PELLETED RUMINANT FEED ENRICHED WITH RUMEN-LABILITY INGREDIENTS

Description

[0001] The invention relates to pelleted ruminant feed containing, in pelleted form, a mixture of at least one solid particulate feed ingredient with at least one rumen-unstable ingredient added to the mixture; to processes for their production and to their use in methods for altering the milk fat concentration. Background of the invention

[0002] Additives to ruminant feed can be subject to undesirable chemical changes by rumen organisms. For example, during the modification of polyunsaturated fatty acids by rumen microbes, both the content and the position of double bonds in the carbon chain of the fatty acid can change. Typically, the double bonds of unsaturated fatty acids in plants are separated by at least two single bonds. Microbial enzymes can generate a conjugated double bond system in which the double bonds are separated by only one single bond. These include, for example, conjugated linoleic acid isomers (CLA). They are primarily produced by the linoleic acid isomerase of the rumen bacteria. Butyrivibrio fibrisolvensfrom linoleic acid (C18:2 cis-9, trans-12). The main CLA isomer is cis-9, trans-11 CLA (C18:2 cis-9, trans-11), accounting for approximately 75–92% of the total CLA content. In contrast, the milk fat-depressing isomer C18:2 trans-10, cis-12 accounts for only a small proportion of 0.03–1.5%. A portion of these isomers enters the ruminant body via absorption. In dairy cows, the rumen-modified fatty acids are also transported into the milk. The CLA content of milk fat is highly dependent on feeding and ranges from approximately 0.3–1.1%. (cf. Kirchgeßner, Tierernährung, 13th edition, DLG Verlag GmbH), and in pasture-fed animals, up to over 4% (Tröscher et al., 2014; in press). CLAs are being intensively researched due to their potent physiological and pharmacological effects. In cell culture studies and animal experiments, CLAs demonstrate cancer-protective and anti-inflammatory effects.It is also known that conjugated linoleic acid isomers (CLA isomers) in ruminants inhibit the expression of numerous genes responsible for the uptake of circulating fatty acids into the mammary gland, for fatty acid synthesis, and for triglyceride formation. The C18:2 trans-10, cis-12 CLA isomer, in particular, has proven to be particularly effective in reducing fat. In connection with the reduction in the fat content in milk, benefits discussed include relief of metabolic stress and an increase in milk yield. It has been reported that reductions in milk fat content of 0.4–0.5% can lead to increases in milk yield of up to 3–10% (cf. Kirchgeßner, op. cit.).

[0003] For this reason, CLAs are often added to dairy feed today. These preparations are permitted for use in feed, but must be used in a rumen-protected form, as unprotected CLAs are further broken down in the rumen into ineffective C18:1 and C18:0 fatty acids.

[0004] If these or other molecules that are degraded by rumen microorganisms or chemically altered to the point of losing their biochemical activity are to be administered to ruminants, they must be protected against such undesirable microbial effects in the rumen. Various methods have been described in the literature that can confer rumen protection on polyunsaturated fatty acids (PUFAs), such as CLAs. This is also illustrated by findings, for example, by Elgersma et al., in Fresh Herbage for Dairy Cattle, 175-194, 2006, which describe biohydrogenation rates of unsaturated fatty acids in the range of 82-98%. Only about 2 to 22% of the ingested polyunsaturated fatty acids (PUFAs) are still detectable after passage through the rumen.

[0005] Thus, various forms of rumen-protected fats that are protected against microbial degradation and microbial modification are described in the literature (cf. Kirchgeßner, ibid.): In particular, the following are mentioned: a) Natively protected cell-bound oilseed fats, which are used in the form of oilseed cakes or crushed whole seeds. The principle of protection lies in the slow release of oils from the plant cells; however, such fats have a short storage stability. b) Heat-treated oilseeds, e.g., by extrusion; denatured proteins surround the fat droplets and thus protect them from microbial degradation. c) Chemically modified fats, e.g., by saponification of fatty acids with calcium or coating of the fat droplets with formaldehyde-treated proteins. d) Fats that are protected from microbial enzymes by technical processes (e.g., coating with hydrogenated vegetable fats).

[0006] EP-A-1 100 489 discloses a method for reducing milk fat content in dairy cattle, whereby an effective amount of CLA is administered to the cattle. To protect against rumen bacterial changes, it is proposed to administer the active ingredient by injection or to provide it in coated form. Character description

[0007] Figure 1 shows the temporal retention of various in vitro incubated CLA formulations in pellets and extrudates (T1 and T3 according to the invention) Summary of the invention

[0008] The object of the invention is to provide a simplified way of converting rumen-unstable feed ingredients into a dosage form with improved rumen stability.

