USE OF A FEED COMPOSITION FOR REDUCING METHANE EMISSION IN RUMINANTS, AND/OR TO IMPROVE RUMINANT PERFORMANCE

Abstract

The present invention relates to the field of reduction of methane emission in ruminants. Particularly, it relates to the use of a feed composition or a feed additive comprising at least one fatty acid containing at least 5 carbon atoms and at least one organic molecule substituted at any position with at least one nitrooxy group, for reducing the production of methane emanating from the digestive activities of ruminants, and/or to improve the ruminant performance.

Claims

1. Use of a feed composition or feed additive comprising at least one fatty acid containing at least 5 carbon atoms, and at least one organic molecule substituted at any position with at least one nitrooxy group, or a salt thereof as defined by formula (I), ##STR00035## wherein Y is an organic molecule of the following composition: C.sub.aH.sub.bO.sub.dN.sub.eS.sub.g, wherein a is comprised between 1 and 25, b is comprised between 2 and 51, d is comprised between 0 and 8, e is comprised between 0 and 5, g is comprised between 0 and 3, for reducing the formation of methane emanating from the digestive activities of ruminants, and/or for improving ruminant performance.

2. The use according to claim 1, wherein a is comprised between 1 and 10, b is comprised between 2 and 21, d is comprised between 0 and 6, e is comprised between 0 and 3, g is comprised between 0 and 1.

3. The use according to claim 1, wherein the organic molecule of formula (I) is a compound of formula (II), ##STR00036## wherein n is comprised between 0 and 12, preferably comprised between 0 and 6 and, wherein, if n0, the carbon chain is a linear, a cyclic, or branched aliphatic carbon chain which may be non substituted or substituted with up to 3 hydroxyl, alkoxy-, amino-, alkylamino-, dialkylamino- or nitrooxy groups, or an alkenyl, or alkynyl carbon chain mono- or polyunsaturated and in any isomeric form, R4 is independently, hydrogen or a saturated straight, cyclic or branched chain of an alkyl or alkenyl group containing 1 to 12, preferably 1 to 6 carbon atoms, X is hydrogen, R5, R5N, OR5, OCOR5, NR5R6, ONO2, COOR5, CONR5R6, NHSO2R5, or SO2NHR5, R5 and R6 are independently, hydrogen, C1-C12 straight, branched or cyclic alkyl chain, non substituted or substituted with up to 3 hydroxyl-, alkoxy-, amino-, alkylamino-, dialkylamino- or nitrooxy groups, alkenyl, or alkynyl carbon chain which may be mono or polyunsaturated, and in any isomeric form.

4. The use according to claim 1, wherein the organic molecule of formula (I), or a salt thereof is selected from the group consisting of 3-nitrooxypropanol, ethyl-3-nitrooxy propionate, methyl-3-nitrooxy propionate, and 3-nitrooxy propionic acid, preferably 3-nitrooxypropanol.

5. The use according to claim 1, wherein the fatty acid containing at least 5 carbon atoms has the general formula C.sub.nH.sub.2n+1COOH, wherein n is an integer selected in the range of 4 to 27, preferably in the range of 6 to 20, more preferably in the range of 8 to 15, most preferably in the range of 9 to 13.

6. The use according to claim 5, wherein the fatty acid containing at least 5 carbon atoms is lauric acid.

7. The use according to claim 1, wherein the ruminant animal is selected from the group consisting of: cattle, goats, sheep, giraffes, American Bison, European bison, yaks, water buffalo, deer, camels, alpacas, llamas, wildebeest, antelope, pronghorn, and nilgai.

8. The use according to claim 1, wherein the methane production in ruminants is reduced by at least 10% when measured in metabolic chambers and based on the treatment with the at least one organic molecule substituted at any position.

9. The use according to claim 1, wherein the amount of the at least one organic molecule as defined in formula (I) administered to the ruminant animal is from 1 mg to 10 g per kg feed, and the amount of fatty acid administered to the ruminant animal is from 0.1 to 20 g per Kg of feed.

10. The use according to claim 1, wherein the mol ratio of the organic molecule as defined in formula (I) to the at least one fatty acid containing at least 5 carbon atoms is comprised between 0.00001 and 1, preferably between 0.0001 and 0.5, more preferably between 0.0025 and 0.1, most preferably between 0.004 to 0.01.

11. A feed composition or feed additive comprising at least one fatty acid containing at least 5 carbon atoms and an organic molecule of formula (I) according to claim 1.

12. The feed composition or feed additive according to claim 11 wherein the mol ratio of the organic molecule of formula (I) to the at least one fatty acid is comprised between 0.00001 and 1, preferably between 0.0001 and 0.5, more preferably between 0.0025 and 0.1, most preferably between 0.004 to 0.01.

13. A method for reducing the production of methane emanating from the digestive activities of ruminants and/or for improving ruminant animal performance comprising orally administering to the animal a sufficient amount of a feed composition or feed additive comprising at least one fatty acid containing at least 5 carbon atoms, and at least one organic molecule substituted at any position with at least one nitrooxy group, or a salt thereof as defined by formula (I), ##STR00037## wherein Y is an organic molecule of the following composition: C.sub.aH.sub.bO.sub.dN.sub.eS.sub.g, wherein a is comprised between 1 and 25, b is comprised between 2 and 51, d is comprised between 0 and 8, e is comprised between 0 and 5, g is comprised between 0 and 3,

14. A method according to claim 13, wherein the ruminant animal is selected from the group consisting of: cattle, goats, sheep, giraffes, American Bison, European bison, yaks, water buffalo, deer, camels, alpacas, llamas, wildebeest, antelope, pronghorn, and nilgai.

