Animal feeds containing specific glycolipids

Abstract

Use of a sophorolipid in the preparation of a feed supplement for the improvement of feed efficiency in an animal. A feed supplement comprising a sophorolipid and lysolecithin mix is also provided for improving the feed conversion rate of an animal.

Claims

1. A supplement composition consisting essentially of: (i) at least one sophorolipid having the following general formulas (1) or (2): ##STR00008## wherein R represents hydrogen or an acyl group having 1 to 4 carbon atoms; R.sup.1 represents hydrogen or an acetyl group; R.sup.2 represents hydrogen or alkyl group having 1 to 9 carbon atoms; R.sup.3 represents a saturated or unsaturated hydrocarbon group containing 6 to 18 carbon atoms; and X represents a functional group selected from the group consisting of an aldehyde, a hydroxyl, a carboxylic acid, an acyl group, an amide and a monosaccharide having a glycosidic linkage; and (ii) an amorphous silicon dioxide, wherein the supplement composition is a powder and wherein the at least one sophorolipid and the amorphous silicon dioxide in the supplement composition are present in a 1:1 ratio, and wherein the supplement composition is suitable for an animal feed.

2. The supplement composition as claimed in claim 1 wherein the at least one sophorolipid has the general formula (1) or (2) below: ##STR00009## wherein R.sup.1 represents hydrogen or an acetyl group; R.sup.2 represents hydrogen or alkyl group having 1 to 9 carbon atoms; and R.sup.3 represents a saturated or unsaturated hydrocarbon group containing 15 to 18 carbon atoms.

3. The supplement composition as claimed in claim 1 wherein R.sup.2 is a hydrogen or a methyl group.

4. The supplement composition as claimed in claim 1, wherein R.sup.3 is a hydrocarbon group containing 16 to 18 carbon atoms.

5. The supplement composition as claimed in claim 1 wherein R.sup.3 has one site that is unsaturated (C═C bond).

6. An ingredient, premix or supplement composition, said composition consisting essentially of: (i) at least one sophorolipid the following general formulas (1) or (2): ##STR00010## wherein R represents hydrogen or an acyl group having 1 to 4 carbon atoms; R.sup.1 represents hydrogen or an acetyl group; R.sup.2 represents hydrogen or alkyl group having 1 to 9 carbon atoms; R.sup.3 represents a saturated or unsaturated hydrocarbon group containing 6 to 18 carbon atoms; and X represents a functional group selected from the group consisting of an aldehyde, a hydroxyl, a carboxylic acid, an acyl group, an amide and a monosaccharide having a glycosidic linkage; (ii) and an amorphous silicon dioxide, wherein the composition is a powder, and wherein the at least one sophorolipid and the amorphous silicon dioxide in the composition are present in a ratio of 1:1 and wherein the dosage of said sophorolipid in said composition is 0.01 wt. % or higher, based on the total weight of said ingredient, premix or supplement; and (iii) at least one of vitamins, trace elements, minerals and organic acids, and wherein the composition is suitable for an animal feed.

7. The composition as claimed in claim 6, wherein the at least one sophorolipid includes at least one sophorolipid of the following general formulas (1) or (2): ##STR00011## wherein R.sup.1 represents hydrogen or an acetyl group; R.sup.2 represents hydrogen or alkyl group having 1 to 9 carbon atoms; and R.sup.3 represents a saturated or unsaturated hydrocarbon group containing 15 to 18 carbon atoms; and wherein the dosage of said sophorolipid in said animal feed composition is 0.01 wt. % or higher, based on the total weight of said animal feed composition, wherein said animal feed composition further comprises vitamins, trace elements, minerals and organic acids.

8. The composition as claimed in claim 6, wherein the composition is suitable for feeding mammals, birds and fish.

9. The supplement composition as claimed in claim 1, wherein the supplement composition is suitable for feeding mammals, birds and fish.

10. A method for increasing the feed conversion rate in an animal comprising feeding a mammal, bird or fish with the supplement composition according to claim 9.

