecular weight and molecular weight distribution are measured by mass spectrometry, such as MALDI-MS, LC-MS or GC-MS. In some embodiments, molecular weight and molecular weight distribution are measured by size exclusion chromatography (SEC), such as gel permeation chromatography (GPC). In other embodiments, molecular weight and molecular weight distribution are measured by HPLC. In some embodiments, molecular weight and molecular weight distribution are measured by MALDI-MS.

[0044] In some embodiments, the weight average molecular weight of the oligosaccharide preparation is about 100 g / mol to 10000 g / mol, 200 g / mol to 8000 g / mol, 300 g / mol to 5000 g / mol, 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 900 g / mol to 5000 g / mol, 1100 g / mol to 5000 g / mol, 1300 g / mol to 5000 g / mol, 1500 g / mol to 5000 g / mol, 1700 g / mol to 5000 g / mol, 300 g / mol to 4500 g / mol, 500 g / mol to 4500 g / mol, g / mol, 700 g / mol to 4500 g / mol, 900 g / mol to 4500 g / mol, 1100 g / mol to 4500 g / mol, 1300 g / mol to 4500 g / mol, 1500 g / mol to 4500 g / mol, 1700 g / mol to 4500 g / mol, 1900 g / mol to 4500 g / mol, 300 g / mol to 4000 g / mol, 500 g / mol to 4000 g / mol, 700 g / mol to 4000 g / mol, 900 g / mol to 4000 g / mol, 1100 g / mol to 4000 g / mol, 1300 g / mol to 4500 g / mol, g / mol to 3000 g / mol, 1300 g / mol to 3000 g / mol, 1500 g / mol to 3000 g / mol, 1700 g / mol to 4000 g / mol, 1900 g / mol to 4000 g / mol, 300 g / mol to 3000 g / mol, 500 g / mol to 3000 g / mol, 700 g / mol to 3000 g / mol, 900 g / mol to 3000 g / mol, 1100 g / mol to 3000 g / mol, 1300 g / mol to 3000 g / mol, 1500 g / mol to 3000 g / mol, 1700 g / mol to 3000 g / mol, 1900 g / mol to 3000 g / mol, 2100 g / mol to 3000 g / mol g / mol to 3000 g / mol, 300 g / mol to 2500 g / mol, 500 g / mol to 2500 g / mol, 700 g / mol to 2500 g / mol, 900 g / mol to 2500 g / mol, 1100 g / mol to 2500 g / mol, 1300 g / mol to 2500 g / mol, 1500 g / mol to 2500 g / mol, 1700 g / mol to 2500 g / mol, 1900 g / mol to 2500 g / mol, 2100 g / mol to 2500 g / mol, 300 g / mol to 1500 g / mol, 500 g / mol to 1500 g / mol,In some embodiments, the weight average molecular weight of the oligosaccharide preparation is about 2000 g / mol to 2800 g / mol, 2100 g / mol to 2700 g / mol, 2200 g / mol to 2600 g / mol, 2300 g / mol to 2500 g / mol, or 2320 g / mol to 2420 g / mol.

[0045] Types of Oligosaccharides :In some embodiments, the type of oligosaccharide present in the oligosaccharide preparation mentioned herein may depend on the type of one or more feed sugars. For example, in some embodiments, when the feed sugar includes glucose, the oligosaccharide preparation includes oligoglucose. For example, in some embodiments, when the feed sugar includes galactose, the oligosaccharide preparation includes galacto-oligosaccharide. For another example, in some embodiments, when the feed sugar includes galactose and glucose, the oligosaccharide preparation includes oligoglucose-galactose.

[0046] In some embodiments, the oligosaccharide preparation comprises one or more types of monosaccharide subunits. In some embodiments, the oligosaccharide preparation can 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 types of monosaccharide subunits.

[0047] Method for producing oligosaccharide preparation: WO 2020 / 097458 describes in detail a method for producing an oligosaccharide preparation according to the present invention, the method 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 acid; butylphosphonic acid; diphenylphosphinic acid; hexylphosphonic acid; methylphosphonic acid; phenylphosphinic acid; phenylphosphonic acid; tert-butylphosphonic acid; SS)-VAPOL hydrogen phosphate; 6-quinolinesulfonic acid, 3-(1-pyridyl)-1-propanesulfonic acid; 2-(2-pyridyl)ethanesulfonic acid; 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine-p,p'-disulfonic acid monosodium salt hydrate; 1,1'-binaphthyl-2,2'-diyl-hydrogen phosphate; bis(4-methyl)-1,1'-diphenylphosphine; 1-Hydroxyphenyl)phosphinic acid; phenyl(3,5-xylyl)phosphinic acid; L-cysteic acid monohydrate; poly(styrenesulfonic acid-co-divinylbenzene); lysine; ethanedisulfonic acid; ethanesulfonic acid; isethionic acid; homocysteine; HEPBS (N-(2-hydroxyethyl)piperazine-N'-(4-butanesulfonic acid)); HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid); 2-hydroxy-3-morpholinopropanesulfonic acid; 2-(N-morpholino)ethanesulfonic acid ; methanesulfonic acid; formoyl hydrazide; naphthalene-1-sulfonic acid; naphthalene-2-sulfonic acid; perfluorobutanesulfonic acid; 6-sulfoquinovose; 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.

[0048] In some embodiments, polymerization of the feed sugar is achieved by step-growth polymerization. In some embodiments, polymerization of the feed sugar is achieved by condensation polymerization.

[0049] Feed sugar: The one or more feed sugars used in the methods of making the oligosaccharide preparations 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.

[0050] In some embodiments, the one or more feed sugars include glucose. In some embodiments, the one or more feed sugars include glucose and galactose. In some embodiments, the one or more feed sugars include glucose, xylose and galactose. In some embodiments, the one or more feed sugars include glucose and mannose. In some embodiments, the one or more feed sugars include glucose and fructose. In some embodiments, the one or more feed sugars include glucose, fructose and galactose. In some embodiments, the one or more feed sugars include glucose, galactose and mannose.

[0051] As used herein, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "an agent" includes a plurality of such agents, reference to "the oligosaccharide" includes reference to one or more oligosaccharides (or oligosaccharides) and equivalents thereof known to those skilled in the art, and so forth.

[0052] When ranges are used herein for physical properties (e.g., molecular weight) or chemical properties (e.g., chemical formula), all combinations and subcombinations of ranges and specific embodiments therein are intended to be included. When referring to a number or a numerical range, the term "about" means that the number or numerical range referred to is an approximation within experimental variability (or statistical experimental error), and thus in some cases, the number or numerical range will vary between 1% and 15% of the number or numerical range.

[0053] The present invention is further characterized by the following items:

[0054] Item 1: A method for reducing the effects of necrotic enteritis in poultry, comprising the step of administering an oligosaccharide preparation to poultry in need of reducing the effects of necrotic enteritis, wherein the oligosaccharide preparation comprises at least n oligosaccharide fractions, each oligosaccharide fraction having a different degree of polymerization selected from 1 to n (DP1 fraction to DPn fraction), wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90% (e.g., 1% to 90%; or, for example, about 0.5% to about 15%) of oligosaccharides containing anhydro subunits, in relative abundance as determined by mass spectrometry.

[0055] Item 2: The method according to item 1, wherein the effect of necrotic enteritis is at least one selected from the following groups: reduced weight gain in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds not requiring reduction of the effect of necrotic enteritis; increased feed conversion rate in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds; increased mortality in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds; and increased lesion score in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds.

