NZ205249A - Ruminant feed additives containing zinc complexes and amino acids - Google Patents

Abstract

1. Particulate feed additive for ruminants on the basis of water-insoluble zinc complexes of one or more essential amino acids, characterized in that it consists of a mixture of the water-insoluble zinc complexes of one or more essential amino acids with a substantially rumen-proof matrix.

Description

New Zealand Paient Spedficaiion for Paient Number £05249 2 05249 Priority Date(s): J At. V' Complete Specification Filed: Class: »tti« 4PR Publication Date: . ..rr.... P.O. Journal, No: .. ma me.

AMENDED under Section of tfee Patents Act 1953 frc/ ASSIStAtff "COHMISSION£R OF PATENTS NO DBA' S NEW ZEALAND PATENTS ACT, 1953 No.: Date: COMPLETE SPECIFICATION PARTICULATE FEED ADDITIVE FOR RUMINANTS I/We, LOHMANN TIERERNAHRUNG GmbH, Neufelder Street, 24-28 2190 Cuxhaven, Federal Republic of Germany, <=» ks=>t^i CZjzz?*vpz*s=ayl^. hereby declare the invention for which / we pray that a patent may be granted to me/us, and the method by which it is to be performed, to be particularly described in and by the following statement:- - 1 - (followed by page la) 2052 ia rARTICULATE FEED ADDITIVE FOR RUMINAN-T-S The invention relates to a particulate feed additive for ruminants, which is intended to improve the supply of essential amino acids 5 to ruminants.

Considerable efforts have been made in the past to improve the supply of nutrients, such as e.g. proteins 03?" free essential amino acids to ruminants. In the case of oral administration 10 the main problem is that for these substances to be effectively available to the animal, .they must pass substantially undamaged through the rumen or paunch. For this purpose, for example, amino acids are protected by chemical treatment *15 or by enveloping with a rumen-proof coating.

However, this leads to the further problem that, although the amino acids must be protected in the rumen, this protection must not prevent the resorption of the amino acids in the following 20 digestive tract. It has not hitherto proved possible to achieve the objective of a 100% protection in the rumen and a 100% resorption in the following digestive tract. It has in fact been found that in the case of amino acids 25 protected in known manner, either the protection in the rumen is inadequate, or the resorption in the following digestive tract failed to meet expectations. This was more particularly shown in in vivo tests, in which the content of 30 essential amino acids in the blood serum was tested as a function of the administration of protected amino acids. Reference is made in this 2 205&49 connection to an article by Dr. K. Ktither in "Rinderwelt", no. 1, 198$, pp. 3 to 8 and to Degussa-Inf ormationsdienst ftlr die Mischfutter-industrie 1/1979, pp. 1 to 5, (both available on request), which 5 reveal the latest stage of development, particularly in connection with what the Experts consider to be the most important amino acid for ruminants and in particular dairy cows with a high milk yield methionine. In connection with the numerous 10 proposals relating to methods for protecting biologically active substances in the rumens of ruminants, reference is made e.g. to U.S.

Patent 3959493, DE-OS 30 13 000, U.S. Patent 3 656 864, British Patent 1 137 214, Norwegian 15 Patent 120 058 and the various publications referred to therein. (The German and Norwegian patents are available on request.) In view of the aforementioned inadequacies of the prior art, the present invention is based on the problem of supplying a feed additive for 20 ruminants, which can not only be produced in a very simple manner, but, when fed to ruminants, leads to a better combination of passage through the rumen and resorption in the following digestive tract of essential amino acids than the 25 hitherto known feed additives, so as to improve the actual availability of the essential amino acids.

To solve this problem, a particulate feed additive for ruminants of the type characterized 30 in the claims, as well as for a process for the production thereof are proposed.

As stated hereinbefore, at present methionine is considered to be the most important essential 14F£B©86"v) 205249 amino acid for ruminants, i.e. it is looked upon as the first limiting amino acid. Thus, the invention is mainly described hereinafter by means of a feed additive according to the 5 invention, which improves the supply of methionine to ruminants.

