Ruminant feed rich in particles of rumen labile components

Abstract

The present invention relates to a ruminant feed rich in particles of rumen labile components. The present invention relates to a granular ruminant feed in the form of granules or subsequently comminuted form, comprising a mixture of at least one solid granulated feed component and at least one rumen-labile component (added to the mixture). The invention also relates to a method for preparing the ruminant feed of said granules and to the use thereof in a method for modifying the milk fat concentration.

Description

[0001] This application is a divisional application of PCT application PCT / EP2015 / 055006, filed on March 11, 2015, with the invention title of "Ruminant Feed Containing Rumen-Unstable Components in Granular Form", and the date of entry into the Chinese national phase of the PCT application is September 12, 2016, with the application number 201580013314.8.

[0002] The present invention relates to a ruminant feed in granular form, which is a mixture of at least one solid particulate feed component and at least one rumen-unstable component (which has been added to the mixture); the present invention also relates to a method for preparing the same and its use in a method for altering the milk fat concentration. Background of the Invention

[0004] Ruminant feed additives may be subject to undesired chemical modification by ruminant organisms. For example, in the context of rumen microbial modification of polyunsaturated fatty acids, it can be observed that the content and position of double bonds on the fatty acid carbon chain may change. Generally, the double bonds of unsaturated fatty acids in plants are separated by at least two single bonds. Microbial enzymes are capable of generating a conjugated double bond system, in which the double bonds are separated by only one single bond. They include, for example, conjugated linoleic acid isomers (conjugated linoleic acid, CLA). They are mainly formed from linoleic acid (C18:2 cis-9, trans-12) by the linoleic acid isomerase of the rumen bacterium Butyrivibrio fibrisolvens. The main CLA isomer is cis-9, trans-11-CLA (C18:2 cis-9, trans-11), which accounts for about 75-92% of the total CLA content. In contrast, the milk fat-lowering isomer C18:2 trans-10, cis-12 is present only in small amounts of 0.03-1.5%. Some of these isomers are absorbed into the tissues of ruminants. In the case of dairy cows, rumen-modified fatty acids are also transported into the milk. The CLA content of milk fat is highly feed-dependent and accounts for about 0.3-1.1% (see: Kirchgeβner, 13th Edition, DLG Verlag GmbH), and even exceeds 4% in animals fed on pasture ( et al., 2014; in press).

[0005] Due to their effective physiological or pharmacological activities, CLAs have been intensively studied. CLAs have shown cancer defense and anti-inflammatory activities in cell culture studies and animal tests. It is also known that conjugated linoleic acid isomers (CLA isomers) inhibit the expression of a large number of genes in ruminants, which are responsible for the synthesis of fatty acids and the intake of cyclic fatty acids that form triglycerides into the mammary gland. In particular, it has been confirmed in the context that the C18:2 trans-10, cis-12 CLA isomer particularly reduces fat. The advantage of the discussion related to reducing milk fat content lies particularly in slowing down metabolism and increasing milk production. It has also been reported that a 0.4 - 0.5% reduction in milk fat content can lead to an increase in dietary production of up to 3 - 10% (see Kirchgeβner, see above).

[0006] This is why CLAs are currently commonly added to milk-producing feed (Milchleistungsfutter). Although these products have been approved for feeding, they must be in a rumen-protected form because unprotected CLAs are further degraded in the rumen into inactive C18:1 and C18:0 fatty acids.

[0007] When these or other molecules that are degraded by rumen microorganisms or chemically modified and lose their biochemical activity are administered to ruminants, they must thus be protected against this undesired microbial activity in the rumen. The literature describes various methods that can confer rumen protection to polyunsaturated fatty acids (PUFAs), such as CLAs. This is also evident from the findings of, for example, Elgersma et al. in Fresh Herbage for Dairy Cattle, 175 - 194, 2006, which describes the biochemical reduction rate of unsaturated fatty acids in the range of 82 - 98%. After passing through the rumen, only about 2 - 22% of the polyunsaturated fatty acids (PUFAs) that have been ingested can still be detected.

[0008] Therefore, the literature (see Kirchgeβner, see above) describes various forms of rumen-protected fats that resist microbial degradation and microbial modification:

[0009] Particularly mentioned are:

[0010] a) Naturally protected cell-bound oilseed fats, which are used in the form of oilseed cakes or ground whole seeds. The principle of protection here is to enable the slow release of oil from plant cells; however, the storage stability of such fats is poor.

[0011] b) Heat-treated oilseeds, such as by extrusion; here, the denatured proteins surround the fat droplets, thereby preventing their degradation by microorganisms.

[0012] c) Chemically modified fats, such as fat droplets encapsulated by saponifying fatty acids with calcium or proteins treated with formaldehyde.

[0013] d) Fats that prevent microbial enzymes by process methods (such as encapsulation with hardened vegetable fats).