[0056] Item 3: A method according to item 1 or 2, wherein the poultry in which the effects of necrotic enteritis need to be reduced are challenged with Clostridium perfringens.

[0057] Item 4: Method according to any of the preceding items, wherein the oligosaccharide preparation is comprised in a nutritional composition for administration to poultry.

[0058] Item 5: A method according to any of the preceding items, wherein the oligosaccharide preparation is included in the nutritional composition at an inclusion rate of at least 50 ppm (e.g., at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm).

[0059] Item 6: Method according to any of the preceding items, wherein the oligosaccharide preparation is administered to birds before, after and / or simultaneously with the diet of birds in need of reducing the effects of necrotic enteritis, preferably simultaneously with the diet.

[0060] Item 7: The method according to any of the preceding items, wherein the bird is poultry (eg, chicken, duck, goose, turkey, guinea fowl, pigeon, quail); preferably a broiler chicken.

[0061] Item 8: A method according to any of the preceding items, wherein the effects of necrotic enteritis are reduced on at least one day within 42 days after the first administration of the oligosaccharide preparation (on one or more days selected from the following group: 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days after administration of the oligosaccharide preparation).

[0062] Item 9: A method according to any one of the preceding items, wherein n of the oligosaccharide preparation 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.

[0063] Item 10: A method according to any of the preceding items, wherein at least one fraction of the oligosaccharide preparation 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 anhydro subunit-containing and / or wherein each fraction of the oligosaccharide preparation 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 anhydro subunits in relative abundance.

[0064] Item 11: The method according to any one of the preceding items, wherein the weight average molecular weight of the oligosaccharide preparation is about 300 g / mol to 5000 g / mol (e.g., about 2000 g / mol to 2800 g / mol, 2100 g / mol to 2700 g / mol, 2200 g / mol to 2600 g / mol, 2300 g / mol to 2500 g / mol, or 2320 g / mol to 2420 g / mol), 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 2000 g / mol, 400 g / mol g / mol, 1100 g / mol, 1900 g / mol, 2000 g / mol, 2100 g / mol, 2200 g / mol, 2400 g / mol, 2500 g / mol, 2600 g / mol, 2700 g / mol, 2800 g / mol, 2900 g / mol, 3000 g / mol, 3100 g / mol, 3200 g / mol, 3300 g / mol, 3400 g / mol, 3600 g / mol, 3700 g / mol, 3800 g / mol, 3900 g / mol, 400 g / mol, 4100 g / mol, 4200 g / mol, 4300 g / mol, 4400 g / mol, 450 g / mol, 4600 g / mol, 4700 g / mol, 4800 g / mol, 4900 g / mol, 500 g / mol, 5100 g / mol, 5100 g / mol, 5200 g / mol, 5200 g / mol

[0065] Item 12: The method according to any one of the preceding items, wherein the relative abundance of oligosaccharides in each of the n fractions of the oligosaccharide preparation decreases monotonically with its degree of polymerization.

[0066] Item 13: Method according to any of the preceding items, wherein the relative abundance of the oligosaccharides in at least 5, 10, 20 or 30 DP fractions of the oligosaccharide preparation decreases monotonically with its degree of polymerization.

[0067] Item 14: A method according to any of the preceding items, wherein the oligosaccharide preparation 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% anhydro subunit-containing oligosaccharides in relative abundance.

[0068] Item 15: A method according to any of the preceding items, wherein each fraction of the oligosaccharide preparation contains 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 anhydro subunits in relative abundance.

[0069] Item 16: A method according to any of the preceding items, wherein at least one fraction of the oligosaccharide preparation comprises 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 oligosaccharides containing anhydro subunits in relative abundance.

[0070] Item 17: A method according to any of the preceding items, wherein the oligosaccharide preparation comprises 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 oligosaccharides containing anhydro subunits in relative abundance.

[0071] Item 18: The method according to any one of the preceding items, wherein each fraction of the oligosaccharide preparation comprises greater than 20%, 21%, 22%, 23%, 24% or 25% of anhydro subunit-containing oligosaccharides in relative abundance.

[0072] Item 19: A method according to any of the preceding items, wherein greater than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35% or 30% of the anhydrosubunit-containing oligosaccharides of the oligosaccharide preparation have only one anhydrosubunit.

[0073] Item 20: The method according to any one of the preceding items, wherein the oligosaccharide preparation has a DP1 fraction content of 1% to 40% in relative abundance.

[0074] Item 21: Method according to any of the preceding items, wherein the oligosaccharide preparation has a DP2 fraction content of 1% to 35% in relative abundance.

[0075] Item 22: The method according to any one of the preceding items, wherein the oligosaccharide preparation has a DP3 fraction content of 1% to 30% in relative abundance.

[0076] Item 23: The method according to any one of the preceding items, wherein the oligosaccharide preparation has a DP4 fraction content of 0.1% to 20% in relative abundance.

[0077] Item 24: The method according to any one of the preceding items, wherein the oligosaccharide preparation has a DP5 fraction content of 0.1% to 15% in relative abundance.

[0078] Item 25: The method according to any one of the preceding items, wherein the ratio of the DP2 fraction to the DP1 fraction of the oligosaccharide preparation is 0.02-0.40 in relative abundance.

[0079] Item 26: The method according to any one of the preceding items, wherein the ratio of the DP3 fraction to the DP2 fraction of the oligosaccharide preparation is 0.01-0.30 in relative abundance.

[0080] Item 27: The method according to any one of the preceding items, wherein the aggregate content of the DP1 fraction and the DP2 fraction in the oligosaccharide preparation is less than 50%, 30% or 10% in relative abundance.

[0081] Item 28: The method according to any of the preceding items, wherein the oligosaccharide preparation comprises at least 103, 104, 105, 106 or 109 different oligosaccharide species.

[0082] Item 29: The method according to any one of the preceding items, wherein two or more individual oligosaccharides of the oligosaccharide preparation comprise different anhydro subunits.

[0083] Item 30: The method according to any one of the preceding items, wherein the oligosaccharide preparation comprises one or more anhydro subunits which are the product of reversible thermal dehydration of monosaccharides.

[0084] Item 31: A method according to any one of the preceding items, wherein the oligosaccharide preparation comprises one or more anhydroglucose, anhydrogalactose, anhydromannose, anhydroallose, anhydroaltrose, anhydrogulose, anhydroidose, anhydrotalose, anhydrofructose, anhydroribose, anhydroarabinose, anhydrorhamnose, anhydrolyxose or anhydroxylose subunits.

[0085] Item 32: The method according to any one of the preceding items, wherein the oligosaccharide preparation comprises one or more anhydroglucose, anhydrogalactose, anhydromannose or anhydrofructose subunits.

[0086] Item 33: The method according to any one of the preceding items, wherein the oligosaccharide preparation comprises one or more 1,6-anhydro-β-D-glucopyranose or 1,6-anhydro-β-D-glucopyranose subunits. In some embodiments, the oligosaccharide preparation comprises both 1,6-anhydro-β-D-glucopyranose and 1,6-anhydro-β-D-glucopyranose anhydro subunits.

[0087] Item 34: A method according to any one of the preceding items, wherein the ratio of 1,6-anhydro-β-D-furanose to 1,6-anhydro-β-D-pyranose in the oligosaccharide preparation 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.

[0088] Item 35: A method according to any of the preceding items, wherein the ratio of 1,6-anhydro-β-D-furanose to 1,6-anhydro-β-D-pyranose in the oligosaccharide preparation 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, 1:8, 1:9, or 1:10.