Zinc-Ot-amino acid complex salts and .particularly zinc-methionine complex salts with a molar zinc : amino acid ratio of 1 : 1 10 are known (cf. DE-AS 24 36 946 (available on request), U.S. Patents 3 463 858, 3 941 818 and 4 021 569). These are water-soluble complex salts, whose positively charged cation consists of a zinc-oc-amino acid complex, whilst the anion can be selected at 15 random (cf. e.g. the structural formulas given in DE-AS 24 36 946 (available on request), and U.S. Patents 3 948 818, and 4 021 569). The aforementioned publications are essentially based on the fact that these zinc compounds are particularly readily accessible 20 zinc sources for the human and animal body. It is further mentioned in passing, that these compounds are also suitable substances for supplying animals with essential amino acids. These publications make no mention of the special 25 problems occurring in ruminants. This is not surprising, because the aforementioned complex salts are water-soluble compounds, which dissolve in the rumen when administered to ruminants and consequently do not reach the following parts of ^ ' 30 the digestive tract and can consequently not provide an additional supply of essential amino acids to ruminants. 203249 4 It is also mentioned in passing in DE-AS 24 36 946 (column 2, lines 36 to 40) (available on request) , U.S. Patent 3. 941 848 (column 3, lines 55 to 59 and U.S. Patent 4 021 569 (column 4, lines 10 to 15) that when reacting water-soluble zinc salts with methionine in the alkaline range, a water-in-soluble complex of methionine and zinc is formed with a molar zinc : methionine ratio of 1 : 2. However, no details are given on the character-10 istics of this complex. The only teaching which is given is that the formation of a water-insoluble complex should be avoided.

According to the invention, it has now surprisingly been found that the water-insoluble "15 zinc complexes formed during the reaction of essential amino acids with water-soluble zinc salts at a suitable pH-value are relatively rumen-proof, i.e. they pass through the rumen when administered to ruminants. With falling pH-20 value, and at the latest after hydrochlori-c acid hydrolysis in the abomasum at pH 2-4, the compounds according to the invention are again decomposed into their components and are available for resorption in the following parts 25 of the digestive tract. The availability in the digestive tract portions following the rumen and consequently the resorption which can be followed by means of the blood serum values can be further considerably increased if the rumen-30 tightness is improved, in that the particulate, water-insoluble zinc - amino acid complexes are mixed or enveloped with a prior art, rumen-proof matrix. 205249 The feed additive according to the invention is produced by dissolving methionine at*' ambient temperature in water by adding acid, e.g. hydrochloric acid. Accompanied by stirring, a 5 zinc salt, e.g. zinc sulphate (anhydrous or hydrated) is then added to the solution and the salt rapidly dissolves. The methionine and zinc salt are used in a molar ratio of 2 : 1. The clear solution obtained is then set to a pH-value 10 of 7.5, accompanied by thorough mixing, which preferably takes place by the dropwise addition of a lye. After setting the pH-value to 7.5, stirring is continued for a time until the precipitate is separated, washed with water and 15 dried. The production of water-insoluble zinc complexes of histidine, phenyl alanine and valine takes place in the same way. It is also mentioned in this connection that the precipitates obtained sometimes do not consist solely of the 20 water-insoluble zinc complexes and amino acids and sometimes considerable quantities of co-precipitated zinc hydroxide can be obtained.

According to the preferred embodiment of the invention, the particulate, water-insoluble 25 zinc-methionine complex produced in the above manner is mixed or enveloped with a rumen-proof matrix. This takes place in accordance with per se known processes, e.g. by spray crystallization (cf. German Patent 24 51 510 - available on request). 30 Other suitable methods are described in the publications referred to in connection with rumen-proof coatings. Particular reference is made to DE-OS 30 13 000 (available on request) and Chemie, Ingenieur und Technik 51, 205249 1979, no. 11, pp. 996 to 998 and 45, 1973, no. 23, p. 1339. It is obviously also possible to mix or envelop mixtures of zinc complexes of two or more essential amino acids with a rumen-proof matrix.

