Application of human milk oligosaccharide in calf fattening

Abstract

The invention provides an application of human milk oligosaccharide in calf fattening. The present invention relates to the use of human milk oligosaccharides for feeding production animals such as calves. The invention also relates to a milk replacer composition containing human milk oligosaccharides. Another subject of the invention is a method of preventing and treating gastrointestinal diseases, in particular gastrointestinal infections, in young production animals, in particular calves, by administering an effective amount of one or more human milk oligosaccharides. In the context of the present invention, 2 '-fucosyllactose (2-FL) is particularly useful as one or said human milk oligosaccharide.

Description

[0001] This application is a divisional application of Chinese Patent Application No. 201780052697.9, entitled "Use of human milk oligosaccharides in calf fattening", filed on August 29, 2017. Technical Field

[0002] This invention relates to the use of human milk oligosaccharides (HMIs) in feeding production animals such as calves. The invention also relates to milk substitute compositions comprising HMIs. Another subject of the invention is a method for preventing and treating gastrointestinal diseases (particularly gastrointestinal infections) in young production animals (particularly calves) by administering an effective amount of one or more HMIs. In the context of this invention, 2'-fucosylated lactose (2-FL) is particularly useful as one or more of the aforementioned HMIs. Background Technology

[0003] The feeding of healthy calves is the foundation of success in the dairy industry. Achieving healthy calves through optimal agriculture and care, early development of gastroesophageal vestibular function, and economically sound feeding are several goals of calf rearing (Steinwunder et al. (2005) Milchviehfütterung, Tier- und Leistungsgerecht. Leopold StockerVerlag, Graz; Drochner, W. (2008): Fütterung der Rinder. in: Jeroch et al. (ed.) Ernahrung landwirtschaftlicher Nutztiere, 2nd ed. Eugen Ulmer KG, Stuttgart, pp. 385-467).

[0004] Successful feeding is associated with minimizing animal losses, which is considered the most critical issue. In Germany, losses have increased by 8% to 14% over the years (see, for example, Röhrmoser (1995): Aufzucht des Rindes. in: Matzke et al. (eds.) Wirtschaftliche Milchviehhaltung und Rindermast. 3rd ed., Frankfurt am Main, Verlag Union Agrar, pp. 221-261). The target should be to achieve losses of approximately 5%. Optimized calf feeding can help achieve this goal. Several preventative measures have been proposed in feeding, such as reducing the incidence of diarrhea in calves (Schrag et al. (1984) Gesunde Kälber-Gesunde Rinder. Die wichtigsten Krankheiten in Aufzucht und Mast. Erkennung, Vorbeuge, Behandlung. 3rd ed. Henbersberg, Schober Verlags-GmbH, pp. 249-276). These measures focus on a well-developed gut microbiota and gut health. The inclusion of various prebiotics in milk replacers has been proposed to improve and maintain gut health (Dohms (2004) Aspekte der Darmgesundheit und Chancen fur den Einsatz von Probiotika, 4th ed.).

[0005] Human milk oligosaccharides (HMOs) are a rich and unique family of unbound glycans in human milk; for example, in the following review: Bode (2012) Glycobiology 22 (9), 1147-1162.

[0006] WO-A-2012 / 158517 discloses the use of human milk oligosaccharides, particularly 2'-fucosylated lactose, for promoting the growth of bacteria found in infant feces. This prior art also intends to use human milk oligosaccharides as prebiotics for the nutrition of various mammals; however, no experimental data are provided beyond the promotion of Enterobacter growth, therefore the use of human milk oligosaccharides to feed animals remains speculative. Summary of the Invention

[0007] The technical problem to be solved by the present invention is to provide improved feeding of young producing animals, especially calves, particularly by improving their feeding.

[0008] The solutions to the above-mentioned technical problems are provided by the embodiments of the present invention described herein, and are as defined in the claims.

[0009] In particular, the present invention relates to novel milk substitute compositions comprising at least one human milk oligosaccharide. Detailed Implementation

[0010] According to the present invention, a "human milk oligosaccharide" is a sugar molecule composed of at least three carbohydrate entities, which is present in female breast milk or pre-milk. Preferably, an HMO is an oligosaccharide composed of 3, 4, 5, 6, 7, 8, 9, 10 or more sugar entities, and oligosaccharides containing more than 4 entities can be linear or branched oligomers. HMOs are typically composed of monosaccharides glucose, galactose, N-acetylglucosamine, N-acetyllactosamine, fucose, and N-acetylneuraminic acid. Most HMOs used in the present invention are based on the disaccharide lactose or N-acetyllactosamine, and lactose-based HMOs are particularly preferred.