[0014] EP-A-1 100 489 discloses a method for reducing the milk fat content of cows, in which an effective amount of CLA is administered to cows. As a way to prevent modification by rumen bacteria, it is proposed to administer the active substance by injection or provide it in coated form. Description of the Drawings

[0016] Figure 1 Shows the retention characteristics over time of various in vitro incubated CLA formulations of particles and extrudates (T1 and T3 of the present invention). Summary of the Invention

[0018] The object of the present invention is to provide a simplified route for converting rumen-unstable feed components into a dosage form with improved rumen stability.

[0019] Surprisingly, this object is achieved by providing a feed of uncoated particles according to the appended claims.

[0020] Specific Embodiments of the Invention

[0021] a) General Definitions

[0022] "Particles" or "particle form" includes solid feed products that can be obtained by known methods per se, with the aid of conventional granulation devices, and also conventional extrusion or expansion devices. Generally, such particles have a length of about 0.5 - 2 or 0.8 - 1.5 cm, for example about 1 cm, and a diameter in the range of 0.1 - 0.8 or 0.4 - 0.5 cm. These terms also include so-called "ground" particles, i.e., particles that are converted into smaller particles by the action of mechanical forces. The purpose of grinding is usually to make it easier for animals to ingest the particles, or otherwise to better distribute them in the final feed. Grinding allows for targeted reduction of the particle size and increase in the number of particles, for example, obtaining a particle mixture content > 50%, for example 55 - 95 or 60 - 90 or 70 - 80% of particles having a diameter in the range of about 3 - 6 or 4 - 5 mm.

[0023] "Rumen labile components" (pure substances and also mixtures of natural or synthetic substances) comprise chemical compounds which may undergo chemical modification (modification of the chemical empirical formula, e.g. by biochemical reduction, and / or configurational and / or stereochemical changes) when passing through the rumen of ruminants. Usually, these are organic chemical substances, in particular those which are of importance as food / feed components or food / feed additives. Those which must be mentioned in particular are saturated or mono- or polyunsaturated carboxylic acids, e.g. those having at least 6 carbon atoms, such as the CS, PUFA, MUFA or CLA described below. Further feed additives or feed components or single feeds (Einzelfuttermittel) which must / can be administered in a rumen-protected form are amino acids (in particular methionine, lysine etc.), enzymes, choline and other active substances from the vitamin group.

[0024] A "rumen labile mixture" or "rumen labile feed mixture" comprises at least one "rumen labile" component and is unprotected in the rumen or insufficiently rumen-protected, i.e. the component may be exposed to the chemical changes described above if fed and passed through the rumen of ruminants.

[0025] In the context of the present invention, "rumen stability" or "rumen protection" means that a "rumen labile" component is converted in the form of the above-described "granules" into a state of reduced "rumen lability" up to a state of substantially complete rumen protection. This improved rumen stability or reduced rumen lability can be determined in a simple manner by comparing the stability of the components in granulated or non-granulated mixtures using the test methods described in the test section (in vitro or in vivo).

[0026] "Carboxylic acids" (CS) are in particular straight-chain or branched-chain, in particular straight-chain saturated or mono- or polyunsaturated, optionally substituted C6-C 30 -monocarboxylic acids. Examples of saturated unbranched fatty acids are caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecanoic acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, palmitic acid, margaric acid, stearic acid, nonadecanoic acid, arachidic acid, behenic acid, lignoceric acid, cerotic acid and melissic acid. Examples of monounsaturated fatty acids are palmitoleic acid, oleic acid and erucic acid. Examples of diunsaturated fatty acids are sorbic acid and linoleic acid. Examples of triunsaturated fatty acids are linolenic acid and eleostearic acid. Examples of tetra- and polyunsaturated fatty acids are arachidonic acid, clupanodonic acid, eicosapentaenoic acid and docosahexaenoic acid. Examples of substituted fatty acids are ricinoleic acid ((R)-12-hydroxy-(Z)-9-octadecenoic acid). Further suitable fatty acids are naturally occurring fatty acids, such as gondoic acid and nervonic acid. If double bonds are present in the fatty acid, they can exist in cis and trans forms. The substituents are preferably selected from hydroxyl groups and lower alkyl groups such as methyl and ethyl. A keto group or an epoxy group can also be present in the hydrocarbon group, for example, in vernolic acid. Additional functional groups are cyclopropane, cyclopropene, and cyclopentene rings, which can be formed by bridging two adjacent carbon atoms in the hydrocarbon group of a fatty acid (e.g., and chaulmoogric acid).

[0027] "PUFAs" are polyunsaturated fatty acids having at least two conjugated or non-conjugated C═C double bonds in the fatty acid molecule. Examples that can be mentioned are: linolenic acid, eicosapentaenoic acid (EPA) ((5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenoic acid; or C20:5(ω-3)) and docosahexaenoic acid (DHA) ((4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid) or C22:6(ω-3)).

[0028] "MUFAs" are monounsaturated fatty acids, which can exist in cis or trans configurations, such as oleic acid or elaidic acid.