[0009] Surprisingly, the problem could be solved by providing uncoated, pelleted feeds according to the appended claims. Special embodiments of the invention a) General definitions

[0010] A "pellet" or a "pelleted form" encompasses solid feed formulations, such as those obtainable using conventional pelleting devices, as well as conventional extrusion or expansion devices in a manner known per se. Such pellets typically have a length of approximately 0.5 to 2 or 0.8 to 1.5 cm, such as approximately 1 cm, and a diameter in the range of 0.1 to 0.8 or 0.4 to 0.5 cm. These terms also include so-called "crumbled" pellets, i.e., pellets that are converted into smaller particles by mechanical force. The purpose of crumbling is often to facilitate the ingestion of the pellets by the animal or to improve their distribution in the final feed. By crumbling, the particle size can be specifically reduced and the particle number increased. This results, for example, in a particle mixture with a proportion of >50%, e.g.55 to 95 or 60 to 90 or 70 to 80% of particles with a diameter in the range of about 3 to 6 or 4 to 5 mm.

[0011] A "rumen-unstable ingredient" (pure substances but also natural or synthetic mixtures) comprises a chemical compound that, without adequate protection, can undergo chemical changes (changes in the chemical formula, such as through biohydrogenation, and / or in the configuration and / or stereochemistry) upon passage through the ruminant rumen. This is usually an organic chemical substance, particularly one that is important as a food / feed component or food / feed additive. Particular mention should be made of mono- or polyunsaturated carboxylic acids, such as those with at least 6 carbon atoms, such as the CS, PUFA, MUFA, or CLA listed below. Other feed additives, feed components, or feed materials that must / can be administered in a rumen-protected manner include amino acids (especially methionine, lysine, etc.), enzymes, choline, and other active ingredients from the vitamin class.

[0012] A "rumen-unstable mixture" or "rumen-unstable feed mixture" comprises at least one "rumen-unstable" ingredient and is not or insufficiently rumen-protected, i.e. the ingredient would be exposed to chemical changes as described above when fed to the animal and passing through the rumen.

[0013] "Rumen stability" or "rumen protection" in the context of the invention means that a "rumen-unstable" ingredient is converted into a state of reduced "rumen instability" through the above-described "pellet" form, up to and including essentially complete rumen protection. This improved rumen stability or reduced rumen instability can be easily determined by comparing the stability of the ingredient in a pelleted or non-pelletized mixture using experimental approaches (in vitro or in vivo) described in the Experimental Section.

[0014] "Carboxylic acids" (CS) are, in particular, straight-chain or branched, especially straight-chain, saturated or mono- or polyunsaturated, optionally substituted C 6 -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 four- 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). Other suitable fatty acids are naturally occurring fatty acids such as gondoic acid and neronic acid. If the fatty acids contain double bonds, these can be in either cis or trans form. The substituents are preferably selected from hydroxyl and lower alkyl groups, such as methyl and ethyl groups. Furthermore, keto groups or epoxy groups, such as in vernolic acid, can be present in the hydrocarbon radical. Other functional groups are cyclopropane, cyclopropene, and cyclopentene rings, which can be formed by bridging two adjacent carbon atoms in the hydrocarbon radical of the fatty acid (cf. malvalic acid and chaulmoogra acid).

[0015] "PUFA" are polyunsaturated fatty acids with at least two conjugated or non-conjugated C=C double bonds in the fatty acid molecule. Examples include linolenic acid, eicosapentaenoic acid (EPA) ((5 Z ,8 Z ,11 Z ,14 Z ,17 Z )-eicosa-5,8,11,14,17-pentaenoic acid; or C20:5 (ω-3) ) and docosahexaenoic acid (DHA) ((4 Z ,7 Z ,10 Z ,13 Z ,16 Z ,19 Z )-docosa-4,7,10,13,16,19-hexaenoic acid) or C22:6 (ω-3)).

[0016] "MUFA" are monounsaturated fatty acids that occur in cis or trans configuration, such as oleic acid or vaccenic acid.

[0017] Conjugated linoleic acids (CLA) are a group of isomers of the diunsaturated C18 monocarboxylic acid "linoleic acid," whose two double bonds are located at positions 9 and 12 and are therefore non-conjugated. This is also referred to by the abbreviation "C18:2 cis-9, cis-12."

[0018] "CLA" basically includes all conjugated, diunsaturated isomers of linoleic acid (C18:2 cis-9, cis-12), where the position of the two double bonds in the carbon chain can be shifted towards the chain end or towards the carboxyl group, and the stereochemistry of the conjugated double bonds can include any variation (cis / cis, trans / trans, cis / trans), where "cis / trans" includes both sequences "trans-cis" or "cis-trans", with the first configuration of the two sequences relating to the double bond closest to the carboxyl group:

[0019] As an example, reference is made to the conjugated linoleic acid "C18:2 cis-9, trans-11" shown below:

[0020] The terms used herein: 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; thus include both the cis-trans and the trans-cis isomers, i.e.: cis / trans-9,11-linoleic acid includes: C18:2 cis-9, trans-11 and C18:2 trans-9,cis 11 cis / trans-8,10-linoleic acid includes: C18:2 cis-8, trans-10 and C18:2 trans-8,cis 10 cis / trans-11,13-linoleic acid includes: C18:2 cis-11, trans-13 and C18:2 trans-11,cis 13 cis / trans-10,12-linoleic acid includes: C18:2 cis-10, trans-12 and C18:2 trans-10,cis 12

[0021] The same applies to triunsaturated linolenic acid (C18:3, cis-9, cis-12, cis-15) and the cis / trans isomers thereof.