15. A method according to claim 13, wherein the compound of formula (I) is 3-nitrooxypropanol and the fatty acid is lauric acid.

Description

EXAMPLES

[0086] In Vitro Test for Methane Production:

[0087] A modified version of the Hohenheim Forage value Test (HFT) was used for testing the effect of specific compounds on the rumen functions mimicked by this in-vitro system.

[0088] Principle:

[0089] Feed is gadded into a syringe with a composition of rumen liquor and an appropriate mixture of buffers. The solution is incubated at 39 C. After 8 hours the quantity (and composition) of gas phase produced is measured and put into a formula for conversion.

[0090] Reagents:

[0091] Mass Element Solution: [0092] 6.2 g potassium dihydrogen phosphate (KH.sub.2PO.sub.4) [0093] 0.6 g magnesium sulfate heptahydrate (MgSO.sub.4*7H.sub.2O) [0094] 9 ml concentrated phosphoric acid (1 mol/l) [0095] dissolved in distilled water to 1 l (pH about 1.6)

[0096] Buffer Solution: [0097] 35.0 g sodium hydrogen carbonate (NaHCO.sub.3) [0098] 4.0 g ammonium hydrogen carbonate ((NH.sub.4)HCO.sub.3) [0099] dissolved in distilled water to 1 l

[0100] Trace Element Solution:

[0101] 13.2 g calcium chloride dihydrate (CaCl.sub.2*2H.sub.2O) [0102] 10.0 g manganese(II) chloride tetrahydrate (MnCl.sub.2*4H.sub.2O) [0103] 1.0 g cobalt(II) chloride hexahydrate (CoCl.sub.2*6H.sub.2O) [0104] 8.0 g iron(III) chloride (FeCl.sub.3*6H.sub.2O) [0105] dissolved in distilled water to 100 ml

[0106] Sodium Salt Solution: [0107] 100 mg sodium salt [0108] dissolved in distilled water to 100 ml

[0109] Reduction Solution: [0110] first 3 ml sodium hydroxide (c=1 mol/l), then 427.5 mg sodium sulfide hydrate (Na.sub.2S*H.sub.2O) are added to 71.25 ml H.sub.2O [0111] solution must be prepared shortly before it is added to the medium solution

[0112] Procedure:

[0113] Sample Weighing:

[0114] The feed stuff is sieved to 1 mmusually TMR (44% concentrate, 6% hay, 37% maize silage and 13% grass silage)and weighed exactly into 64 syringes. 4 of these syringes are the substrate controls, which display the gas production without the effect of the tested compounds. 4 other syringes are positive control, in which bromoethane sulfonate has been added to 0.1 mM. When needed, 4 syringes contain a carrier control (if the test compounds need a carrier). The remaining syringes contain the test substances, by groups of 4 syringes.

[0115] Preparation of the Medium Solution: [0116] The components are mixed in a Woulff bottle in following order: [0117] 711 ml water [0118] 0.18 ml trace element solution [0119] 355.5 ml buffer solution [0120] 355.5 ml mass element solution [0121] The completed solution is warmed up to 39 C. followed by the addition of 1.83 ml sodium salt solution and the addition of reduction solution at 36 C. The rumen liquor is added, when the indicator turns colourless.

[0122] Extraction of the Rumen Liquor:

[0123] 750 ml of rumen liquor are added to approximately 1,400 ml of medium solution under continued agitation and CO.sub.2-gassing.

[0124] Filling the Syringes, Incubation and Determining Gas Volumes and VFA Values:

[0125] The diluted rumen fluid (24 ml) is added to the glass syringe. The syringes are then incubated for 8 hours at 39 C. under gentle agitation. After 8 hours, the volume of gas produced is measured, and the percentage of methane in the gas phase is determined by gas chromatography.

[0126] Results

[0127] The food fermented was artificial TMR (44% concentrate, 6% hay, 37% maize silage and 13% grass silage). Lauric acid from Sigma Aldrich was used at a concentration of 1000 M. 3-Nitrooxy-propanol (3-NOP) was used at two different concentrations, 2.5 M and 5 M.

[0128] The results are presented in the following Table 2 and 3. Clear synergistic effects were obtained for the combination of 3-NOP and Lauric acid in view of the methane reduction as well as on the acetate/propionate ratio, which translates into additional performance benefit for the animal. The results depicted below result from the average of three experiments with either Lauric acid, 3-NOP, or a combination of both. Ace.=acetate; Prop.=propionate

TABLE-US-00002 TABLE 2 Effect on methane production 3- Lauric Total Reduction Expected NOP acid methane vs. control reduction* Synergy Treatment [M] [M] [mmol over 8 h] [%] 1 (Con- 0.267 trol) 2 (Ref) 2.5 0.244 0.023 3 (Ref) 5 0.097 0.17 4 (Ref) 1000 0.255 0.012 5 (Inv) 2.5 1000 0.125 0.142 0.232 46 6 (Inv) 5 1000 0.022 0.245 0.085 74 *Expected reduction based on an additive effect vs. control (Found value expected value)/expected value * 100%

TABLE-US-00003 TABLE 2 Effect on volatile fatty acids (VFA) profile, i.e. acetate/propionate ratio 3- Lauric Ace./ Reduction Expected NOP acid Prop. vs. control reduction* Synergy Treatment [M] [M] [Mole Ratio] [%] 1 (Control) 2.26 2 (Ref) 2.5 2.09 0.17 3 (Ref) 5 1.63 0.63 4 (Ref) 1000 1.98 0.28 5 (Inv) 2.5 1000 1.63 0.63 1.81 9.9 6 (Inv) 5 1000 1.29 0.97 1.35 4.4 *Expected reduction based on an additive effect vs. control (Found value expected value)/expected value * 100%