[0029] Conjugated linoleic acid (CLA) is a group of isomers of the C 18 -monocarboxylic acid "linoleic acid", which is diunsaturated and has its two double bonds in the 9 and 12 positions and is thus non-conjugated. Linoleic acid is also named by the abbreviation "C18:2 cis-9, cis-12".

[0030] "CLA" mainly includes all conjugated diunsaturated isomers of linoleic acid (C18:2 cis-9, cis-12), where the positions of the two double bonds on the carbon chain can be shifted towards the chain end or towards the carboxyl group, and where the stereochemistry of the conjugated double bonds can also include any alternative embodiments (cis / cis, trans / trans, cis / trans), where "cis / trans" includes two sequences "trans-cis" or "cis-trans", where in each case, the first-mentioned configuration of the two sequences refers to the double bond closest to the carboxyl group:

[0031] As an example, conjugated linoleic acid "C18:2 cis-9, trans-11" can be involved, as shown below:

[0032]

[0033] Therefore, the names cis / trans-9,11-linoleic acid, cis / trans-8,10-linoleic acid, cis / trans-11,13-linoleic acid, and cis / trans-10,12-linoleic acid used herein include cis-trans and trans-cis isomers, in other words:

[0034] Cis / trans-9,11-linoleic acid includes: C18:2 cis-9, trans-11 and C18:2 trans-9, cis 11

[0035] Cis / trans-8,10-linoleic acid includes: 18:2 cis-8, trans-10 and 18:2 trans-8, cis 10

[0036] Cis / trans-11,13-linoleic acid includes: 18:2 cis-11, trans-13 and 18:2 trans-11, cis 13

[0037] Cis / trans-10,12-linoleic acid includes: 18:2 cis-10, trans-12 and 18:2 trans-10, cis 12

[0038] Similarly, it is applied to linolenic acid, which is tri-unsaturated (C18:3, cis-9, cis-12, cis-15) and its cis / trans isomers.

[0039] The information on the above substances and the uses of these substances described herein mainly relate to the corresponding pure substances, and also to natural or synthetic mixtures of substances that contain at least one of these substances, such as at least one PUFA or at least one CLA.

[0040] Natural mixtures of substances are, for example: fish oil or microbial oil, which can be rich in PUFAs; and linseed oil, soybean oil, sunflower oil, castor oil, etc.

[0041] Synthetic mixtures of substances are, for example, commercially available products rich in CLA, such as those of BASF SE

[0042] Other substances that must be mentioned are rumen-unstable derivatives of the above carboxylic acids (CS, PUFA, MUFA, CLA), for example, especially substituted derivatives. Examples that must be mentioned are compounds that are mono- or poly-substituted, for example by hydroxyl groups, on the hydrocarbon radical of the carboxylic acid. Examples of such compounds that must be mentioned are: 10-hydroxy-cis-12-octadecadienoic acid.

[0043] b) Specific embodiments

[0044] The present invention particularly relates to the following embodiments:

[0045] 1. A particulate ruminant feed, in particulate form (i.e., relatively small particles obtained by granulation, extrusion, expansion, and also in comminuted form, i.e., obtained by subsequently comminuting the granules, extrudates or expanded materials), the particulate ruminant feed comprising a mixture of at least one solid particulate feed component and at least one rumen labile component, said rumen labile component having been added to the mixture (in pure form or as a component of a natural or synthetic mixture of substances) (rumen labile), wherein, in particular, each, several or all of the mixture components used for granulation together with the rumen labile component do not themselves exhibit any or insufficient rumen stabilizing activity in the granulated mixture. In particular, for example, in a granulated mixture containing CLA, such mixture components are completely absent or present only in ineffective amounts, which bring about rumen stabilization of CLA in a non-particulate CLA mixture. Examples that must be mentioned in the context are additives having stabilizing activity, which are present in commercially available rumen-stabilized CLA products such as, for example, especially soybean oil, gypsum or silica, which are present in the feed mixtures of the present invention either not granulated at all or only in non-stabilizing amounts.

[0046] The mixture for preparing the feed particles can thus be called a "rumen labile" mixture, which is only in comparison with "rumen stability", i.e., its rumen lability is reduced by the granulation, extrusion or expansion step of the present invention.

[0047] As a result, in a preferred case of the present invention, no feed mixture is processed into particles having sufficient rumen stability before granulation. For example, the present invention thus does not include processing a mixture of rumen-protected components (such as a CLA mixture or a mixture of equivalent compositions) into particulate ruminant feed.

[0048] 2. The feed according to embodiment 1, wherein the component is a fat or an oil, especially a fatty acid (a carboxylic acid having at least 6 carbon atoms).

[0049] 3. The feed according to any one of the above embodiments, wherein the amount of the added rumen labile component ranges from 0.1 - 20, 1 - 15, 2 - 10 or 3 - 5% by weight of the total weight of the granulating mixture.