[0022] The above substance information and the uses of these substances described herein primarily relate to the respective pure substance but also to natural or synthetic mixtures of substances which contain at least one of these substances, e.g. at least one PUFA or at least one CLA.

[0023] Natural mixtures of substances include fish oils or microbial oils, which can be rich in PUFAs, as well as linseed oil, soybean oil, sunflower oil, castor oil, etc.

[0024] Synthetic mixtures of substances include, for example, CLA-rich commercial products such as Lutalin ®< from BASF SE.

[0025] Furthermore, rumen-unstable derivatives of the above-mentioned carboxylic acids (PUFA, MUFA, CLA), especially substituted derivatives, should be mentioned. For example,

[0026] Compounds that are mono- or polysubstituted on the hydrocarbon residue of the carboxylic acid, such as by hydroxyl groups. An example of such a compound is 10-hydroxy-cis-12-octadecadienoic acid. b) Special embodiments

[0027] The present invention is described in the appended set of claims.

[0028] The invention therefore relates to a pelleted ruminant feed for dairy cattle, containing in uncoated, pelleted form (i.e. by pelleting, extrusion, expansion but also in crumbled form, i.e. by subsequent crumbling of pellets, extrudates or expandates obtained smaller particles) a (rumen-unstable) mixture of at least one solid particulate feed component with at least one rumen-unstable ingredient added to the mixture, selected from at least one conjugated linoleic acid (CLA) in pure form or as a component of a natural or synthetic mixture of substances, wherein in particular individual, several or all of the mixture components which are to be mixed with the rumen-unstable ingredient for pelleting do not in themselves show any or insufficient rumen-stabilising effect in the mixture to be pelletised.In CLA-containing mixtures, those mixture components which would cause rumen stabilization of the CLA in non-pelletized CLA mixtures are not included or are only included in ineffective amounts. Examples of this include the stabilizing additives contained in the rumen-stabilized CLA commercial product Lutrell®<, such as in particular soybean oil, gypsum or silica, which are not included in the feed mixture to be pelleted according to the invention or are only included in non-stabilizing amounts. The proportion of added rumen-unstable ingredient is according to the invention in the range of 0.1 to 20, 1 to 15, 2 to 10 or 3 to 5 wt. %, based on the total weight of the mixture to be pelleted. The pelleted ruminant feed according to the invention is produced by... a. at least one solid particulate feed ingredient is mixed with the ingredient, b. the mixture thus obtained is optionally conditioned by introducing steam at a temperature of up to 95°C, c. the optionally conditioned mixture is pressed into pellets in a pelletising, extruding or expanding device under the action of pressure and optionally steam at temperatures in the range of 60 to 100°C, and d. the pellets thus obtained are optionally cooled and / or dried and optionally subsequently crumbled.

[0029] The mixture used to produce the feed pellets according to the invention can therefore be described as a "rumen-unstable" mixture, which only acquires "rumen stability" through the pelleting, extrusion or expansion step according to the invention, ie whose rumen instability is thereby reduced.

[0030] It follows that, according to the invention, no feed mixtures are processed into pellets that would already possess sufficient rumen stability prior to pelleting. For example, the invention therefore does not encompass the processing of rumen-protected ingredient mixtures (such as the CLA mixture Lutrell® or mixtures of comparable composition) into pelleted ruminant feed.

[0031] According to the invention, the CLA is selected from a) cis / trans-9,11-linoleic acid, i.e. C18:2 cis-9, trans-11 and C18:2 trans-9,cis 11 b) cis / trans-8,10-linoleic acid; i.e. C18:2 cis-8, trans-10 and C18:2 trans-8,cis 10 c) cis / trans-11,13-linoleic acid; i.e. C18:2 cis-11, trans-13 and C18:2 trans-11,cis 13 d) cis / trans-10,12-linoleic acid; ie C18:2 cis-10, trans-12 and C18:2 trans-10,cis 12 e) cis / cis-9,11-linoleic acid, f) trans / trans-9,11-linoleic acid, g) cis / cis-8,10-linoleic acid h) trans / trans-8,10-linoleic acid i) cis / cis-11,13-linoleic acid j) trans / trans-11,13-linoleic acid k) cis / cis-10,12-linoleic acid l) trans / trans-10,12-linoleic acid; and m) mixtures of at least two of the above-mentioned compounds.

[0032] In particular, the CLA is selected under a) 9-cis,11-trans-linoleic acid (C18:2 cis-9, trans-11); b) 10-trans,12-cis-linoleic acid (C18:2 trans-10, cis 12); and c) mixtures thereof.