11. A method for increasing feed conversion rate in an animal comprising feeding a mammal, bird or fish with the composition according to claim 8.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) Embodiments of the invention shall now be described, by way of example only, with reference to the accompanying drawings in which:

(2) FIG. 1 is a bar chart illustrating Feed Conversion Ratio for birds treated with a feed additive of Example 1 of the present invention and a control group;

(3) FIG. 2 is a graph of the total Feed Conversion Ratio for control and treated groups of birds of Example 1;

(4) FIG. 3 is a graph of the Feed Conversion Ratio for control and treated groups of birds of Example 1 on the final day (Day 35);

(5) FIG. 4 is a graph of the finisher body weight in kilograms (Day 35) for control and treated groups of birds of Example 1;

(6) FIG. 5 is a graph of the total consumption in kilograms for control and treated groups of birds of Example 1;

(7) FIG. 6 is a graph of percentage dressing for control and treated groups of birds of Example 1;

(8) FIG. 7 is a bar chart illustrating Feed Conversion Ratio at Days 1-7 for birds treated with a feed additive of Example 2 of the present invention and a control group;

(9) FIG. 8 is a bar chart illustrating Feed Conversion Ratio at Days 8-21 for birds treated with a feed additive of Example 2 of the present invention and a control group;

(10) FIG. 9 is a bar chart illustrating Feed Conversion Ratio at Days 22-35 for birds treated with a feed additive of Example 2 of the present invention and a control group; and

(11) FIG. 10 is a bar chart illustrating the total FCR for birds treated with a feed additive of Example 2 of the present invention and a control group of birds.

DETAILED DESCRIPTION

(12) The present invention relates to the use of sophorolipids as an animal feed supplement. A sophorolipid is a surface-active glycolipid that can be synthesized by non-pathogenic yeasts, the main one being Candida bombicola using raw ingredients such as vegetable oils. Sophorolipids reduce surface tension and therefore are used as surfactants in detergents but also possess a number of useful biological activities including antimicrobial, virucidal, anticancer and inmuno-modulatory properties. Sophorolipids have previously been used for the treatment of skin and hair (see, for example, WO 95/34282 and U.S. Pat. No. 5,981,497).

(13) The production of sophorolipids is well documented in the art, see for example, “Production of sophorolipids by the yeast Candida bombicola using simple and low cost fermentative media” Davery and Pakshirajan, Food Research International 42 (2009) 499-504.

(14) Sophorolipids are comprised of a sugar head sophorose, an unusual β-(1,2) disaccharide consisting of two glucose molecules, covalently linked to a long chain hydroxylated fatty acid having a fatty acid chain length of 16-18 carbon atoms. Two types of esterification can occur which have a big effect on the molecule's behaviour, namely (i) acetylation and (ii) lactonization. Acetylation takes place at the C6′ and C6″ positions of the sophorose resulting in a mixture of non-, mono- and di-acetylated molecules. Acidic sophorolipids are not readily soluble in aqueous medium at acidic pH but become soluble at higher pH. However, these sophorolipids show instability at pH higher than 7.0. Lactonization converts the open-chain or acid sophorolipid to a closed-chain sophorolipid (lactonic form), specific for position C4″, see below. The lactonised sophorolipids are stable across a wider pH range.

(15) (i) Acidic Sophorolipid:

(16) ##STR00004##
(ii) Lactonic Sophorolipid.

(17) ##STR00005##

(18) The present invention has surprisingly found that both acidic and lactonic sophorolipids may be added to animal feedstuffs to improve the performance of the animal. This is surprising because sophorolipids, especially the acidic form, would be expected to be destroyed by the stomach acid in the stomach of the animal thereby resulting in minimal, if any, effect on the bodyweight of the animal. Previously, oral administration of sophrolipids has not been considered due to it being readily hydrolyzed in the stomach as well as intestine. Natural sophorolipids containing around 50% of the acidic and lactonic forms may be used as the feed additive. The lactonic sophorolipids are preferred.