[0089] Item 36: The method according to any one of the preceding items, wherein the ratio of 1,6-anhydro-β-D-glucopyranose to 1,6-anhydro-β-D-glucopyranose in the oligosaccharide preparation is about 2:1.

[0090] Item 37: A method according to any of the preceding items, wherein the ratio of 1,6-anhydro-β-D-glucopyranose to 1,6-anhydro-β-D-glucopyranose in each fraction of the oligosaccharide preparation 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.

[0091] Item 38: A method according to any of the preceding items, wherein the ratio of 1,6-anhydro-β-D-furanose to 1,6-anhydro-β-D-pyranose in each fraction of the oligosaccharide preparation 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, 1:8, 1:9, or 1:10.

[0092] Item 39: The method according to any one of the preceding items, wherein the ratio of 1,6-anhydro-β-D-glucopyranose to 1,6-anhydro-β-D-glucopyranose in each fraction of the oligosaccharide preparation is about 2:1.

[0093] Item 40: A method according to any of the preceding items, wherein at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% of the anhydro subunits in the oligosaccharide preparation are selected from the group consisting of: 1,6-anhydro-β-D-furanose and 1,6-anhydro-β-D-pyranose.

[0094] Item 41: The method according to any one of the preceding items, wherein the weight average molecular weight of the oligosaccharide preparation is about 300 g / mol to 5000 g / mol, 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 2 In some embodiments, the present invention relates to a polyol having a molecular weight of 400 g / mol to 1000 g / mol, 400 g / mol to 1300 g / mol, 400 g / mol to 1200 g / mol, 400 g / mol to 1100 g / mol, 500 g / mol to 1300 g / mol, 500 g / mol to 1200 g / mol, 500 g / mol to 1100 g / mol, 600 g / mol to 1300 g / mol, 600 g / mol to 1200 g / mol, or 600 g / mol to 1100 g / mol.

[0095] Item 42: A method according to any of the preceding items, wherein the number average molecular weight of the oligosaccharide preparation is about 300 g / mol to 5000 g / mol, 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 2000 g / mol, 400 g / mol to 1000 g / mol, 400 g / mol to 900 g / mol, 400 g / mol to 800 g / mol, 500 g / mol to 900 g / mol, or 500 g / mol to 800 g / mol.

[0096] Item 43: The method according to any one of the preceding items, wherein the degree of polymerization distribution is determined and / or detected by MALDI-MS, GC-MS, LC-MS, SEC, HPLC and / or a combination thereof (e.g., MALDI-MS and SEC).

[0097] Item 44: The method according to any of the preceding items, wherein the degree of polymerization of the oligosaccharide preparation can be determined based on its molecular weight and molecular weight distribution.

[0098] Item 45: A method according to any of the preceding items, wherein the oligosaccharide preparation is contained in the nutritional composition at a concentration of at least 50 g / ton of feed (e.g. at least 70 g / ton of feed, 100 g / ton of feed, 200 g / ton of feed, 300 g / ton of feed, 400 g / ton of feed, 500 g / ton of feed, 600 g / ton of feed, 700 g / ton of feed, 800 g / ton of feed, 900 g / ton of feed); and / or wherein the oligosaccharide preparation is contained in the nutritional composition at an inclusion rate of at least 50 ppm (e.g. at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 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, 500 ppm).

[0099] Item 46: The method according to any of the preceding items, wherein the oligosaccharide preparation is administered for at least one day, preferably at least 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 days, 45 days, 46 days, 47 days, 48 ​​days, 49 days, 50 days, 51 days, 52 days, 53 days, 54 days, 55 days day, 56 days, 57 days, 58 days, 59 days, 60 days, 61 days, 62 days, 63 days, 64 days, 65 days, 66 days, 67 days, 68 days, 69 days, 70 days, 71 days, 72 days, 73 days, 74 days, 75 days, 76 days, 77 days, 78 days, 79 days, 80 days, 81 days, 82 days, 83 days, 84 days, 85 days, 86 days Days, 87 days, 88 days, 89 days, 90 days, 91 days, 92 days, 93 days, 94 days, 95 days, 96 days, 97 days, 98 days, 99 days, 100 days, 101 days, 102 days, 103 days, 104 days, 105 days, 106 days, 107 days, 108 days, 109 days, 110 days, 111 days, 112 days, 113 days, 114 days 14 days, 115 days, 116 days, 117 days, 118 days, 119 days, 120 days, 121 days, 122 days, 123 days, 124 days, 125 days, 126 days, 127 days, 128 days, 129 days, 130 days, 131 days, 132 days, 133 days, 134 days, 135 days, 136 days, 137 days, 138 days, 139 days, 140 days, 141 days, 142 days, 143 days, 144 days, 145 days, 146 days, 147 days, 148 days, 149 days, 150 days, 151 days, 152 days, 153 days, 154 days, 155 days, 156 days, 157 days, 158 days, 159 days, 160 days, 161 days, 162 days, 163 days, 164 days, 165 days, 166 days, 167 days, 168 days, 169 days, 170 days, 171 days, 172 days, 173 days, 174 days, 175 days, 176 days, 177 days, 178 days, 179 days, 180 days, 181 days, 182 days, 183 days, 184 days, 185 days, 186 days, 187 days, 188 days, 189 days, 190 days, 191 days, 192 days, 193 days, 194 days, 195 days, 196 days, 197 days, 198 days, 199 days, 200 days, 201 days, 202 days, 203 days, 204 days, 205 days, 206 days, 207 days, 208 days, 209 days, 210 days, 211 days, 212 days, 213 days,214 days, 215 days, 216 days, 217 days, 218 days, 219 days, 220 days, 221 days, 222 days, 223 days, 224 days, 225 days, 226 days, 227 days, 228 days, 229 days, 230 days, 231 days, 232 days, 233 days, 234 days, 235 days, 236 days, 237 days, 238 days, 239 days, 240 days, 241 days, 242 days, 243 days, 244 days or 245 days, most preferably continuously (i.e., uninterruptedly) administering the nutritional composition.

[0100] Item 47: Use of an oligosaccharide preparation for reducing the effects of necrotic enteritis in poultry in need thereof, wherein the oligosaccharide preparation comprises at least n oligosaccharide fractions, each oligosaccharide fraction having a different degree of polymerization selected from 1 to n (DP1 fraction to DPn fraction), wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90% (e.g., 1% to 90%; or, for example, about 0.5% to about 15%) of oligosaccharides containing anhydro subunits, in relative abundance as determined by mass spectrometry.

[0101] Item 48: The use according to Item 47, 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, 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 9, 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.

[0102] Item 49: Use according to item 47 or 48, wherein at least one fraction of the oligosaccharide preparation 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 anhydro subunit-containing oligosaccharide in relative abundance. Oligosaccharides; and / or wherein each fraction of the oligosaccharide preparation 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 anhydro subunits in relative abundance.

[0103] Item 50: The use according to any one of items 47 to 49, wherein the weight average molecular weight of the oligosaccharide preparation is about 300 g / mol to 5000 g / mol (e.g., about 2000 g / mol to 2800 g / mol, 2100 g / mol to 2700 g / mol, 2200 g / mol to 2600 g / mol, 2300 g / mol to 2500 g / mol, or 2320 g / mol to 2420 g / mol), 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 2000 g / mol, 400 g / mol to 5000 g / mol 1 to 1300 g / mol, 400 g / mol to 1200 g / mol, 400 g / mol to 1100 g / mol, 500 g / mol to 1300 g / mol, 500 g / mol to 1200 g / mol, 500 g / mol to 1100 g / mol, 600 g / mol to 1300 g / mol, 600 g / mol to 1200 g / mol, or 600 g / mol to 1100 g / mol; and / or wherein the number average molecular weight of the oligosaccharide preparation is about 1000 g / mol to 2000 g / mol, 1100 g / mol to 1900 g / mol, 1200 g / mol to 1800 g / mol, 1300 g / mol to 1700 g / mol, 1400 g / mol to 1600 g / mol, or 1450 g / mol to 1550 g / mol.