[0033] The invention also relates to a process for producing a pelleted ruminant feed according to the invention, wherein a) at least one solid particulate feed component is mixed with at least one rumen-unstable ingredient as defined above, b) the mixture thus obtained is optionally conditioned, in particular by introducing steam, in particular conditioning with hot steam of up to 95 °C, such as saturated steam, over a sufficient period of time, such as for 20 to 40 or 20 to 120 or 20 to 140 or 20 to 240 seconds, c) the optionally conditioned mixture is pressed into pellets in an extrusion, expansion or pelletizing device, in particular a pelletizing or extrusion device, under the action of pressure and optionally steam at a temperature of 60 to 100 °C; and d) the pellets thus obtained are optionally cooled and / or subsequently dried and optionally subsequently crumbled.

[0034] The pelleted mixture obtained has an improved rumen stability of the ingredient (i.e. a reduced rumen instability of the ingredient up to preferably essentially complete rumen protection) compared to the non-pelletized mixture.

[0035] Further described herein, but not within the scope of the claimed invention, is a method for altering the milkfat concentration in milk produced by a lactating ruminant, comprising administering to the lactating ruminant an effective amount of a pelleted ruminant feed as described above. The lactating ruminant is selected from cows, goats, and sheep. d) Further details d1) Production of feed pellets

[0036] To produce the pelleted feed compositions, the rumen-unstable ingredient(s) to be formulated according to the invention are mixed, optionally together with other additives, with conventional animal feed ingredients (such as milk-yielding feed). The ingredient proportion is selected so that it is, for example, in the range of 0.1 to 20, 1 to 15, 2 to 10 or 3 to 5 wt.%. The feed is then pelleted using a suitable pellet press. For this purpose, the feed mixture is usually conditioned by introducing steam (with up to 95°C hot steam, e.g. saturated steam), e.g. for 20 to 1240 seconds. The temperature increase during the pelleting process can be influenced by the amount of saturated steam added. However, this steam conditioning can also be omitted.

[0037] Pelletizing is typically carried out using commercially available pellet presses (e.g., from Bühler GmbH), which may be configured as ring-die presses. The preconditioned material mixture is pressed through a ring die (gap width between roller and die, e.g., 1 mm) by means of rollers. Depending on the die, pellets with a diameter of approximately 2 to 12 mm can be produced. For dairy cattle feed, for example, dies with a bore diameter of 4 to 5 mm and a hole length of 40 to 50 mm are used. The highest process temperature occurs when the mixture is pressed through the die. Temperatures in the range of approximately 60 to 100°C can be reached here.

[0038] The hot material exiting the die outlet is continuously cut off by a knife, and the resulting pellets are immediately cooled in a cooler and, if necessary, further dried. The residual moisture content can range from approximately 1 to 15 wt.% based on the total pellet weight after cooling and, if necessary, drying.

[0039] Alternatively, pelleting can be replaced by extrusion or expansion (e.g., using feed expanders from Kahl, Germany) (see, for example, "Feed Manufacturing Technology IV" Ed: mcEllhiney, Kansas State Univ.; Pub. by American Feed Industry Assoc., 1994; Arlington, VA. Therein: Extrusion Cooking Systems, Bob Hauck et al., pp. 131-139; Wilson et al. 1998, Journal of Poultry Science, 77 (Suppl. 1): 41). The products from these processes can also be recrumbled. d2) Application

[0040] Typical feed ingredients that can be used to produce pellets according to the invention include feed materials of plant or animal origin according to FMV (Feed Regulation), such as cereal products, wheat feed meal, wheat bran; extraction meal, pomace, molasses chips, fish meal, meat-bone meals; and / or mineral feed materials according to FMV, such as carbonates, phosphates, sulfates, propionates.Also suitable are cereals such as wheat, rye, barley, oats, maize, millet or triticale; cereal products (by-products of milling) such as bran, semolina bran, wheat semolina bran, feed flour or secondary flour; by-products from oil production (extraction meal, expeller, cake); by-products from sugar production (molasses, dried pulp, feed sugar, pulp, potato starch, maize gluten, wheat gluten); by-products of the fermentation industry, brewer's spent grains, yeast, malt germ, stillage; as well as animal and other feedstuffs such as blood meal, fish meal, press juice, potato protein.

[0041] In particular, mention should be made of wheat, grain maize, barley, oats; soybeans; cereal flours such as wheat or maize flour, soybean meal, beet molasses pulp, wheat semolina bran, wheat flour, maize gluten feed, soybean meal, rapeseed meal, brewer's grains, grain distillers, beet molasses, oat bran; sugars such as glucose and sugar alcohols, furthermore protein-containing components such as soy concentrate, fish meal, glutens such as maize or wheat gluten, oils and fats, and nutraceuticals such as free amino acids, their salts, vitamins (such as A, D, E) and trace elements (such as Cu as CuSO 4 ), mineral components such as calcium carbonate, sodium chloride; phosphates, and optionally processing aids such as lubricants, inert fillers and the like, and optionally preservatives.