[0011] In the context of this invention, preferred human milk oligosaccharides include, but are not limited to, 2'-fucosyllactose (2-FL), 3'-fucosyllactose (3-FL), 3'-sialyllactose (3-SL), 6'-sialyllactose (6-SL), lact-N-tetrasaccharide (LNT), lact-N-neotetrasaccharide (LNnT), lact-N-hexasaccharide (LNH), isoylacto-N-octasaccharide, isoylacto-N-neooctasaccharide, p-lacto-N-octasaccharide, lact-N-fucopentose I (LNFP I), lact-N-fucopentose II (LNFP II), lact-N-fucopentose III (LNFP III), lact-N-fucopentose V (LNFP V), LS-tetrasaccharide a (LST a), LS-tetrasaccharide b (LST b), LS-tetrasaccharide c (LST c), and disialyllacto-N-tetrasaccharide (DSLNT).

[0012] Particularly preferred HMOs for use in this invention are human lactotrisaccharides 3-FL and 2-FL. Recently, biotechnological production of 2-FL has been provided (see WO-A-2010 / 070104), making this basic HMO available on an industrial scale (Jennewein Biotechnologie GmbH, Rheinbreitbach, Germany). Therefore, the HMOs in the milk substitute composition are preferably synthetic HMOs prepared in bacterial cultures, particularly using genetically modified bacteria taught in WO-A-2010 / 070104. In the context of this invention, 2-FL, particularly synthetic 2-FL as described above, and higher HMOs based on 2-FL are generally preferred, and 2-FL is the most preferred HMO for carrying out this invention.

[0013] This invention also relates to milk substitute compositions comprising more than one specific HMO, wherein a mixture of two or more of the aforementioned HMOs is preferred. Particularly preferred are HMO mixtures in which one of the HMOs is 2-FL or 3-FL, or the mixture comprises 2-FL and 3-FL, as well as a third, fourth, fifth, or other HMO. In the context of this invention, a combination of 2-FL and 3-FL is most preferred.

[0014] Of course, the milk substitute compositions of the present invention may also contain sugars other than HMOs. Lactose and N-acetyllactosamine are mentioned herein as preferred other oligosaccharides.

[0015] "Milk replacer" or "milk replacer composition" refers to a nutritional composition for feeding production animals, preferably selected from cattle, goats, sheep, pigs, deer and horses, especially infants and young production animals. According to the invention, the terms "milk replacer" and "milk replacer composition" also include the definition provided in Article 3(2) of Regulation (EC) 767 / 2009, wherein "milk replacer" means "a compound feed, which is administered in dry form or diluted in a given amount of liquid, as a supplement or substitute for postmilk for feeding young animals, or for feeding young animals such as calves, lambs or goats for slaughter."

[0016] Therefore, the milk replacer compositions of the present invention typically comprise one or more HMOs as described above, as well as a base milk replacer composition. The term "milk replacer composition" in the present invention refers to compositions in both dry and liquid forms, the liquid form typically obtained by reconstituted the dry composition with water. The base nutritional compositions used to provide the milk replacer compositions of the present invention are known in the art (for an overview of calf milk replacers, see, for example, Bovine Alliance on Management and Nutrition (BAMN), 2008, A GUIDE TO CALF MILK REPLACERS, available online at https: / / www.aphis.usda.gov / animal_health / nahms / dairy / downloads / bamn / BAMN08_GuideMilkRepl.pdf; and Kamphues et al. (2004) Supplemente zu Vorlesungen und Ubungenin der Tierernahrung, Hannover, Verlag M. & H. Schaper Alfred).

[0017] Therefore, the milk substitute compositions of the present invention generally contain at least protein and fat sources, which are optionally reconstituted in an aqueous liquid (wherein the components may be present in the form of suspensions, emulsions and / or solutions in an aqueous base), and are generally mixed with other ingredients such as carbohydrates, fiber, vitamins, mineral supplements and other additives such as pharmaceuticals, emulsifiers, thickeners, trace elements, antioxidants and flavorings.

[0018] Milk replacer compositions are typically classified according to the source of their protein and / or fat components. Typical protein sources for milk replacer compositions, particularly those intended for calf feeding, include skim milk powder, whey powder, and plant protein sources such as soy protein, soy flour, wheat gluten, or wheat isolate. The fat component of milk replacers can be derived from animal fats or vegetable oils.