[0104] Item 51: Use according to any one of Items 47-50, wherein the relative abundance of oligosaccharides in each of said n fractions of said oligosaccharide preparation decreases monotonically with its degree of polymerization.

[0105] Item 52: Use according to any one of items 47-51, wherein the relative abundance of the oligosaccharides in at least 5, 10, 20 or 30 DP fractions of the oligosaccharide preparation decreases monotonically with its degree of polymerization.

[0106] Item 53: Use according to any one of items 47-52, wherein the oligosaccharide preparation 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% anhydro subunit-containing oligosaccharides in relative abundance.

[0107] Item 54: Use according to any one of items 47-53, wherein each fraction of the oligosaccharide preparation contains 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 anhydro subunits in relative abundance.

[0108] Item 55: Use according to any one of items 47-54, wherein at least one fraction of the oligosaccharide preparation comprises 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 oligosaccharides containing anhydro subunits in relative abundance.

[0109] Item 56: Use according to any one of items 47-55, wherein the oligosaccharide preparation comprises 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 oligosaccharides containing anhydro subunits in relative abundance.

[0110] Item 57: Use according to any one of items 47-56, wherein each fraction of the oligosaccharide preparation comprises greater than 20%, 21%, 22%, 23%, 24% or 25% of anhydro subunit-containing oligosaccharides in relative abundance.

[0111] Item 58: Use according to any one of items 47-57, wherein greater than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35% or 30% of the anhydrosubunit-containing oligosaccharides of the oligosaccharide preparation have only one anhydrosubunit.

[0112] Item 59: Use according to any one of items 47-58, wherein the oligosaccharide preparation has a DP1 fraction content of 1% to 40% in relative abundance.

[0113] Item 60: Use according to any one of items 47-59, wherein the oligosaccharide preparation has a DP2 fraction content of 1% to 35% in relative abundance.

[0114] Item 61: Use according to any one of items 47-60, wherein the oligosaccharide preparation has a DP3 fraction content of 1% to 30% in relative abundance.

[0115] Item 62: Use according to any one of items 47 to 61, wherein the oligosaccharide preparation has a DP4 fraction content of 0.1% to 20% in relative abundance.

[0116] Item 63: Use according to any one of items 47-62, wherein the oligosaccharide preparation has a DP5 fraction content of 0.1% to 15% in relative abundance.

[0117] Item 64: Use according to any one of items 47 to 63, wherein the ratio of the DP2 fraction to the DP1 fraction of the oligosaccharide preparation is 0.02 to 0.40 in relative abundance.

[0118] Item 65: Use according to any one of items 47 to 64, wherein the ratio of the DP3 fraction to the DP2 fraction of the oligosaccharide preparation is 0.01 to 0.30 in relative abundance.

[0119] Item 66: Use according to any one of items 47 to 65, wherein the aggregate content of the DP1 fraction and the DP2 fraction in the oligosaccharide preparation is less than 50%, 30% or 10% in relative abundance.

[0120] Item 67: Use according to any of items 47 to 66, wherein the oligosaccharide preparation comprises at least 103, 104, 105, 106 or 109 different oligosaccharide species.

[0121] Item 68: Use according to any one of items 47 to 67, wherein two or more separate oligosaccharides of the oligosaccharide preparation comprise different anhydro subunits.

[0122] Item 69: Use according to any one of items 47-68, wherein the oligosaccharide preparation comprises one or more anhydro subunits which are the product of reversible thermal dehydration of monosaccharides.

[0123] Item 70: Use according to any one of items 47 to 69, wherein the oligosaccharide preparation comprises one or more anhydroglucose, anhydrogalactose, anhydromannose, anhydroallose, anhydroaltrose, anhydrogulose, anhydroidose, anhydrotalose, anhydrofructose, anhydroribose, anhydroarabinose, anhydrorhamnose, anhydrolyxose or anhydroxylose subunits.

[0124] Item 71: Use according to any one of items 47 to 70, wherein the oligosaccharide preparation comprises one or more anhydroglucose, anhydrogalactose, anhydromannose or anhydrofructose subunits.

[0125] Item 72: The use according to any one of items 47 to 71, wherein the oligosaccharide preparation comprises one or more 1,6-anhydro-β-D-glucopyranose subunits or 1,6-anhydro-β-D-glucopyranose subunits. In some embodiments, the oligosaccharide preparation comprises both 1,6-anhydro-β-D-glucopyranose and 1,6-anhydro-β-D-glucopyranose anhydrosubunits.

[0126] Item 73: Use according to any one of items 47 to 72, wherein the ratio of 1,6-anhydro-β-D-glucopyranose to 1,6-anhydro-β-D-glucopyranose in the oligosaccharide preparation 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.

[0127] Item 74: Use according to any one of items 47 to 73, wherein the ratio of 1,6-anhydro-β-D-furanose to 1,6-anhydro-β-D-pyranose in the oligosaccharide preparation 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, 1:8, 1:9, or 1:10.

[0128] Item 75: Use according to any one of items 47 to 74, wherein the ratio of 1,6-anhydro-β-D-glucopyranose to 1,6-anhydro-β-D-glucopyranose in the oligosaccharide preparation is about 2:1.

[0129] Item 76: Use according to any one of items 47 to 75, wherein the ratio of 1,6-anhydro-β-D-glucopyranose to 1,6-anhydro-β-D-glucopyranose in each fraction of the oligosaccharide preparation 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.

[0130] Item 77: Use according to any one of items 47 to 76, wherein the ratio of 1,6-anhydro-β-D-furanose to 1,6-anhydro-β-D-pyranose in each fraction of the oligosaccharide preparation 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, 1:8, 1:9, or 1:10.

[0131] Item 78: Use according to any one of items 47 to 77, wherein the ratio of 1,6-anhydro-β-D-glucopyranose to 1,6-anhydro-β-D-glucopyranose in each fraction of the oligosaccharide preparation is about 2:1.

[0132] Item 79: Use according to any one of items 47-78, wherein at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% of the anhydro subunits in the oligosaccharide preparation are selected from the group consisting of: 1,6-anhydro-β-D-furanose and 1,6-anhydro-β-D-pyranose.

[0133] Item 80: The use according to any one of items 47 to 79, wherein the weight average molecular weight of the oligosaccharide preparation is about 300 g / mol to 5000 g / mol, 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, g / mol, 500 g / mol to 1300 g / mol, 500 g / mol to 1200 g / mol, 500 g / mol to 1100 g / mol, 600 g / mol to 1300 g / mol, 600 g / mol to 1200 g / mol, or 600 g / mol to 1100 g / mol.

[0134] Item 81: Use according to any one of items 47 to 80, wherein the number average molecular weight of the oligosaccharide preparation is about 300 g / mol to 5000 g / mol, 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 2000 g / mol, 400 g / mol to 1000 g / mol, 400 g / mol to 900 g / mol, 400 g / mol to 800 g / mol, 500 g / mol to 900 g / mol, or 500 g / mol to 800 g / mol.