[0050] 4. The feed according to any one of the above embodiments, wherein the component is selected from saturated and mono- or poly-unsaturated (especially di-unsaturated) carboxylic acids having at least 6 carbon atoms, mixtures of at least two saturated and / or mono- or poly-unsaturated (especially di-unsaturated) carboxylic acids having at least 6 carbon atoms, and substance mixtures comprising at least one saturated or mono- or poly-unsaturated (especially di-unsaturated) carboxylic acid having at least 6 carbon atoms, such as C18:2 or C18:3 carboxylic acids; also included are substituted derivatives of such carboxylic acids, such as hydroxy-substituted derivatives, such as 10-hydroxy-cis-12-octadecadienoic acid.

[0051] 5. The feed according to any one of the above embodiments, wherein the component comprises at least one poly-unsaturated (especially poly-unsaturated) fatty acid (PUFA), especially having conjugated double bonds.

[0052] 6. The feed according to embodiment 5, wherein the poly-unsaturated (especially di-unsaturated) fatty acid comprises at least one isomer of linoleic acid.

[0053] 7. The feed according to embodiment 6, wherein the poly-unsaturated fatty acid is CLA.

[0054] 8. The feed according to embodiment 7, wherein CLA is selected from:

[0055] a) cis / trans-9,11-linoleic acid;

[0056] i.e., C18:2 cis-9, trans-11 and 18:2 trans-9, cis 11

[0057] b) cis / trans-8,10-linoleic acid;

[0058] i.e., C18:2 cis-8, trans-10 and 18:2 trans-8, cis 10

[0059] c) cis / trans-11,13-linoleic acid;

[0060] i.e., C18:2 cis-11, trans-13 and 18:2 trans-11, cis 13

[0061] d) cis / trans-10,12-linoleic acid;

[0062] i.e., C18:2 cis-10, trans-12 and 18:2 trans-10, cis 12

[0063] e) cis / cis-9,11-linoleic acid;

[0064] f) trans / trans-9,11-linoleic acid;

[0065] g) cis / cis-8,10-linoleic acid;

[0066] h) trans / trans-8,10-linoleic acid;

[0067] i) cis / cis-11,13-linoleic acid;

[0068] j) trans / trans-11,13-linoleic acid;

[0069] k) cis / cis-10,12-linoleic acid;

[0070] l) trans / trans-10,12-linoleic acid; and mixtures of at least two of the compounds mentioned above.

[0071] 9. The feed according to embodiment 8, wherein the CLA is selected from:

[0072] a) 9-cis,11-trans-linoleic acid (C18:2 cis-9, trans-11);

[0073] b) 10-trans,12-cis-linoleic acid (C18:2 trans-10, cis 12); and

[0074] c) mixtures thereof.

[0075] 10. A method for preparing a ruminant feed in the form of granules according to any one of the above embodiments, wherein

[0076] a. Mixing at least one solid particulate feed component with at least one rumen labile ingredient as defined above;

[0077] b. Optionally treating the resulting mixture, in particular by introducing steam, in particular treating with hot (e.g. up to 95 °C) steam, such as saturated steam, for a sufficient time period, such as 20 - 40 or 20 - 120 or 20 - 140 or 20 - 240 seconds;

[0078] c. Pressing the optionally treated mixture under pressure in an extrusion, expansion or granulation device, in particular a granulation or extrusion device, to obtain granules; and

[0079] d. Optionally cooling and / or subjecting the resulting granules to a final drying process.

[0080] 11. A method for altering the milk fat concentration in milk produced by a lactating ruminant, wherein the lactating ruminant is provided with an effective amount of the ruminant feed in the form of granules according to any one of embodiments 1 - 9 or prepared according to embodiment 10.

[0081] 12. The method according to embodiment 11, wherein the lactating ruminant is selected from cows, goats and sheep.

[0082] 13. Milk prepared by the method according to embodiment 11 or 12.

[0083] 14. A method for preparing a rumen-protected ruminant feed for rumen-labile components, wherein

[0084] a. at least one solid particulate feed component is mixed with at least one rumen-labile component to obtain a rumen-labile mixture of said component and at least one feed component (i.e., a mixture that does not show or shows only insufficient rumen protection of the labile component);

[0085] b. the resulting mixture is optionally treated;

[0086] c. in a granulation, extrusion or expansion device, the optionally treated mixture is pressed by the action of pressure and optionally steam to obtain pellets; wherein the mixture of pellets shows improved rumen stability of the components compared to the non-pelleted mixture (i.e., shows a reduction in the rumen lability of said components to preferably substantially complete rumen protection); and

[0087] d. optionally cooling and / or subjecting the pellets thus obtained to a final drying process and optionally subsequent grinding.

[0088] 15. A feed according to one of embodiments 1-9, prepared by the method according to embodiment 14.