[0019] Protein and fat levels are important characteristics of the milk replacer compositions used in this invention. Preferred protein levels in calf milk replacers are typically about 18% by weight to about 22% by weight, most preferably about 20% by weight, and preferred fat levels are about 10% by weight to about 28% by weight, more preferably about 18% by weight to about 23% by weight. The levels of these components also vary depending on the type of animal being fed. For example, for breeding calves, the fat level is typically about 13% by weight to about 20% by weight, while for fattening calves (i.e., calves intended for slaughter), the fat level in the milk replacer composition should be increased to about 23% by weight. A special consideration in producing the milk replacer composition is the carbohydrate-to-protein ratio. For example, pure dried whey contains about 70% by weight of lactose and about 12% by weight of protein. To implement this invention, the carbohydrate content, particularly lactose, glucose, and sucrose (excluding HMO content), should preferably not exceed about 50% by weight to prevent undue fermentation in the colon due to high throughput (Kamphues (2004), ibid.).

[0020] A typical basic dairy substitute composition used in this invention comprises about 22% by weight or more crude protein, about 20% by weight or more crude fat, no more than about 0.15% by weight crude fiber, about 0.75% by weight to about 1.75% by weight calcium, at least about 0.7% by weight phosphorus, at least about 20,000 IU / Ib of vitamin A, at least about 5,000 IU / Ib of vitamin D3, and about 100 IU / Ib of vitamin E.

[0021] Based on the total weight of the composition, the milk substitute composition according to the present invention generally contains about 0.5% to 10.0% by weight, preferably about 1% to 5% by weight, and most preferably 4% by weight of HMO (which may be a single type of HMO, such as 2-FL, or a mixture of two or more different HMOs).

[0022] This invention also relates to the use of HMOs (one or more HMOs) for feeding production animals, particularly young production animals such as calves, lambs, goats, and foals. HMOs are particularly useful in calf fattening.

[0023] This invention also relates to a method for feeding production animals, particularly young production animals such as calves, lambs, goats, and foals, comprising the step of feeding the production animals with the milk replacer composition of this invention. The feeding method of this invention improves feed intake and performance parameters of production animals, particularly in the case of calf fattening. A particular benefit of using HMOs as an ingredient in the milk replacer compositions taught herein is the improvement in the overall condition of the fed production animals, particularly calves, lambs, goats, and foals, thereby improving the performance parameters of animals fed with the milk replacer according to this invention. The milk replacer compositions of this invention reduce animal losses, particularly by preventing intestinal diseases that cause diarrhea, such as bacterial or viral intestinal infections. Without wishing to be bound by any theory, it is believed that HMOs improve the growth of beneficial gut bacteria, thereby reducing the relative number of pathogens, while preventing or at least hindering the binding of pathogens such as pathogenic Enterobacteriaceae and viruses to intestinal receptors. HMOs are also believed to improve the gut microbiota of animals, modulate mucosal immunity, and enhance the animal's resistance to atopic conditions.

[0024] In the context of this invention, HMOs are particularly useful in improving the health of production animals such as calves, lambs, goats, and foals. In particular, HMOs (which, as described above, can be administered alone or as a mixture of HMOs) are used to prevent and treat, and at least reduce, the occurrence (or at least shorten the duration) of gastrointestinal diseases, especially bacterial and viral gastrointestinal diseases (such as diarrhea), in said production animals. HMOs are also useful in preventing and treating atopic diseases in production animals (preferably those defined above), which are conditions associated with a hypersensitive response of the body to foreign substances. Furthermore, HMOs are useful in improving mucosal immunity in production animals (particularly those mentioned above). According to the invention, HMOs also improve the gut microbiota of production animals such as calves, lambs, goats, and foals. HMOs can be administered alone or in the form of a composition, preferably orally, and can also be in the form of a milk replacer composition as defined herein. Pharmaceutical compositions can also be in the form of tablets or capsules administered separately from any feeding composition. Therefore, the present invention also relates to a method for preventing or treating gastrointestinal diseases (e.g., bacterial and viral infections of the gastrointestinal system, such as diarrhea) in producing animals (preferably juvenile producing animals as described above), the method comprising the step of administering to the producing animal a safe and effective amount of at least one HMO or a composition containing two or more HMOs. The present invention also relates to the use of at least one HMO or a composition containing two or more HMOs in the preparation of medicaments for the prevention or treatment of the aforementioned conditions.

[0025] The invention is further illustrated by the following non-limiting embodiments: Example Example 1: Feeding breeding calves with milk replacer containing HMOs Materials and Methods: Sixty calves (Deutsche Holstein) were divided into two groups (control group and experimental group, 30 calves per group), with equal numbers of males (15 / 30) and females (15 / 30). The mean age of the calves was 7 days. Each animal in both groups was kept individually fed. At 14 days of age, the calves were divided into two groups, each kept in a different but otherwise identical barn. The milk replacer composition was fed to the animals via an automatic waterer. The animals had free access to water from the waterer. In addition, each animal was given 150 g of hay per day. From week 8, a concentrated feed was given via an automatic dispenser. Both groups were fed the same milk replacer composition except for HMO. The experimental group was fed the basal milk replacer composition plus 4 wt% 2-FL. The control group was fed an additional 4 wt% whey powder instead of HMO. One liter of liquid milk replacer composition contained 125 g of dry milk replacer concentrate.