[0135] Item 82: Use according to any one of items 47 to 81, wherein the degree of polymerization distribution is determined and / or detected by MALDI-MS, GC-MS, LC-MS, SEC, HPLC and / or a combination thereof (e.g. MALDI-MS and SEC).

[0136] Item 83: Use according to any one of items 47 to 84, wherein the degree of polymerization of the oligosaccharide preparation can be determined based on its molecular weight and molecular weight distribution.

[0137] Item 84: Use according to any one of items 47 to 83, wherein the oligosaccharide preparation is contained in the nutritional composition at a concentration of at least 50 g / ton of feed (e.g. at least 70 g / ton of feed, 100 g / ton of feed, 200 g / ton of feed, 300 g / ton of feed, 400 g / ton of feed, 500 g / ton of feed, 600 g / ton of feed, 700 g / ton of feed, 800 g / ton of feed, 900 g / ton of feed); and / or wherein the oligosaccharide preparation is contained in the nutritional composition at a inclusion rate of at least 50 ppm (e.g. at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 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, 500 ppm).

[0138] Item 85: Use according to any of items 47 to 84, wherein the oligosaccharide preparation is administered for at least one day, preferably at least 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 days, 45 days, 46 days, 47 days, 48 ​​days, 49 days, 50 days, 51 days, 52 days, 53 days, 54 days, 55 days, 56 days, 57 days, 58 days, 59 days, 60 days, 61 days, 62 days, 63 days, 64 days, 65 days, 66 days, 67 days, 68 days, 69 days, 70 days, 71 days, 72 days, 73 days, 74 days, 75 days, 76 days, 77 days, 78 days, 79 days, 80 days, 81 days, 82 days, 83 days, 84 days, 85 days, 86 days 6 days, 87 days, 88 days, 89 days, 90 days, 91 days, 92 days, 93 days, 94 days, 95 days, 96 days, 97 days, 98 days, 99 days, 100 days, 101 days, 102 days, 103 days, 104 days, 105 days, 106 days, 107 days, 108 days, 109 days, 110 days, 111 days, 112 days, 113 days, 114 days, 115 days, 116 days, 117 days, 118 days, 119 days, 120 days, 121 days, 122 days, 123 days, 124 days, 125 days, 126 days, 127 days, 128 days, 129 days, 130 days, 131 days, 132 days, 133 days, 134 days, 135 days, 136 days, 137 days, 138 days, 139 days, 140 days, 141 days, 142 days, 143 days, 144 days, 145 days, 146 days, 147 days, 148 days, 149 days, 150 days, 151 days, 152 days, 153 days, 154 days, 155 days, 156 days, 157 days, 158 days, 159 days, 160 days, 161 days, 162 days, 163 days, 164 days, 165 days, 166 days, 167 days, 168 days, 169 days, 170 days, 171 days, 172 days, 173 days, 174 days, 175 days, 176 days, 177 days, 178 days, 179 days, 180 days, 181 days, 182 days, 183 days, 184 days, 185 days, 186 days, 187 days, 188 days, 189 days, 190 days, 191 days, 192 days, 193 days, 194 days, 195 days, 196 days, 197 days, 198 days, 199 days, 200 days, 201 days, 202 days, 203 days, 204 days, 205 days, 206 days, 207 days, 208 days, 209 days, 210 days, 211 days, 212 days, 213 days,214 days, 215 days, 216 days, 217 days, 218 days, 219 days, 220 days, 221 days, 222 days, 223 days, 224 days, 225 days, 226 days, 227 days, 228 days, 229 days, 230 days, 231 days, 232 days, 233 days, 234 days, 235 days, 236 days, 237 days, 238 days, 239 days, 240 days, 241 days, 242 days, 243 days, 244 days or 245 days, most preferably continuously (i.e., uninterruptedly) administering the nutritional composition.

[0139] Item 86: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis, wherein the oligosaccharide preparation comprises at least n oligosaccharide fractions, each oligosaccharide fraction having a different degree of polymerization selected from 1 to n (DP1 fraction to DPn fraction), wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90% (e.g., 1% to 90%; or, for example, about 0.5% to about 15%) of oligosaccharides containing anhydro subunits, in relative abundance as determined by mass spectrometry.

[0140] Item 87: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to Item 86, wherein n of the oligosaccharide preparation 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 1, 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.

[0141] Item 88: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to item 86 or 87, wherein at least one fraction of the oligosaccharide preparation comprises less than 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17 ... and / or wherein each fraction of the oligosaccharide preparation 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 anhydro subunits in relative abundance.

[0142] Item 89: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 88, wherein the weight average molecular weight of the oligosaccharide preparation is about 300 g / mol to 5000 g / mol (e.g., about 2000 g / mol to 2800 g / mol, 2100 g / mol to 2700 g / mol, 2200 g / mol to 2600 g / mol). , 2300 g / mol to 2500 g / mol, or 2320 g / mol to 2420 g / mol), 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, ol to 2000g / mol, 400g / mol to 1300g / mol, 400g / mol to 1200g / mol, 400g / mol to 1100g / mol, 500g / mol to 1300g / mol, 500g / mol to 1200g / mol, 500g / mol to 1100g / mol, 600g / mol to 1300g / mol, 600g / mol to 1200g / mol 1, or 600 g / mol to 1100 g / mol; and / or wherein the number average molecular weight of the oligosaccharide preparation is about 1000 g / mol to 2000 g / mol, 1100 g / mol to 1900 g / mol, 1200 g / mol to 1800 g / mol, 1300 g / mol to 1700 g / mol, 1400 g / mol to 1600 g / mol, or 1450 g / mol to 1550 g / mol.

[0143] Item 90: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86-89, wherein the relative abundance of oligosaccharides in each of the n fractions of the oligosaccharide preparation decreases monotonically with its degree of polymerization.

[0144] Item 91: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86-90, wherein the relative abundance of the oligosaccharides in at least 5, 10, 20 or 30 DP fractions of the oligosaccharide preparation decreases monotonically with its degree of polymerization.

[0145] Item 92: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86-91, wherein the oligosaccharide preparation 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 anhydro subunit-containing oligosaccharides in relative abundance.

[0146] Item 93: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86-92, wherein each fraction of the oligosaccharide preparation 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 anhydro subunits in relative abundance.

[0147] Item 94: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86-93, wherein at least one fraction of the oligosaccharide preparation comprises 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 anhydro subunit-containing oligosaccharides in relative abundance.

[0148] Item 95: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86-94, wherein the oligosaccharide preparation comprises 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 anhydro subunit-containing oligosaccharides in relative abundance.

[0149] Item 96: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86-95, wherein each fraction of the oligosaccharide preparation comprises greater than 20%, 21%, 22%, 23%, 24%, or 25% of oligosaccharides containing anhydro subunits in relative abundance.

[0150] Item 97: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86-96, wherein greater than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35% or 30% of the anhydro subunit-containing oligosaccharides in the oligosaccharide preparation have only one anhydro subunit.

[0151] Item 98: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86-97, wherein the oligosaccharide preparation has a DP1 fraction content of 1% to 40% in relative abundance.

[0152] Item 99: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86-98, wherein the oligosaccharide preparation has a DP2 fraction content of 1% to 35% in relative abundance.

[0153] Item 100: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86-99, wherein the oligosaccharide preparation has a DP3 fraction content of 1% to 30% in relative abundance.

[0154] Item 101: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 100, wherein the oligosaccharide preparation has a DP4 fraction content of 0.1% to 20% in relative abundance.