[0042] Typical milk performance feed compositions contain, for example, maize, wheat, barley, oats, rye, citrus pulp, soybean meal, rapeseed meal, soybean hulls, palm kernel expeller, DDGS, maize gluten feed, sugar beet pulp, wheat semolina bran, wheat bran, linseed, molasses, lime, salt, vitamin and trace element premix.

[0043] The invention will now be explained in more detail with reference to the following embodiments. Experimental part Example 1: Investigation of the Milk fat reduction in dairy cattle feeding with different pelleted CLA feed formulations a) Pelletizing

[0044] Pellet press: Manufacturer: Simon Heessen, Type V3-30C. Capacity: 600-650 kg / h. Die: Bore diameter: 5 mm. Bore length: 45 mm. Gap width: 1 mm. Pelletizing temperature: 80°C. Conditioning: Steam T = 135 and 145°C (depending on pressure). b) CLA formulations used (1) Lutalin ®< : (CLA-containing commercial product)

[0045] Table: Composition of 5 different batches fatty acid 1 2 3 4 5 Palmitic acid % 1< 4 4 5,3 6,4 6,5 Stearic acid % 3,6 3,6 3,6 3,5 3,6 oleic acid % 28,4 28,4 26,5 23 22 Linoleic acid % 1,5 1,6 2,3 1,7 1,8 CLA trans-10, cis-12, omega-6 % 29,9 29,9 29,6 30,4 31,0 1< GC Area-% CLA cis-9, trans-11 % 29,9 29,9 29,6 30,4 31,0 CLA total % 59,8 59,8 59,2 60,8 62,0 CLA trans, trans % 0,4 0,4 0,4 0,6 0,4 Peroxide value, Ph.Eur. (B 057) ppm 17,9 6,3 7,1 13,5 11,9 Acid number (ISO 660-96) mg KOH / g 14,0 14,1 14,0 14,9 12,9 Water (ISO 4317) % 0,050 0,077 0,049 0,013 0,184 Methanol (NF EN 14110) ppm 40,0 70,0 40,0 10,0 40,0 Total fatty acids 97,7 97,8 97,3 96,0 96,3 (2) Lutrell ®< :

[0046] Lutrell consists of 34% lutalin, as well as the additives 15% silica, 5% gypsum and 46% hydrogenated soybean oil, the latter in particular fulfilling the rumen-stabilizing function.

[0047] The composition (fatty acid profile) of Lutrell is shown in the following table fatty acid Portion (% total product) C14:0 0,12 % C15:0 0,00 % C16:0 7,94 % C16:1 0,04 % C17:0 0,00 % C18:0 41,05 % C18:1, c9 8,18 % C18:2, c9, c12 0,67 % C20:0 0,38 % C18:3 0,00 % CLA cis-9, trans-11 9,39 % CLA trans-10, cis-12, 9,45 % C22:0 0,36 % C24:0 0,12 % Other fatty acids 2,42 % Other substances 20,00 % (inorganic carrier) Σ 100 % (3) Lutalin plus Silafett

[0048] Fatty acid composition Silafett = hardened (hydrogenated) soybean oil C12:0 0-0,5% C14:0 0-1 % C16:0 7-14% C18:0 85-93% C18:1 0-3% C18:2 0-0.5% Trans fatty acids: max. 1% b) Experimental scheme

[0049] Four test variants are tested (see the following table). Variant 1 (T1) is the control (K) and contains no CLA, variant 2 (T2) contains the CLA formulation "Lutrell", variant 3 (T3) contains the CLA formulation "Lutalin" and variant 4 (T4) contains the CLA formulation "Lutalin plus Silafett". T1 T2 T3 T4 Soy-corn mix 1000 1)< 950 983,3 950,0 Lutrell 0 50 0,0 0,0 Lutalin 0 16,7 16,7 Lutalin plus Silafett 0 0,0 33,3 1000 1000 1000 1000 1) Information in grams

[0050] Feed: 1 kg supplement made from 25% soybean meal and 75% maize meal, pre-pelletized

[0051] The CLA preparations are mixed into a milk performance feed (25 wt% soybean meal; 75 wt% corn meal) and pelleted.

[0052] The experimental design corresponds to a 4 x 4 Latin square (see table below). Pure CLA is dosed in equal amounts of 16.7g per cow per day. Group 1 (n=5) Group 2 (n=5) Group 3 (n=5) Group 4 (n=5) Period 1 K Lutalin Lutrell Lutalin + Silafett Period 2 Lutalin K Lutalin + Silafett Lutrell Period 3 Lutalin + Silafett Lutrell K Lutalin Period 4 Lutrell Lutalin + Silafett Lutalin K

[0053] Allocation to results table (section f) below): K= Control = T1, Lutrell = T2, Lutalin = T3 and Lutalin plus Silafatten = T4.