The materials known from the prior art for this purpose are suitable for the rumen-proof matrix for mixing with the zinc complexes according to the invention or for enveloping the zinc complexes according to the invention. These 10 include e.g. triglycerides, saturated fatty acids and their salts, mixtures of monoglycerides, diglycerides and triglycerides and mixtures of glycerol tristearate with liquid, unsaturated higher fatty acids and combinations of the 15 aforementioned protective materials. Reference is made to the aforementioned specifications and the publications quoted therein and additionally reference is made to DE-OS 28 38 27 8, DE-OS 28 38 298 and DE-OS 28 38 309 (all available on request) . 20 Based on the total weight of the feed additive according to the invention, the rumen-proof matrix quantity is 0 to 90% and preferably 45 to 65%. However, in general every attempt should be made to keep the rumen-proof matrix proportion as low as possible. 25 The particle size of the feed additive according to the invention and consisting of zinc complex and rumen-proof matrix is in the range 0.3 to 1.0 mill, preference being given to a range of approximately 0.3 to 0.6 mm. This relatively 30 small particle size represents a not inconsiderable advantage compared with the known rumen-passing. feed additives with a rumen-proof matrix produced with a particle size of roughly 1.0 mm and higher. Although this leads to an increase ; ^ 2 052 in the surface area it simultaneously shortens the passage time through the psalterium. Mixing advantages are also obtained, because these finer particles are roughly of the same size 5 as the constituents of meal-like (unpelletized) feed mixtures and consequently have no demixing or separating tendency.

An important disadvantage of the prior art rumen-traversing feed additives is that they 10 cannot be pelletized, because the chemical or ' physical protection is damaged during pelletizing due to thermal and mechanical influences. This limits the use of the known rumen-traversing feed additives, because nowadays mixed feeds 15 are preferably produced in pelletized form.

However, as opposed to this, the.feed additive according to the invention can be pelletized without significantly impairing the protective action in the rumen. This is due to the 20 considerable thermal stability of the zinc complexes according to the invention and to their lack of sensitivity to water during the conditioning with hot steam which takes place directly prior to pelletizing, so that the slight reduction 25 in the protective action of the matrix due to pelletizing is unimportant.

The dosing of the feed additive according to the invention is obviously dependent on the requirement of the particular animal for the 30 different essential amino acids. Thus, for example, in effectivity tests on cows, such a feed additive quantity has proved suitable as corresponds to an additional administration of 205249 8 1 to 30 g of methionine daily and preferably approximately 15g of methionine daily. It is to be expected due to the higher amount of methionine reaching the blood serum that, 5 compared with the known products, the daily administration of the feed additive according to the invention can be reduced. Particular importance is attached to the additional oral administration of methionine to sheep, because 10 sulphur-containing amino acids have a considerable influence on wool formation. For a sheep, with a body weight of approximately 50 kg, the dosing of the feed additive according to the invention corresponds to a quantity representing an 15 additional administration of approximately 5 g of methionine.

The dosing of feed additives according to the invention containing amino acids other than methionine, can be determined by animal tests, 20 as are described e.g. in the following examples.

Example 1 89.4 g (0.6 mol) of methionine were dissolved in 1500 ml of water at ambient tempera-25 ture by adding 200.ml of 1 n hydrochloric acid, subsequently and accompanied by stirring, 86.2 g (0.3 mol) of zinc sulphate (ZnSO^ x 7H^0) were added to this solution and the salt rapidly dissolved. The clear solution obtained was 30 brought to a pH of 7.5 by the dropwise addition of 45% caustic potash solution, accompanied by vigorous stirring. At the end of addition, stirring continued for 10 minutes, the precipitate *4 Feb $93$ 2 052 was separated, washed with water (2 x 300 ml) and dried at 100°C.