[0155] Item 102: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 101, wherein the oligosaccharide preparation has a DP5 fraction content of 0.1% to 15% in relative abundance.

[0156] Item 103: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 102, wherein the ratio of the DP2 fraction to the DP1 fraction of the oligosaccharide preparation is 0.02-0.40 in relative abundance.

[0157] Item 104: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 103, wherein the ratio of the DP3 fraction to the DP2 fraction of the oligosaccharide preparation is 0.01-0.30 in relative abundance.

[0158] Item 105: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 104, wherein the aggregate content of the DP1 fraction and the DP2 fraction in the oligosaccharide preparation is less than 50%, 30% or 10% in relative abundance.

[0159] Item 106: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 105, wherein the oligosaccharide preparation comprises at least 103, 104, 105, 106 or 109 different oligosaccharide species.

[0160] Item 107: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 106, wherein two or more independent oligosaccharides of the oligosaccharide preparation comprise different anhydro subunits.

[0161] Item 108: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 107, wherein the oligosaccharide preparation comprises one or more anhydro subunits which are the product of reversible thermal dehydration of monosaccharides.

[0162] Item 109: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 108, wherein the oligosaccharide preparation comprises one or more dehydrated glucose, dehydrated galactose, dehydrated mannose, dehydrated allose, dehydrated altrose, dehydrated gulose, dehydrated idose, dehydrated talose, dehydrated fructose, dehydrated ribose, dehydrated arabinose, dehydrated rhamnose, dehydrated lyxose or dehydrated xylose subunits.

[0163] Item 110: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 109, wherein the oligosaccharide preparation comprises one or more anhydroglucose, anhydrogalactose, anhydromannose or anhydrofructose subunits.

[0164] Item 111: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 110, wherein the oligosaccharide preparation comprises one or more 1,6-anhydro-β-D-furanose or 1,6-anhydro-β-D-glucopyranose subunits. In some embodiments, the oligosaccharide preparation comprises both 1,6-anhydro-β-D-furanose and 1,6-anhydro-β-D-glucopyranose anhydro subunits.

[0165] Item 112: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 112, wherein the ratio of 1,6-anhydro-β-D-furanose to 1,6-anhydro-β-D-pyranose in the oligosaccharide preparation is about 10:1 to 1:10, 9:1 to 1:10, 8:1 to 1:10, 1 :4, 10:1 to 1 :3, 10:1 to 1 :2, or 1 :1 to 3:1.

[0166] Item 113: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 112, wherein the ratio of 1,6-anhydro-β-D-furanose to 1,6-anhydro-β-D-pyranose in the oligosaccharide preparation 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, 1:8, 1:9, or 1:10.

[0167] Item 114: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 113, wherein the ratio of 1,6-anhydro-β-D-furanose to 1,6-anhydro-β-D-pyranose in the oligosaccharide preparation is about 2:1.

[0168] Item 115: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 114, wherein the ratio of 1,6-anhydro-β-D-furanose to 1,6-anhydro-β-D-pyranose in each fraction of the oligosaccharide preparation is about 10:1 to 1:10, 9:1 to 1:10, 8:1 to 1:10, 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :10, 10:1 to 1 :11, 10:1 to 1 :12, 10:1 to 1 :13, 10:1 to 1 :2, or 1 :1 to 3:1.

[0169] Item 116: An oligosaccharide preparation for treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 115, wherein the ratio of 1,6-anhydro-β-D-furanose to 1,6-anhydro-β-D-pyranose in each fraction of the oligosaccharide preparation 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, 1:8, 1:9, or 1:10.

[0170] Item 117: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 116, wherein the ratio of 1,6-anhydro-β-D-furanose to 1,6-anhydro-β-D-pyranose in each fraction of the oligosaccharide preparation is about 2:1.

[0171] Item 118: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 117, wherein at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% of the anhydro subunits in the oligosaccharide preparation are selected from the group consisting of: 1,6-anhydro-β-D-furanose and 1,6-anhydro-β-D-pyranose.

[0172] Item 119: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 118, wherein the weight average molecular weight of the oligosaccharide preparation is about 300 g / mol to 5000 g / mol, 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 5000 g / mol, g / mol to 1500 g / mol, 300 g / mol to 2000 g / mol, 400 g / mol to 1300 g / mol, 400 g / mol to 1200 g / mol, 400 g / mol to 1100 g / mol, 500 g / mol to 1300 g / mol, 500 g / mol to 1200 g / mol, 500 g / mol to 1100 g / mol, 600 g / mol to 1300 g / mol, 600 g / mol to 1200 g / mol, or 600 g / mol to 1100 g / mol.

[0173] Item 120: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 119, wherein the number average molecular weight of the oligosaccharide preparation is about 300 g / mol to 5000 g / mol, 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 5000 g / mol, g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 2000 g / mol, 400 g / mol to 1000 g / mol, 400 g / mol to 900 g / mol, 400 g / mol to 800 g / mol, 500 g / mol to 900 g / mol, or 500 g / mol to 800 g / mol.

[0174] Item 121: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 120, wherein the degree of polymerization distribution is determined and / or detected by MALDI-MS, GC-MS, LC-MS, SEC, HPLC and / or a combination thereof (e.g., MALDI-MS and SEC).

[0175] Item 122: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 121, wherein the degree of polymerization of the oligosaccharide preparation can be determined based on its molecular weight and molecular weight distribution.

[0176] Item 123: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 122, wherein the oligosaccharide preparation is contained in a nutritional composition at a concentration of at least 50 g / ton of feed (e.g. at least 70 g / ton of feed, 100 g / ton of feed, 200 g / ton of feed, 300 g / ton of feed, 400 g / ton of feed, 500 g / ton of feed, 600 g / ton of feed, 700 g / ton of feed, 800 g / ton of feed, 900 g / ton of feed) and / or wherein the oligosaccharide preparation is contained in the nutritional composition at an inclusion rate of at least 50 ppm (e.g., at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 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, 500 ppm).

[0177] Item 124: An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis according to any one of items 86 to 123, wherein the oligosaccharide preparation is administered for at least one day, preferably at least 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 days 4 days, 45 days, 46 days, 47 days, 48 ​​days, 49 days, 50 days, 51 days, 52 days, 53 days, 54 days, 55 days, 56 days, 57 days, 58 days, 59 days, 60 days, 61 days, 62 days, 63 days, 64 days, 65 days, 66 days, 67 days, 68 days, 69 days, 70 days, 71 days, 72 days, 73 days, 74 days, 75 days 5 days, 76 days, 77 days, 78 days, 79 days, 80 days, 81 days, 82 days, 83 days, 84 days, 85 days, 86 days, 87 days, 88 days, 89 days, 90 days, 91 days, 92 days, 93 days, 94 days, 95 days, 96 days, 97 days, 98 days, 99 days, 100 days, 101 days, 102 days, 103 days, 104 days, 105 days 05 days, 106 days, 107 days, 108 days, 109 days, 110 days, 111 days, 112 days, 113 days, 114 days, 115 days, 116 days, 117 days, 118 days, 119 days, 120 days, 121 days, 122 days, 123 days, 124 days, 125 days, 126 days, 127 days, 128 days, 129 days, 130 days, 131 days, 132 days, 133 days, 134 days, 135 days, 136 days, 137 days, 138 days, 139 days, 140 days, 141 days, 142 days, 143 days, 144 days, 145 days, 146 days, 147 days, 148 days, 149 days, 150 days, 151 days, 152 days, 153 days, 154 days, 155 days, 156 days, 157 days, 158 days, 159 days, 160 days, 161 days, 162 days, 163 days, 164 days, 165 days, 166 days, 167 days, 168 days, 169 days, 170 days, 171 days, 172 days, 173 days, 174 days, 175 days, 176 days, 177 days, 178 days, 179 days, 180 days, 181 days, 182 days, 183 days, 184 days, 185 days, 186 days, 187 days, 188 days, 189 days, 190 days, 191 days, 192 days, 193 days, 194 days, 195 days, 196 days, 197 days, 198 days, 199 days, 200 days, 201 days, 202 days, 203 days, 204 days,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 the nutritional composition is administered continuously (i.e., uninterruptedly).