[0054] Each period lasts 21 days. This means that after an adaptation period of 14 days, a measurement period of 7 days follows. All four experimental diets are based on the same basic components. c) Animals

[0055] Each treatment will be tested with 20 cows according to the above experimental scheme. A further 4 cows will be allocated to the experimental groups as reserve animals. The experimental cows will be in the middle lactation phase at the start of the experiment. The average milk yield at the start of the experiment will be approximately 32 kg FECM per day. The experimental cows (German Holstein) will be selected from the herd of the experimental station (herd yield approximately 11,000 kg milk, 3.9% fat, 3.4% protein) and bred according to the following criteria: Lactation number (1 and higher) Milk fat content (based on the two previous MLPs) Lactation day and milk yield

[0056] divided into four groups with the aim of setting comparable group average values. d) Feeding

[0057] All cows are fed the same total mixed ration (TMR), which is given once a day to ad libitum The TMR consists of maize silage, grass silage, hay, and approximately 45% concentrate. The target TMR values ​​are shown in the table below. This TMR ensures that the cows are adequately supplied with 33 kg of milk, assuming approximately 21 kg of DM are consumed. nXP g / kg T XP g / kg T XL g / kg T XF g / kg T NEL MJ / kg T Total 155 155 30 180 6,9

[0058] Content of the TMR in crude nutrients according to Weender analysis (nXP=usable crude protein, XP=crude protein, XL=crude fat, XF=crude fiber and NEL=net energy lactation.

[0059] Pelleted premixes of the three CLA formulations are first prepared with a milk-performance feed. These premixes are then manually added to the TMR in the feed troughs in appropriate proportions (equivalent to 1 kg per animal per day), with the goal of achieving a concentration of 16.7 g of each pure substance per 20 kg of TMR. The control group receives the same concentrate in the same amount without CLA. e) Sampling and measurement parameters feed

[0060] A sample is taken daily from the TMR for dry matter determination, which is combined into a single sample per measurement period (7 days). This sample is used to determine the crude nutrient content using the Weender analysis and the energy content using the Hohenheim feed value test. cows

[0061] For each cow, TMR intake, milk yield, and live weight are recorded daily (individually). During the 7-day measurement period, milk samples are taken three times as aliquots from the evening and morning milkings and analyzed for fat, total protein (Nx6.38), lactose, urea, and somatic cell levels by the Baden-Württemberg Milk Testing Association. An additional milk analysis is performed in the middle of the first and second week of each trial period (preparation phase). In addition, a collective milk sample covering the 7-day measurement period is taken and frozen for each cow and treatment to allow for a fatty acid analysis of the milk fat, if necessary. f) Test results

[0062] Table: Effect of Lutalin and Lutrell in pelleted feed on milk fat reduction in dairy cows fed 5 g trans-10 / cis-12 CLA Treatment Milk kg Fat kg Protein kg Protein % Fat % MFD T1 = Control Mean 34,4 1,310 1,153 3,40 3,86 0% SD 6,56 0,232 0,170 0,38 0,58 T2 = Lutrell Mean 34,6 1,190 1,146 3,36 3,52 - 9 % SD 7,60 0,220 0,200 0,38 0,59 T3 = Lutalin Mean 35,1 1,218 1,156 3,34 3,52 - 9 % SD 7,27 0,218 0,183 0,37 0,54 T4 = Lutalin plus / Sillafett - 9 % Mean 34,7 1,205 1,151 3,36 3,50 SD 6,56 0,224 0,167 0,37 0,46 MEAN: mean SD: standard deviation

[0063] Surprisingly, treatments T3 and T4 resulted in the same milk fat reduction as treatment T2. This was despite the active ingredient CLA in T3 and T4 being mixed into the milk performance feed in a non-rumen-protected form. Pelleting therefore protected the active ingredient CLA from T3 and T4 in the rumen to the same extent as known from Lutrell and also demonstrated by T2. Example 2: Investigation into the rumen stability of different pelleted CLA feed formulations

[0064] Stability in the rumen is a key prerequisite for milk fat reduction using CLA-containing feeds. For this purpose, the feeds tested in Example 1 (75% corn plus 25% soybean meal) were mixed with 1.66% Lutalin or 5% Lutrell and pelleted or extruded. The resulting pellets and extrudates were then incubated in vitro for 4, 12, and 24 hours. After the specified times, the incubation was stopped, the contents were transferred into a container, and freeze-dried. The CLA content of this freeze-dried sample was then determined. a) Pelletizing and extrusion

[0065] Preparation of the meal: Corn and soybean meal were weighed together, ground, and mixed in a horizontal mixer. Lutalin and Lutrell, respectively, were added and mixed. The mixing time was 15 minutes. These two mixtures were then divided into two parts: 80 kg were pelleted and 30 kg were extruded.

[0066] Pellet mill: Simon Heesen with a horizontal die, 3.3 x 35 mm, nominal capacity 300 kg / hr.