[0089] d) Further embodiments

[0090] d1) Preparation of feed pellets

[0091] To prepare a pelleted feed composition, a rumen-labile component formulated according to the invention is mixed, optionally together with additional conventional animal feed components (such as a milk-producing feed) added. The amount of the component is selected such that it ranges, for example, from 0.1-20, 1-15, 2-10 or 3-5% by weight. Thereafter, the feed is pelletized by means of a suitable granulator. For this purpose, the feed mixture is usually treated with steam (hot steam with a temperature up to 95 °C, such as saturated steam) for, for example, 20-1240 seconds. The temperature increase during granulation can be controlled by the amount of saturated steam added. However, this steam treatment can also be omitted.

[0092] Pelletization is usually carried out in a commercially available granulator (e.g., from Bühler GmbH), which can be designed as a ring die press. Here, the roller presses the pre-treated material mixture through a ring die (the gap width between the roller and the die is, for example, 1 mm). Depending on the die, pellets with a diameter of approximately 2 - 12 mm can be prepared. For dairy cattle feed, a die with an inner diameter of, for example, 4 - 5 mm and a slot length of 40 - 50 mm is applied. When the mixture is pressed through the die, the maximum processing temperature prevails. In this stage, a temperature range of approximately 60 - 100 °C can be reached.

[0093] The hot material remaining at the die outlet is continuously cut with a blade, and the pellets thus formed are immediately cooled with a cooler, and an optional final drying process is carried out. After cooling and optional drying, the residual water content range can be approximately 1 - 15% by weight of the total pellet weight.

[0094] Alternatively, granulation can be replaced by extrusion or expansion (e.g., using a feed expander from Kahl, Germany) (see, for example, “Feed Manufacturing Technology IV” edited by McEllhiney, Kansas State Univ., Pub., American Feed Industry Assoc, 1994; Arlington, VA., where it can be found: Extrusion Cooking Systems, Bob Hauck et al., pages 131 - 139; Wilson et al., 1998, Journal of Poultry Science, 77 (Suppl 1): 41). The products in these processes can then be crushed.

[0095] d2) Application

[0096] Typical feed components that can be used to produce the granules of the present invention include single feeds of plant or animal origin according to the Futtermittelverordnung (FMV; German Feed Regulation), such as cereal by-products, fine wheat bran, wheat bran; extracted flours, apple pomace, dried molassed beet pulp, fish meal, meat and bone meal; and / or single mineral feeds according to the FMV, such as carbonates, phosphates, sulfates, propionates. Other suitable feeds are cereals, such as wheat, rye, barley, oats, maize, millet or triticale; cereal by-products (ground by-products), such as bran, feed, wheat feed, fine bran or wheat semolina; by-products in oil production (extracted flours, expellers, cakes); by-products in sugar production (molasses, dried beet pieces, sugar for feed, pulp, potato starch, corn gluten, wheat gluten); by-products of the fermentation industry, brewery grains, yeast, malt, stillage; and foods of animal and other origins, such as blood meal, fish meal, pressed grape juice, potato protein.

[0097] Those that must be mentioned in particular are wheat, grain maize, barley, oats; soybeans; cereal flours, such as wheat or maize flour, soybean flour, molassed beet pieces, wheat feed, wheat semolina, corn gluten feed, extracted soybean flour, extracted rapeseed flour, brewery grains, cereal stillage, beet molasses, oat bran; sugars, such as glucose and sugar alcohols, as well as protein components, such as soybean concentrate, fish meal, gluten such as corn or wheat gluten, oils and fats, and nutrient products, such as free amino acids, their salts, vitamins (such as A, D, E) and trace elements (such as Cu, as CuSO4), mineral components, such as calcium carbonate, sodium chloride; phosphates and optionally processing aids, such as flow aids, inert fillers, etc.; and optionally preservatives.

[0098] Typical milk-producing feed compositions include, for example, maize, wheat, barley, oats, rye, citrus apple pomace, extracted soybean flour, extracted rapeseed food, soybean hulls, palm kernel expeller, DDGS, corn gluten feed, beet pieces, wheat feed, wheat bran, linseed, molasses, lime, salt, vitamin and trace element premixes.

[0099] The present invention is now illustrated more specifically with reference to the following application examples.

[0100] Experimental section

[0101] Example 1: Study on the reduction of milk fat when dairy cows are fed with CLA feed formulations of multiple particles

[0102] a) Granulation

[0103] Granulator: Manufacturer Simon Heessen, Type V3-30C.

[0104] Capacity: 600 - 650 kg / hour

[0105] Mold:

[0106] Inner diameter: 5 mm

[0107] Internal length: 45 mm

[0108] Clearance width: 1 mm

[0109] Pelletizing temperature: 80 °C

[0110] Treatment: Steam T = 135 and 145 °C (depending on pressure)

[0111] b) The CLA formulation used

[0112] (1) (Commercially available product containing CLA)

[0113] Table: Compositions of 5 different batches

[0114]

[0115]

[0116] 1 GC area %

[0117] (2)

[0118] Lutrell consists of 34% Lutalin and additives 15% silica, 5% gypsum and 46% hydrogenated soybean oil, where the rumen stabilization function is mainly affected by the latter.