[0178] The characteristics of the oligosaccharide preparations mentioned herein may be any one, two or more or even all of the characteristics of the individual oligosaccharide preparations as described in the above items. In other words, the characteristics of the oligosaccharide preparations may be any combination of the individual characteristics of the oligosaccharide preparations described in the above items. For example, in a specific embodiment of the method or use according to the present invention, the characteristics of the oligosaccharide preparations may be a combination of the characteristics of the combined oligosaccharide preparations as described in items 9, 11, 20 and 39.

[0179] In the following, the present invention is further described by non-limiting examples.The present invention as disclosed herein is not limited to the specific embodiments, methodologies, examples, schemes, etc. described herein, but is only limited by the claims. Example

[0180] Example 1 - Effects of oligosaccharide preparations on birds challenged with necrotic enteritis

[0181] Feeding trials were performed to investigate the effects of oligosaccharide preparations on zootechnical and biological parameters of broiler chickens subjected to a challenge model of necrotic enteritis produced by inoculation with Eimeria and Clostridium perfringens.

[0182] The oligosaccharide preparation is prepared as disclosed in WO 2020 / 097458 and WO 2016 / 007778, in particular in any one of Examples 1 to 7, 16 to 18 of WO 2020 / 097458A1, in the method described in paragraph

[317] , and / or in any one of Examples 73 to 77, 80 to 89, 97 to 99, 101 to 110 of WO 2016 / 007778 A1. The oligosaccharide preparation comprises at least n oligosaccharide fractions, each oligosaccharide fraction having a different degree of polymerization selected from 1 to n (DP1 fraction to DPn fraction), wherein n is an integer greater than 3; wherein each of the DP1 fraction and the DP2 fraction independently comprises about 0.5% to about 15% of anhydro subunit-containing oligosaccharides in relative abundance as determined by mass spectrometry. Furthermore, the oligosaccharide preparations produced may also be characterized according to the characteristics described herein.

[0183] In the feeding trial, 1600 birds were allocated to floor pens with 25 birds per pen. Treatments were replicated in 8 blocks with 8 pens randomly allocated within each block. Birds were allocated to pens to achieve equal starting pen weights for each treatment. One pen was considered one experimental unit.

[0184] The duration of the trial was 42 days. All birds were vaccinated against coccidiosis (Merck Animal Health USA -B52, a live coccidial vaccine isolated from chickens and prepared from coccidia-susceptible strains of E. acervulina, E. maxima, E. maxima MFP, E. mivati, and E. tenella according to the product bulletin).

[0185] The negative control group (NC) was not challenged with necrotic enteritis and was not administered with an oligosaccharide preparation. The challenge control group (CC) was challenged with necrotic enteritis but was not administered with an oligosaccharide preparation. The first test group (OP500) was challenged with necrotic enteritis and fed a feed containing 500 ppm of an oligosaccharide preparation. The second test group (OP700) was challenged with necrotic enteritis and fed a feed containing 700 ppm of an oligosaccharide preparation. The third test group (OP900) was challenged with necrotic enteritis and fed a feed containing 900 ppm of an oligosaccharide preparation.

[0186] Animal Management : The statistical characteristics of the birds were as follows: Genetic: Cobb 500; Sex: Male; Starting age: Day of hatching; Only healthy-looking birds were used in this study.

[0187] The temperature of the building was monitored. Environmental conditions (temperature) during the experiment were appropriate (optimal) for the age of the animals. Lighting was provided by fluorescent lamps above the pens. The lighting scheme was 22 hours of light and 2 hours of darkness per day. Standard ground pen management practices were used throughout the experimental period. Animals and housing facilities were inspected twice daily to observe and record general health, constant supply of feed and water, and temperature, removal of all dead birds, and identification of emergencies.

[0188] Check pens daily for dead birds. Kill birds only to relieve suffering. When birds are killed or found dead, record the pen, date, and weight (kg) of the kill. Perform autopsies on all dead or killed birds to determine the probable cause of death.

[0189] Feed and Water: The number of test articles produced in each batch was recorded. Feeds were mixed to ensure uniform distribution of the test articles. Experimental diets were formulated from a uniform basal diet. All treatment feeds were mixed using a Davis S-20 mixer and pelleted using a California Pellet Mill. Broiler diets were fed in the form of crumbles (starter feed) and pellets (grower and finisher feed). Diet formulations are included in the raw data. Feed allotment = 5 kg / bird x 200 birds / batch = 1,000 kg. Diets were provided ad libitum in one tube feeder per pen. From day 1 to day 7, feed was also provided in a feeding pan placed on the litter in each pen. Water was provided ad libitum. All feeds were weighed per pen. Starter feed was fed from day 0 to day 21. On day 14, unconsumed starter feed was weighed and re-issued. On day 21, unconsumed starter feed was weighed and discarded. Grower feed was issued and fed until day 35. On day 35, unconsumed grower feed was weighed and discarded. Finisher feed was distributed and fed until day 42. The diet formula is shown in Table 1 below.

[0190] Table 1: Diet formulation

[0191]

[0192]

[0193] Test Procedure Three feed samples (approximately 2 lbs each) were collected: one from the beginning, middle, and end of each treatment diet batch and then mixed to form a composite sample (2 lbs). The composite sample was divided into 3 equal aliquots. After the study was completed, one composite sample of each treatment diet was retained for 30 days and the other two were used for analysis of diet composition.

[0194] Necrotic enteritis challenge model (expected NE mortality = 5-10%): All pens except NC are challenged with necrotic enteritis, e.g., Eimera and C. perfringens. On day 14, each bird is injected with 5,000 oocysts of Eimeria maxima. This isolate is from the wild type and has been identified to provide weak aggressiveness. The oocyst inoculum is mixed into the feed at the bottom of the tube feeder. On days 19, 20, and 21, all birds are injected with a broth culture of Clostridium perfringens (CP6). This wild type isolate is from a commercial broiler farm. Fresh inoculum is used daily, with a titer level of approximately 1.0 x 10 8-9 / head. Inoculum was added by mixing into the feed at the bottom of the tube feeder. The number of days CP was used depended on the mortality observed each day. Three days of dosing were used.

[0195] Coccidian Oocysts per Gram of Litter (OPG): Baseline litter samples were collected on Day 0 to determine baseline OPG. Fresh fecal samples were collected from each pen on Days 14, 21, 28, and 35. These representative samples were tested to determine the extent of oocyst shedding / circulation. The number of oocysts per gram of sample was determined.

[0196] Necropsy Sample Collection: On day 21, two birds per pen were randomly selected, sacrificed, weighed individually, and examined for the presence of necrotic enteritis (NE) lesions. Ileal sections were collected from each bird in formalin. The formalin was exchanged for ethanol after 24 hours, and these fixed samples were subjected to histological analysis as determined necessary after analysis of performance data. Similarly, cecal samples were collected and stored at -80°C and subsequently analyzed based on performance data.