The yield of zinc-methionine complex in the ratio 1 : 2 (empirical formula: 5 C^QH^N^O^S^Zn) was 104.1 g (= 95.9% of theory). The zinc "and methionine contents of the product were investigated by routine analytical processes. The following values.were determined: 18.5% zinc (theory: 18.15%) and 78.5% methionine (theory: 10 81.85%).

Example 2 29.8 g (0.2 mol) of methionine were dissolved in 500 ml of water at ambient temperature, accompanied by the addition of 40 ml of 2 n 15 hydrochloric acid. Subsequently 8.14 g (0.1 mol) of zinc oxide were added to the solution, which was also completely dissolved, accompanied by the addition of dilute hydrochloric acid. The mixture was then set to a pH-value of 7.5 by the 20 dropwise addition of a 45% potassium hydroxide solution. The precipitated complex was separated, washed with 600 ml of water and dried at 110°C. The yield was 33.0 g (= 91.2% of theory) and the analysis gave 18.4% zinc and 77.6% methionine. 25 The zinc-methionine complex obtained in Examples 1 and 2 can be further purified by additional, conventional purification processes, although this is not necessary for the purposes / i of the invention. Due to ! the volume and quantity ratios chosen in Examples 1 and 2, formation of the precipitate already commenced before. reaching a pH-value of 7.5. 2 05249 Example 3 The zinc-methionine complex produced in Example 1 was tested in a feeding test ori cows. - A proven method for determining the amino 5 acid requirement in animals vis an increasing administration of the particular amino acid with blood samples taken in parallel. The level in, the serum remains constant for as long as the amino acid requirement" is not covered. As soon 10 as the requirement is covered (and possibly another amino acid has a limiting action), there is a marked rise in the content in the blood serum. In this connection reference is made to Degussa-Informationsdienst flir die Mischfutter-15 industrie 1/1979, pp. 1 to 5, where the above method is described in detail and the facts are shown graphically on p. 2.

For the comparative testing of the protective action and availability of the protected 20 amino acids in ruminants, it is advisable to initially administer a quantity which is beyond the animal's requirement, so that the expected serum content is always on the rising curve (see graph in the aforementioned literature reference) 25 In this way it is possible to test different rumen-traversing feed additives with a clearly defined methionine content and compare them e.g. with the serum values obtained by abomasum infusion. This procedure has the advantage that 30 with the methionine quantity reaching the blood serum it is simultaneously possible to determine the protection in the rumen and the availability in the following parts of the digestive tract. 205249 11 As the passage time through the psalterium and also the resorption rate can differ when using different methionine forms, the tests performed used frequent small oral administrations, 5 whereas infusions took place continuously.

The methionine forms and quantities used in the tests performed are given in the following table, together with the blood serum values measured. The individual cows were in each case 10 treated for one week. Oral administrations took place in six, uniformly distributed and identical portions daily between 6 a.m. and 6.30 p.m.

T ab 1 e 1 Methionine content in 15 serum* pg/ml Methionine form Cow 1 Cow 2 Without methionine administration 4.2 4.8 60g (in 6 doses) of 20 orally administered methionine complex (l : 2) daily, corresponding to approx. 47g of DL-methionine 5.8 5.8 10 g of infused DL-methionine in the abomasum daily 11.9 18.3 * Each value resulted from the mean of three analyses of three blood sampling times. The test 30 animals were lactating cows.

The results given in the above Table shows that the oral administration of the zinc-methionine complex according to the invention significantly 2 05249 # 12 increases the methionine content in the blood serum, which is not the case when using water-soluble methionine. However, after the infusion of methionine, the blood serum contents are 5 nowhere near reached.