[0067] Extruder: Werner & Pfleiderer 37, die 2 x Ø 3.9 mm, nominal capacity up to 50 kg / hr.

[0068] Pelleting: Pelleting temperatures of 67-68.5 °C were achieved.

[0069] Extrusion: An extrusion temperature of 85°C was reached, and approximately 14% water was added. A residual moisture content of 10 to 12% was found in the final product. b) CLA formulations used

[0070] (1) Lutalin ®< : (CLA-containing commercial product. Composition see Example 1.) (2) Lutrell ®< Pure: CLA-containing commercial product. Composition see Example 1 c) Experimental scheme

[0071] Four test variants were tested (see the following table). Variant 1 (T1) A pellet containing the CLA formulation "Lutalin" Variant 2 (T2) A pellet containing the CLA formulation "Lutrell" Variant 3 (T3) An extrudate containing the CLA formulation "Lutalin" Variant 4 (T4) An extrudate containing the CLA formulation "Lutrell" T1 T2 T3 T4 Soy-corn mix 983,3* 950 983,3 950,0 Lutrell 50 0,0 50 Lutalin 16,7 16,7 0 sum 1000 1000 1000 1000 * Information in grams

[0072] Feed: 1 kg supplement made from 25% soybean meal and 75% maize meal.

[0073] An analysis immediately following processing showed that the active ingredient CLA survived pelleting very well but that extrusion led to approximately 40% loss of activity. d) Incubation: Determination of in vitro degradation products in the Hohenheim feed value test (in vitro test)

[0074] The Hohenheim Feed Value Test (HFT) is an in vitro method for estimating the energy value of ruminant feed. In an adapted version, the HFT can also be used to test active ingredients for their rumen stability by stopping incubation after defined times and analyzing the remaining active ingredient in the incubation solution.

[0075] The Hohenheim Feed Value Test (HFT) is carried out according to the method specifications of the VDLUFA (Method Book Vol. III, Chapter 25.1).

[0076] The incubation time depends on the specific research question and is appropriately between 4 and 24 hours for the degradation of CLA formulations. The in vitro system is stable for up to 48 hours, but deviations from a physiological fermentation process are to be expected beyond this time. 500 mg of each of the test variants T1 to T4 (corresponding to approximately 5 mg CLA or approximately 3 mg for the extruded variants) were weighed. The incubation times were 4, 12, and 24 hours. A triple determination (3 incubation vessels per test variant T1 to T4) was performed for each incubation time.

[0077] After the appropriate time, incubation was stopped by cooling in ice water. The rumen fluid buffer mixture was quantitatively transferred into glass vessels suitable for subsequent freeze-drying. The drying vessels were weighed empty so that the residual weight after drying could be determined and used for subsequent quantitative analysis. A Gamma 1-20 from Christ, 37507 Osterode, Germany, was used as the freeze-drying system. e) Method of analysis: Modification of AM / 00887 / 01

[0078] Due to the special matrix and the low CLA content, the preparation described in the analytical method (see Chapter 7.4.2) had to be modified as follows. The changes made are shown in italics.

[0079] The sample is weighed or placed into a 250 ml Erlenmeyer flask as described in 7.4.2, then the specified amounts of BHT, sodium ascorbate and water are added.

[0080] Then add a spatula tip of the enzyme pronase and place the Erlenmeyer flask in an ultrasonic bath heated to 60 °C for 15 minutes.

[0081] After adding the amounts of ethanol and acetic acid described in 7.4.2, the sample is placed in the ultrasonic bath heated to 60 °C for another 15 minutes.

[0082] After cooling to room temperature, the extraction solution cyclohexane / ethyl acetate 80:20 (v,v) is added. However, only 50 ml of the extraction solution is used instead of the usual 100 ml.

[0083] The mixture is then stirred on a magnetic stirrer for 30 minutes as described in 7.4.2. 70 ml of saturated sodium chloride solution is added and stirred for another 10 minutes. The stirrer is then turned off and phase separation is allowed to occur.

[0084] 3 ml of the organic phase are taken, the solvents cyclohexane and ethyl acetate are removed in a nitrogen stream and the residue is taken up in 2 ml of the extraction solution cyclohexane / ethyl acetate 80:20 (v,v) (concentration).

[0085] A portion of the resulting sample solution is filtered directly into an HPLC vial using a 0.45 µm disposable filter.

[0086] The injection volume is 20 µl instead of the usual 10 µl. f) Test results Retention (%) CLA total (as methyl ester and as free fatty acid)

[0087] Sample weight, mg mg retention Retention in % of starting weight Retention time, h Retention time, h 0 4 12 24 0 4 12 24 Pellet Lutalin 5,1 1,49 0,32 0,09 100% 29% 6% 2% Pellet Lutrell 5,3 2,09 0,71 0,14 100% 39% 13% 3% Extrudate Lutalin 3,05 2,76 1,22 0,16 100% 91% 40% 5% Extrudate Lutrell 2,95 1,10 0,54 0,54 100% 37% 18% 18%

[0088] The identical milk fat reduction of pelleted Lutalin compared to pelleted Lutrell, which was surprisingly observed in Example 1, could be reproduced in this in vitro experiment insofar as the measured CLA retention rates were identical in both pelleted variants T1 and T2. Thus, this test can be used to reliably assess at least those variants that are based on the same feed and differ only in terms of physical treatment. Surprisingly, the extruded variants T3 after 4 and 12 hours, and the T4 after 12 and 24 hours, each showed significantly higher retention rates than the pelleted variants (see Figure 1). Figur 1 ).