[0026] The following table shows the feeding protocol:

[0027] Experiment duration: 77 days

[0028] Data collection: Feeding via an automated feeder with individual animal identification, measuring food intake for each individual; collecting hay and feed concentrate data by weighing; weighing each animal daily; measuring body temperature under the tongue at the automated feeder and rectal measurement weekly for comparison and control; controlling health status and medication use; weekly analysis of feces for intestinal bacterial colonization.

[0029] Feed analysis. Analysis of milk replacers, hay, and feed concentrates.

[0030] Example 2: Feeding fattening calves with milk replacer containing HMOs

[0031] Materials and Methods: Sixty breeding Fleckvieh calves were divided into two groups (control / experiment), with 30 calves in each group. Both groups contained equal numbers of male (15 / 30) and female (15 / 30) calves. The average age of the animals was 14 days (live weight 50-60 kg). Each group of animals was kept in an identical barn. The animals were fed a milk replacer composition via an automatic water dispenser. The animals had free access to water. In addition, each animal was given 300 g of hay daily. Both groups were fed the same milk replacer composition except for HMO. The experimental group was fed a basal milk replacer composition plus 4% wt% 2-FL. The control group was fed an additional 4% wt% whey powder instead of HMO. At the start of the experiment, 1 liter of liquid feed composition contained 125 g of dry milk replacer concentrate. At the end of the experiment, the concentration of the liquid feed composition was increased to 250 g / L.

[0032] The feeding plan is as follows:

[0033] Experiment duration: The experiment began at approximately 14 days of age and ended with a final slaughter weight of approximately 230 kg to 240 kg (around the 22nd week of the experiment), lasting approximately 140 days.

[0034] Data collection: Feeding is conducted via an automated feeder with individual animal identification, measuring each individual's food intake; hay and feed concentrate data are collected by weighing; each animal is weighed daily; body temperature is measured sublingually at the automated feeder and rectally once a week for comparison and control; health status and medication use are monitored; fecal samples are analyzed weekly for intestinal bacterial colonization. Post-slaughter, each animal's physical condition is assessed.

[0035] Feed analysis. Analysis of milk replacer and hay.

Claims

1. Use of a milk substitute composition containing 2'-fucosylated lactose in the feeding of production animals, wherein the 2'-fucosylated lactose is prepared in bacterial culture; wherein the content of carbohydrates other than human milk oligosaccharides in the composition does not exceed 50% by weight.

2. The use as described in claim 1, wherein, The production animal is any one of calves, lambs, goats, and foals.

3. The use as described in claim 1, wherein, The animal used for production is a calf.

4. The use as described in claim 1, wherein, The production animal is a fattening calf.

5. The use as described in claim 4, wherein, The dairy substitute composition containing 2'-fucosylated lactose is used to regulate mucosal immunity in fattening calves.

6. The use as described in claim 1, wherein, The composition has a protein level of 18% to 22% by weight and a fat level of 10% to 28% by weight.

7. Use of a milk substitute composition containing 2'-fucosylated lactose in the preparation of a medicament for treating or preventing gastrointestinal diseases in production animals, wherein the 2'-fucosylated lactose is prepared in a bacterial culture; and wherein the composition contains no more than 50% by weight of carbohydrates other than human milk oligosaccharides.

8. The use as described in claim 7, wherein, The production animal is any one of calves, lambs, goats, and foals.

9. The use as described in claim 7, wherein, The production animal is a fattening calf.

10. The use as described in claim 9, wherein, The dairy substitute composition containing 2'-fucosylated lactose is used to regulate mucosal immunity in fattening calves.

11. The use as described in claim 7, wherein, The composition has a protein level of 18% to 22% by weight and a fat level of 10% to 28% by weight.

12. A method of feeding fattening calves, comprising the step of feeding the fattening calves a milk substitute composition containing 2'-fucosylated lactose, wherein the 2'-fucosylated lactose is prepared in a bacterial culture; wherein the content of carbohydrates other than human milk oligosaccharides in the composition does not exceed 50% by weight.

13. The method of claim 12, wherein, The composition has a protein level of 18% to 22% by weight and a fat level of 10% to 28% by weight.

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