Example 4 The tests described in Example 3 were repeated with the zinc-methionine complex according to the invention and the zinc complex was 10 mixed with a matrix of saturated triglycerides. This mixture was prepared according to the aforementioned spray crystallization process, in which the hot melt prepared from the zinc complex and the triglycerides was sprayed under 15 pressure against a cold air stream. Calculated as methionine, the zinc complex content was 30%. For comparison purposes, normal methionine was mixed in the same way with the aforementioned matrix and once again the methionine content was 20 30%.

The test results obtained are given in the following Table 2. The serum samples were taken at 2 p.m. on the last day of the particular treatment period, i.e. after treatment for one 25 week. # 205249 13 Table 2 Methionine form 5 (amount of daily administration) 150 g of normally protected methionine (30%) = 45 g 10 methionine 7.5 g of infused DL-methionine in the abomasum without methionine administration g of infused DL-me thionine in the abomasum Methionine content in serum* pg/ml Cow 1 4.1 6.3 2.8 150 g of protected methionine according to the invention (30%) = 45 g of methionine 9.4 Cow 2 4.2 14.1 3.1 12.5 .6 18.8 * Each value results from the mean of two analy-25 ses. The test animals were once again lactating cows.

The above results show that the zinc-methionine complex according to the invention which alone only increased the blood serum values to a 30 limited extent, led to an extraordinary rise in effectively resorbed methionine quantity in conjunction with the matrix of saturated triglycerides. The above results also clearly show that the use of the protective matrix of saturated r' 2 05249 14 triglycerides alone only has a comparatively limited action.

The protective matrix used had the following fatty acid jcomposition (saturated 5 fatty acids): C16 : 20.6% C18 : 53.8% C2Q : 13.8% C22 : 7.8% others: 4.0% 100 % 2 052 EXAMPLE 5 The zinc-methionine complex produced according to Example 2 was mixed as described in Example 4 with a matrix of di- and triglycerides and processed according to the spray crystallization process. The resultant product was tested as described in Example 3 in a feeding test on cows, in which for comparison five additional products were tested which are commerically available or produced according to the prior art and which have been described or proved to have rumen-bypass properties. All products were administered in such a dosage that the cows got 45g methionine daily (distributed on six portions of 7.5 g each).

Two cows with a medium-low milk performance (on an average 18 I and 33 l/day, respectively) were used as test animals. The cows were always treated in the same way. The two cows showed a differently distinct reaction with regard to the methionine administrations (cow 2 showed a stronger reaction). These individual differences are known. Between the used product types good comparability was obtained with regard to the ranking of the amounts of methionine "arriving" in the blood serum.

The single values of the analysis carried out (sampled each at II a.m. and 2 p.m.) showed pretty high variations. Therefore, at least four single analysis per product and cow were carried out. As in Example 3, the individual cows were in each case treated for one week.

Besides the product according to the invention designated with D the following further products were investigated for comparative purposes: 2 05249 16 Product A: Hydroxymethy l-methionine-ca Icium salt (Degussa-Mepron) Product B: Methionine-hydroxy-analogue—ca leium_ sa 11 Product C: Methionine enveloped with a matrix of di- and tr i g I ycer i des and an emulsifier.

Product D: Methionine protected according to the invention, which also was enveloped with a matrix of di- and tr i g I yceri des and an emulsifier (as Product C) Product E: Methionine which was enveloped with a matrix consisting of substantially saturated fatty acids.

Product F: Methionine which was also enveloped with a matrix of substantially saturated fatty acids.

A The Products A and B are commerically available. Instead of methionine Product A contains methionine in which the amino group is substituted by a hydroxymethy I group. Product B is a compound correspond i ng to methionine in which, however, the amino group is replaced by a hydroxy group. In both cases these compounds are converted by the animal in vivo to methionine. Products E and F are two similar types which only differ slightly in the fatty acid composition of the matrix.