[0119] The composition of Lutrell (fatty acid distribution) is shown in the following table.

[0120]

[0121] (3) Lutalin + Silafett

[0122] The fatty acid composition of Silafett = hydrogenated soybean oil

[0123]

[0124] b) Experimental design

[0125] Test 4 experimental schemes (see the table below).

[0126] Scheme 1 (T1) is the control (C) and does not contain any CLA;

[0127] Protocol 2 (T2) contains the CLA formulation "Lutrell";

[0128] Protocol 3 (T3) contains the CLA formulation "Lutalin" and

[0129] Protocol 4 (T4) contains the CLA formulation "Lutalin + Silafett".

[0130] T1 T2 T3 T4 Soybean / corn mixture <![CDATA[1000 1) > 950 983.3 950.0 Lutrell 0 50 0.0 0.0 Lutalin 0 16.7 16.7 Lutalin + Silafett 0 0.0 33.3 1000 1000 1000 1000

[0131] 1) Data in grams

[0132] Feed: 1 kg of supplement consists of 25% pre - granulated extracted soybean meal and 75% corn meal.

[0133] The CLA product is incorporated into the lactating feed (25% by weight of extracted soybean meal; 75% by weight of corn meal) and the mixture is granulated.

[0134] The experimental design is equivalent to a 4×4 Latin square (see the table below). In the experimental groups, in each case, pure CLA is dosed at the same amount of 16.7 g per cow per day.

[0135]

[0136]

[0137] Designate the results in the table below (part f): C = control = T1, Lutrell = T2, Lutalin = T3, and Lutalin + Silafett = T4.

[0138] Each period is 21 days; i.e., the adaptation period is 14 days, followed by a 7 - day measurement period. All 4 experimental feeds are based on the same basic ingredients.

[0139] c) Animals

[0140] Test each treatment according to the experimental design for 20 cows described above. Designate another 4 cows into the experimental group as reserve animals. At the start of this experiment, the experimental cows were in the mid - lactation segment. The average milk production at the start of this experiment was approximately 32 kg FCM / day. The experimental cows (German Holstein) were selected from the herd at the experimental station (herd performance about 11000 kg milk, 3.9% fat, 3.4% protein) and designated into 4 groups according to the following criteria,

[0141] · Number of lactations (1 and above)

[0142] · Milk fat content (based on two prior milk production records)

[0143] · Days in lactation and milk production

[0144] The aim is to adjust comparable group means.

[0145] d) Feeding

[0146] All cows were provided with the same total mixed ration (TMR), which was fed once daily at random. The TMR consisted of cereal silage, grass silage, hay and approximately 45% concentrate. The TMR target values are shown in the table below. This TMR ensures that if a cow consumes approximately 21 kg DM of this feed, it is provided with sufficient nutrients for 33 kg of milk production.

[0147]

[0148] Crude nutrient content of the TMR according to Weender analysis (uXP = available crude protein, XP = crude protein, XL = crude fat, XF = crude fiber, and NEL = net energy for lactation).

[0149] First, three CLA formulations were used to prepare a premix with the pellets of the milk-producing feed. Then, in the feed trough, an appropriate amount of these premixes (equivalent to 1 kg / animal / day) was manually mixed with the TMR, aiming to obtain 16.7 g of the pure substance in 20 kg DM of TMR in each case. The control group received the same amount of the same concentrate but without CLA.

[0150] e) Sampling and measured parameters

[0151] Feed

[0152] To determine the dry matter, TMR samples were taken once daily and combined to obtain a cumulative sample for each measurement period (7 days). This cumulative sample was used to determine the crude nutrient content according to Weender analysis and the energy content according to the Hohenheimer feed value test.

[0153] Cows

[0154] For each cow, the (respective) TMR intake, milk yield and live weight were recorded daily. During the 7-day measurement period, three equal aliquots of milk samples were taken from the evening and morning milk, and the fat, total protein (Nx6.38), lactose and urea contents were tested, and the somatic cell count was tested by the Baden-Württemberg Milk Testing Institute. Additional milk tests (preparation period) were carried out in the middle of the first and second weeks of each test period. Cumulative milk samples containing the 7-day measurement period were reconstituted for each cow and each treatment and then frozen to enable optional fatty acid analysis of the milk fat therein.

[0155] f) Test results

[0156] Table: Effects of Lutalin and Lutrell in the form of feed for granules on the reduction of milk fat in dairy cows fed 5 g of trans-10 / cis-12 CLA

[0157]

[0158]

[0159] SD: Standard deviation

[0160] Surprisingly, treatments T3 and T4 resulted in the same reduction of milk fat as treatment T2, although the active substance CLA in T3 and T4 had been incorporated into the lactating feed in a non- rumen-protected form. Thus, granulation protected the active substance CLA in T3 and T4 in the rumen to the same extent as known from Lutrell and was also confirmed by T2.