[0089] It can therefore be concluded that Lutalin (non-rumen-stabilized CLA) in extruded form also has at least equivalent rumen stability to a comparable extrudate made from Lutrell (rumen-stabilized CLA through stabilizing additives).

Claims

1. A pelleted ruminant feed for dairy cattle, comprising in uncoated, pelleted form a mixture of at least one solid particulate feed component with at least one rumen-unstable constituent which has been added to the mixture, selected from at least one conjugated linoleic acid (CLA), wherein the proportion of added constituent is in the range of from 0.1 to 20% by weight, based on the total weight of the mixture to be pelleted, and the pelleted ruminant feed is prepared in that a. at least one solid particulate feed component is mixed with the constituent, b. the resulting mixture is optionally conditioned by introduction of steam at up to 95°C, c. the optionally conditioned mixture is compressed in a pelleting, extruding or expanding device with the action of pressure and optionally steam at temperatures in the range of from 60 to 100°C to give pellets, and d. the resulting pellets are optionally cooled and / or subjected to a final drying process and optionally subsequently crumbled.

2. The feed according to claim 1, wherein the constituent is a CLA, selected from among 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 abovementioned compounds.

3. The pelleted ruminant feed according to claim 2, wherein the at least one solid particulate feed component is selected from cereals, cereal derivatives, such as by-products of milling; by-products of oil production, such as extracted meals, expellers and cake; by-products of the production of sugar, such as molasses, dry beet chips, feeding sugar, pulp, potato starch, maize gluten and wheat gluten; by-products of the fermentation industry, such as brewer's grains, yeast, malt culms and stillage; and feedstuffs of animal and other origin, such as blood meal, fish meal, pressed must and potato protein.

4. The feed according to claim 1 or 3, wherein the constituent is a CLA, selected from among a) 10-trans,12-cis-linoleic acid; and b) mixtures of 9-cis,11-trans-linoleic acid and 10-trans,12-cis-linoleic acid.

5. The feed according to claim 4, wherein the CLA is selected from among mixtures of 9-cis,11-trans-linoleic acid and 10-trans,12-cis-linoleic acid.

6. The feed according to claim 5, wherein the CLA mixture is added in the form of one of the following fatty acid compositions: Fatty acid12345Palmitic acid%445.36.46.5Stearic acid%3.63.63.63.53.6Oleic acid%28.428.426.52322Linoleic acid%1.51.62.31.71.8CLA trans-10, cis-12, omega-6%29.929.929.630.431.0CLA cis-9,trans-11%29.929.929.630.431.0CLA total%59.859.859.260.862.0CLA trans, trans%0.40.40.40.60.4Peroxide number, Ph. Eur. (B 057)ppm17.96.37.113.511.9Acid number (ISO 660-96)mg KOH / g14.014.114.014.912.9Water (ISO 4317)%0.0500.0770.0490.0130.184Methanol (NF EN 14110)ppm40.070.040.010.040.0Total fatty acids97.797.897.396.096.3 where the % values are gas chromatography area% values.

7. A process for the preparation of a pelleted ruminant feed according to any of the preceding claims, wherein a. at least one solid particulate feed component is mixed with at least one constituent as defined in any of claims 1 to 6, b. the resulting mixture is optionally conditioned by introduction of steam at up to 95°C, c. the optionally conditioned mixture is compressed in a pelleting, extruding or expanding device with the action of pressure and optionally steam at temperatures in the range of from 60 to 100°C to give pellets, and d. the resulting pellets are optionally cooled and / or subjected to a final drying process and optionally subsequently crumbled.

8. The process according to claim 7 for the preparation of a rumen-protected ruminant feed, wherein in step a. a rumen-unstable mixture of the constituent with the at least one feed component is obtained and wherein the pelleted mixture obtained in step b. has an improved rumen stability of the constituent as compared to the unpelleted mixture.

9. The feed or process according to any of the preceding claims, wherein the mixture to be pelleted is pelleted in a pelleting press designed as an annular die press by pressing the pre-conditioned substance mixture through the annular die by means of rollers.

10. The feed or process according to any of the preceding claims, wherein the annular die has a bore diameter of 4 to 5 mm and a slot length of 40 to 50 mm.

11. The feed according to any of claims 1 to 6, wherein the proportion of added constituent is in the range of from 1 to 20% by weight, in particular 3 to 20% by weight, based on the total weight of the mixture to be pelleted.