[0197] Pen weights and feed disappearance: Pen weights and feed disappearance were recorded on days 14, 21, 35, and 42. Feed conversion ratio was calculated.

[0198] Mortality: Birds found dead during the study were recorded and not replaced. Pen number, date of death, sex, weight, and diagnosis were recorded. No birds were replaced during the study.

[0199] Data analysis : Outcome variables were i) performance (pen and feeder weights on days 14, 21, 35 and 42. Cumulative BWG and FCR were calculated); ii) OPGs (oocysts per gram of feces); iii) mortality (number of birds confirmed to be NE-related and total mortality); iv) lesion scores.

[0200] Data and analysis: Mean values ​​of live weight, weight gain, feed consumption, feed conversion ratio (FCR), OPGs, and mortality (total and NE mortality) were calculated. Data were analyzed by ANOVA using JMP data analysis software, and means were differentiated using Tukey's test (P < 0.05).

[0201] result : Supplementation with oligosaccharide preparations increased body weight gain (BWG) and feed conversion ratio (FCR) of birds compared to the challenged control group (CC) over the entire experimental period from 0 to 42 days of age. The necrotic enteritis challenged groups supplemented with oligosaccharide preparations (OP500, OP, 700, OP900) were statistically similar to the non-challenged control group NC. The CC group had a higher mortality rate associated with necrotic enteritis (NE) compared to the NC group. Administration of oligosaccharide preparations reduced mortality associated with necrotic enteritis compared to the CC group. The CC group had a higher score of NE characteristic lesions than any other group. Notably, the lesion scores were reduced in the groups administered with oligosaccharide preparations compared to the CC group. In particular, in the necrotic enteritis challenge, the CC group had a higher frequency of grade 1 and 2 lesions compared to the NC group, while the frequency of grade 0 lesions was significantly reduced. However, the groups receiving oligosaccharide preparations (OP500, OP700, OP900) reduced the number of birds with grade 1 and 2 lesions and increased the number of birds with grade 0 lesions. Thus, administration of oligosaccharide preparations was found to reduce the severity of lesions under NE challenge. Table 2 summarizes the results.

[0202] In conclusion, the addition of oligosaccharide preparations in all groups was beneficial in reducing the negative effects of necrotic enteritis in birds. It was even found that the administration of 900 ppm of the oligosaccharide preparations completely restored the growth performance of challenged birds and reduced NE-related mortality similar to that of non-challenged birds.

[0203] Table 2: Body weight gain (BWG), feed intake (FI), feed conversion ratio (FCR), NE-related mortality (NE-mortality), foot pad lesions (FPD) and lesion scores of broiler chickens.

[0204]

Claims

1. A method for reducing the effects of necrotic enteritis in poultry, comprising the step of administering an oligosaccharide preparation to poultry in need of reducing the effects of necrotic enteritis, wherein the oligosaccharide preparation comprises at least n oligosaccharide fractions, each oligosaccharide fraction having a different degree of polymerization selected from 1 to n (DP1 fraction to DPn fraction), wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90% (e.g., 1% to 90%; or such as about 0.5% to about 15%) anhydro subunit-containing oligosaccharides in relative abundance as determined by mass spectrometry.

2. The method of claim 1, wherein the effect of necrotic enteritis is at least one selected from the group consisting of: reduced weight gain in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds not requiring reduction of the effect of necrotic enteritis; increased feed conversion rate in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds; increased mortality in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds; and increased lesion scores in birds requiring reduction of the effect of necrotic enteritis compared to a control group of birds.

3. A method according to claim 1 or 2, wherein the poultry in which the effects of necrotic enteritis are to be reduced are challenged with Clostridium perfringens.

4. A method according to any preceding claim, wherein the oligosaccharide preparation is comprised in a nutritional composition for administration to poultry.

5. The method according to any one of the preceding claims, wherein the oligosaccharide preparation is included in the nutritional composition at an inclusion rate of at least 50 ppm (e.g. at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm).

6. A method according to any one of the preceding claims, wherein the oligosaccharide preparation is administered to birds before, after and / or simultaneously with a diet of the birds in need of reducing the effects of necrotic enteritis, preferably simultaneously with the diet.

7. The method according to any one of the preceding claims, wherein the bird is poultry, such as chicken, duck, goose, turkey, guinea fowl, pigeon, quail; preferably broiler chickens.

8. The method of any one of the preceding claims, wherein the effects of necrotic enteritis are reduced on at least one day within 42 days after the first administration of the oligosaccharide preparation (on one or more days selected from the group consisting of 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days after administration of the oligosaccharide preparation).

9. The method of any one of the preceding claims, wherein n of the oligosaccharide preparation 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.

10. The method of any of the preceding claims, wherein at least one fraction of the oligosaccharide preparation 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 anhydro subunit-containing residues in relative abundance. and / or wherein each fraction of the oligosaccharide preparation 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 anhydro subunits in relative abundance.

11. The method of any one of the preceding claims, wherein the weight average molecular weight of the oligosaccharide preparation is about 300 g / mol to 5000 g / mol (e.g., about 2000 g / mol to 2800 g / mol, 2100 g / mol to 2700 g / mol, 2200 g / mol to 2600 g / mol, 2300 g / mol to 2500 g / mol, or 2320 g / mol to 2420 g / mol), 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 2000 g / mol, 400 g / mol to 5000 g / mol g / mol, 1100 g / mol, 1900 g / mol, 2000 g / mol, 2100 g / mol, 2200 g / mol, 2400 g / mol, 2500 g / mol, 2600 g / mol, 2700 g / mol, 2800 g / mol, 2900 g / mol, 3000 g / mol, 3100 g / mol, 3200 g / mol, 3300 g / mol, 3400 g / mol, 3600 g / mol, 3700 g / mol, 3800 g / mol, 3900 g / mol, 400 g / mol, 4100 g / mol, 4200 g / mol, 4300 g / mol, 4400 g / mol, 450 g / mol, 4600 g / mol, 4700 g / mol, 4800 g / mol, 4900 g / mol, 500 g / mol, 5100 g / mol, 5100 g / mol, 5200 g / mol, 5200 g / mol 12. The method according to any one of the preceding claims, wherein the relative abundance of oligosaccharides in each of the n fractions of the oligosaccharide preparation decreases monotonically with its degree of polymerisation.

13. Use of an oligosaccharide preparation to reduce the effects of necrotic enteritis in poultry in need thereof, wherein the oligosaccharide preparation comprises at least n oligosaccharide fractions, each oligosaccharide fraction having a different degree of polymerization selected from 1 to n (DP1 fraction to DPn fraction), wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90% (e.g., 1% to 90%; or such as about 0.5% to about 15%) anhydro subunit-containing oligosaccharides in relative abundance as determined by mass spectrometry.

14. An oligosaccharide preparation for use in treating, ameliorating and / or preventing poultry suffering from and / or at risk of necrotic enteritis, wherein the oligosaccharide preparation comprises at least n oligosaccharide fractions, each oligosaccharide fraction having a different degree of polymerization selected from 1 to n (DP1 fraction to DPn fraction), wherein n is an integer greater than or equal to 2; and wherein each fraction comprises at least about 0.5% to about 90% (e.g., 1% to 90%; or such as about 0.5% to about 15%) anhydro subunit-containing oligosaccharides in relative abundance as determined by mass spectrometry.

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