[0161] Example 2: Study on the rumen stability of CLA feed formulations of different particles

[0162] Stability in the rumen is essential for the reduction of milk fat of feeds containing CLA. For this purpose, the feeds tested in Example 1 (75% corn + 25% extracted soybean meal) were mixed with 1.66% Lutalin or 5% Lutrell, and the mixtures were granulated or extruded. Then the resulting granules and extrudates were incubated in vitro for 4, 12, and 24 hours. After the predetermined time, the incubation was stopped, and the entire content was transferred to a container and freeze-dried. Subsequently, the CLA content was determined for this freeze-dried sample material.

[0163] a) Granulation and extrusion

[0164] Preparation of powder: Corn and extracted soybean meal were weighed together, ground, and mixed in a horizontal mixer. Lutalin or Lutrell was added and mixed. The total mixing time was 15 minutes. In each case, the two mixtures were separated. 80 kg was granulated and 30 kg was extruded.

[0165] Granulator: Manufacturer Simon Heesen, horizontal die 3.3×35 mm, rated throughput 300 kg / hour.

[0166] Extruder: Werner & Pfleiderer 37, die Rated throughput up to 50 kg / hour.

[0167] Granulation: Granulation temperature reached 67 - 68.5 °C.

[0168] Extrusion: The extrusion temperature reached 85 °C and about 14% water had been added. A residual water content of 10 - 12% was found in the final product.

[0169] b) CLA formulation used

[0170] (1) (Commercially available product containing CLA. Composition see Example 1.

[0171] (2) Pure: Commercially available product containing CLA. Composition see Example 1.

[0172] c) Test setup

[0173] Four test scenarios were tested (see table below).

[0174] Scenario 1 (T1): 1 type of granule and containing the CLA formulation "Lutalin"

[0175] Scenario 2 (T2): 1 type of granule and containing the CLA formulation "Lutrell"

[0176] Scenario 3 (T3): 1 type of extrudate and containing the CLA formulation "Lutalin"

[0177] Scenario 4 (T4): 1 type of extrudate and containing the CLA formulation "Lutrell"

[0178] T1 T2 T3 T4 Soybean / corn mixture 983.3* 950 983.3 950.0 Lutrell 50 0,0 50 Lutalin 16.7 16.7 0 Total 1000 1000 1000 1000

[0179] * Data in grams

[0180] Feed: 1 kg of supplement consists of 25% extracted soybean meal and 75% corn meal.

[0181] Analysis carried out immediately after the processing step showed that the active substance CLA kept the granulation process extremely good, while extrusion led to an activity loss of about 40%.

[0182] d) Incubation: Determination of in vitro degradation products in the Hohenheimer feed value test (in vitro test)

[0183] The Hohenheimer feed value test (HFT) is an in vitro method for evaluating the energetic feed value of ruminant feeds. When incubation is stopped after a defined time period and the residues of the active substances in the incubation solution are analyzed, suitable scenarios can also test the rumen stability of the active substances in the HFT.

[0184] The Hohenheimer feed value test (HFT) is carried out according to the provisions in the VDLUFA method (Methodenbuch [Method Book], Volume III, Chapter 25.1).

[0185] The incubation period depends on the purpose of the test; conveniently, it is 4 - 24 hours due to the degradation of the CLA formulation. The in vitro system can be stable for up to 48 hours; however, beyond this time point, deviations in the physiological process of fermentation must be anticipated. In each case, weigh 500 mg of test protocols T1 - T4 (equivalent to approximately 5 mg of CLA or approximately 3 mg for the extrusion protocol). The incubation periods total 4, 12, and 24 hours. For each incubation period, three determinations are made (3 incubation vessels for each test protocol T1 - T4).

[0186] After the relevant period, terminate the incubation by cooling with ice - water. Transfer the rumen fluid / buffer mixture quantitatively into glass containers, which are suitable for the subsequent freeze - drying step. Weigh the empty drying containers so that after drying, the residual weight can be determined and the data used for subsequent quantitative analysis. The freeze - drying system used is the Gamma 1 - 20 from Christ, 37507 Osterode.

[0187] e) Analytical method: Improvement of AM / 00887 / 01

[0188] Due to the specific matrix and low CLA content, the processing as described in the analytical method (see Chapter 7.4.2) must be improved as follows. The improvements made are shown in italics.

[0189] Weigh the sample as described in 7.4.2 or introduce it into a 250 - mL Erlenmeyer flask; thereafter, add the stated amounts of BHT, sodium ascorbate, and water.

[0190] Then, add a heaping spatula tip of the enzyme Pronase, and place the Erlenmeyer flask in an ultrasonic bath at 60 °C for 15 minutes.

[0191] After adding the stated amounts of ethanol and acetic acid as described in 7.4.2, allow the sample to return to the ultrasonic bath at 60 °C for 15 minutes.