The test results obtained are given in the following table 3. 205249 NOW AMENDED Week Methionine Product Methionine content in Serum ug/ml Cow 1 ' Cow 2 1 without methionine administration 2. 7 2 Product A 3. 3 3 without methionine administration 3. 6 4 Product B 3. 8 Product C / . 2 6 without methionine administration 3. 9 7 Product D (Invention) 8; 1 8 without methionine administration 3. 8 9 Product E / 4. 4 without methionine administration 3. 1 11, Product F 4. 12 without? methionine administration 3. 6, The average methionine level in the blood/ serum of cow 1 during _/ / the period without methionine adminisarati< / ion was significantly lowgr than the level of cow 2. The higher value in Week 8 thout methionine administration /obtained for cow 2 may be caused by an aftereffect of the methionine protected according to the invention since the firsts/single value in this week was significantly enhanced and was 5.6 yug/ml.

The results given in the above table show clearly that of the six forms of the tested/protected methionine the form according to the invention provided by far the best results and accordingly was far superior to the other products. As in Example 4, the above results a/lso demonstrate that the use of a protective matrix alone has a comparatively minor effect.

Claims (8)

TAsaiv.lnded 17 205249 Table 3 Week Methionine Product Methionine content in Serum ,ug/ml Cow 1 Cow 2

1 without methionine administration

2. 7 3. 8 2 Product A 3. 3 4. 5 3 without methionine administration 3. 6 3. 4 4 Product B 3. 8 7. 0 5 Product C 5. 2 8. 1 6 without methionine administration 3. 9 4. 2 7 Product D (Inventii on) 8. 1 15. 6 8 without methionine administration 3. 8 5. 6 9 Product E 4. 4 7. 7 10 without methionine administration 3. 1 4. 1 11, Product F 4. 5 7. 5 12 without methionine administration

"'3. 6 5. 2 The average methionine level in the blood serum of cow 1 during \ the period without methionine administration was significantly lower than the level of cow 2. The higher value in Week 8 without methionine administration obtained for cow 2 may be caused by an aftereffect of the methionine protected according to the invention since the first sinqle value in this week was significantly enhanced and was 8.75^ug/ml. The results given in the above table show clearly that of the six forms of the tested protected methionine the form according to the invention provided by far the best results and accordingly was far superior to the other products. As in Example 4, the above results also demonstrate that the use of a protective matrix alone has a comparatively minor effect. 15 MAY1986Z) 305249 WHAT^f/WE CLAIM igj Particulate feed additive for ruminants on the basis of water-insoluble zinc complexes of one or more essential amino acids, characterized in that it consist^of a mixture of the water-insoluble zinc complexes of one or more essential amino acids with a substantially 2- Feed additive according to claim 1, characterized in that the rumen-proof matrix forms a coating enveloping the zinc complex particles. Feed additive according to claim 1 or 2, characterized in that it comprises water-insoluble zinc complexes of methionine, histidine, phenyl alanine and/or valine.

4. Feed additive according to claim. 3, characterized in that the water-insoluble zinc complex is [h3c-s-ch2-ch2-ch-coo] 2 Zn nh2

5. A particulate feed additive according to claim 1 and substantially as herein described with reference to any embodiment disclosed in the examples. 205249 19

6. Process for producing a particulate feed additive for ruminants characterized in that an essential amino acid is dissolved in water, accompanied by the addition of acid, subsequently and accompanied by stirring, a water-soluble zinc salt is added in a quantity making the molar ratio of amino acid to zinc salt 2:1, the clear solution obtained is brought to a pH-value of 7.5 by adding lye and accompanied by vigorous stirring, the stirring is continued for a short time when addition is at an end and then the precipitate is separated, washed with water and dried, the precipitate obtained is mixed in per se known manner with a known substantially rumen-proof matrix and/or the precipitate particles are coated with this matrix.

7. Process according to claim 6, characterized in that methionine is used as the amino acid and [H3C-S-CH2-CH2-CH_C00] 2 Zn NH2 is obtained as a precipitate.

8. A process for producing a particulate feed additive according to claim 6 and substantially as herein described with reference to any embodiment disclosed in the examples.