[0192] After cooling to room temperature, add the extraction solution cyclohexane / ethyl acetate 80:20 (v / v). However, only use 50 mL of the extraction solution instead of the usual 100 mL.

[0193] Thereafter, stir the sample with a magnetic stirrer for 30 minutes as described in 7.4.2, add 70 mL of saturated sodium chloride solution, and continue stirring for 10 minutes. Thereafter, turn off the stirrer and keep the mixture static until phase separation occurs.

[0194] 3 mL of the organic phase was taken, and the solvents cyclohexane and ethyl acetate were removed in a nitrogen stream, and the residue was dissolved in 2 mL of the extraction solution cyclohexane / ethyl acetate 80:20 (v / v) (concentrated).

[0195] The resulting sample solution aliquots were directly filtered through a disposable 0.45-μm filter into HPLC vials.

[0196] The injection volume was 20 μL instead of the usual 10 μL.

[0197] f) Test results

[0198] Total CLA retention (%) (as methyl esters and free fatty acids)

[0199]

[0200] In this in vitro test, the same milk fat reduction of Lutalin particles compared to Lutrell particles, which was surprisingly found in Example 1, could be reproduced, since the CLA retention measured in both particle formulations T1 and T2 was the same. Thus, using this test, it was possible to at least definitely evaluate those formulations that were based on the same feed but differed only in the physical treatment. Surprisingly, the extrusion formulations T3 and T4 showed in each case a retention percentage that was substantially higher than that of the particle formulations - after 4 and 12 hours and after 12 and 24 hours, respectively (see Figure 1 ).

[0201] Therefore, it can be concluded that the extruded form of Lutalin (non-rumen-stable CLA) also has at least comparable rumen stability to a comparable Lutrell extrudate (CLA that has been rumen-stabilized with a stabilizing additive).

[0202] Reference is made in particular to the disclosures of the publications mentioned herein.

Claims

1. A particulate ruminant feed, which is in particulate form, the particulate ruminant feed comprising a mixture of at least one solid particulate feed component and at least one rumen labile component, said rumen labile component having been added to the mixture.

2. The feed of claim 1, wherein said component is a fat or an oil.

3. The feed of any of the preceding claims, wherein the amount of rumen labile component added ranges from 0.1 - 20% by weight of the total weight of the granulating mixture.

4. The feed of any of the preceding claims, wherein said component is selected from saturated and mono- or poly-unsaturated carboxylic acids having at least 6 carbon atoms, mixtures of at least two saturated and / or mono- or poly-unsaturated carboxylic acids having at least 6 carbon atoms, and mixtures of substances comprising at least one saturated or mono- or poly-unsaturated carboxylic acid having at least 6 carbon atoms.

5. The feed of any of the preceding claims, wherein said component comprises at least one polyunsaturated fatty acid, especially having conjugated double bonds.

6. The feed of claim 5, wherein said polyunsaturated fatty acid comprises at least one isomer of linoleic acid.

7. The feed of claim 6, wherein said polyunsaturated fatty acid is CLA.

8. The feed of claim 7, wherein CLA is selected from: a) cis / trans-9,11-linoleic acid; b) cis / trans-8,10-linoleic acid; c) cis / trans-11,13-linoleic acid; d) cis / trans-10,12-linoleic acid; and e) mixtures of at least two of the compounds mentioned above.

9. The feed of claim 8, wherein CLA is selected from: a) 9-cis,11-trans-linoleic acid; b) 10-trans,12-cis-linoleic acid; and c) mixtures thereof.

10. A method for preparing a particulate ruminant feed of any of the preceding claims, wherein a. mixing at least one solid particulate feed component with at least one rumen labile component; b. optionally treating the resulting mixture; c. pressing the optionally treated mixture in a granulating, extruding or expanding device by means of pressure and optionally steam action to obtain particles; and d. optionally cooling and / or subjecting the resulting particles to a final drying process and optionally subsequently comminuting.

11. A method for preparing a rumen-protected ruminant feed, said rumen-protected ruminant feed comprising at least one rumen labile component, wherein a. mixing at least one solid particulate feed component with at least one rumen labile component to obtain a rumen labile mixture of said component and at least one feed component; b. optionally treating the resulting mixture; c. pressing the optionally treated mixture in a granulating, extruding or expanding device by means of pressure and optionally steam action to obtain particles; wherein the mixture of particles shows improved rumen stability of said component (reduced rumen lability of said component) compared to the non-particulate mixture; and d. optionally cooling and / or subjecting the particles thus obtained to a final drying process and optionally subsequently comminuting.

12. A method for altering the milk fat concentration in milk produced by a lactating ruminant, wherein the lactating ruminant is provided with an effective amount of a ruminant feed comprising the particles of any one of claims 1-9 or prepared according to claim 10.

13. The method of claim 12, wherein the lactating ruminant is selected from cows, goats, and sheep.

14. Milk produced by the method according to claim 12 or 13.

15. A feed according to any one of claims 1-9 prepared by the method of claim 11.

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