Example 1: Trial to Demonstrate Increased Animal Performance for Birds Treated with an Animal Feed Containing a Feed Additive According to an Embodiment of the Invention Comprising a Sophorolipid and Lysolecithin Mix Compared with a Control Group of Birds that Did not Receive the Animal Feed Additive

(19) Birds were fed identical diets over a period of 35 days but the diet of the treated birds was supplemented with 125 g of a sophorolipid/lysolecithin mix per ton of feed that has been cultivated from C. bombicola. 50% of the sophorolipid was in the lactonic form, thus the feed contained 62.5 g per ton of feed. The mix was 75% lysolecithins and 25% sophorolipids at a range of inclusion levels up to 250 ppm. The control and treated groups consisted each of replicates of 10 birds (total of 100 birds for each group). The three phases of the trial were BOOSTER (Day 1-14), STARTER (Day 15-28) and FINISHER (Day 35).

(20) The data obtained for the control group and treated group are provided in the tables below:

(21) I. Control Group.

(22) TABLE-US-00001 Weight (kg) Rep Initial Booster Starter Finisher 1 0.047 0.343 1.360 1.691 2 0.047 0.339 1.261 1.745 3 0.046 0.336 1.206 1.608 4 0.045 0.355 1.226 1.698 5 0.047 0.381 1.322 1.752 6 0.046 0.365 1.238 1.658 7 0.046 0.335 1.220 1.672 8 0.046 0.368 1.327 1.771 9 0.046 0.379 1.326 1.766 10 0.047 0.388 1.336 1.778 0.046 0.359 1.282 1.714 Rep Booster Starter Finisher Total Consumption (kg) 1 0.465 1.437 0.963 2.865 2 0.446 1.463 1.002 2.911 3 0.463 1.387 0.919 2.769 4 0.455 1.434 0.996 2.885 5 0.482 1.461 0.940 2.882 6 0.460 1.409 1.057 2.926 7 0.528 1.440 0.936 2.903 8 0.468 1.523 1.013 3.003 9 0.500 1.540 0.925 2.965 10 0.495 1.476 1.089 3.060 0.476 1.457 0.984 2.917 FCR 1 1.569 1.413 2.905 1.742 2 1.528 1.587 2.068 1.714 3 1.597 1.594 2.286 1.772 4 1.468 1.647 2.108 1.745 5 1.443 1.551 2.187 1.690 6 1.444 1.614 2.511 1.815 7 1.827 1.626 2.070 1.785 8 1.452 1.588 2.280 1.741 9 1.500 1.626 2.100 1.723 10 1.448 1.557 2.464 1.767 1.523 1.578 2.278 1.749 Weight Gain (kg) 1 0.296 1.017 0.332 1.645 2 0.292 0.922 0.485 1.698 3 0.290 0.870 0.402 1.562 4 0.310 0.871 0.473 1.653 5 0.334 0.942 0.430 1.705 6 0.319 0.873 0.421 1.612 7 0.289 0.886 0.452 1.627 8 0.322 0.959 0.444 1.725 9 0.333 0.947 0.441 1.721 10 0.342 0.948 0.442 1.732 0.313 0.923 0.432 1.668 Dressing Rep Dressing (%) Weight 1 80.000 1.400 2 76.744 1.650 3 77.778 1.400 4 78.378 1.450 5 78.947 1.500 6 80.000 1.400 7 75.00 1.350 8 77.143 1.350 9 76.923 1.500 10 75.676 1.400 84.020 1.440
II Treated Group

(23) TABLE-US-00002 Weight (kg) Rep Initial Booster Starter Finisher 1 0.046 0.341 1.305 1.733 2 0.046 0.345 1.197 1.648 3 0.045 0.364 1.290 1.711 4 0.045 0.322 1.215 1.726 5 0.047 0.368 1.227 1.638 6 0.047 0.362 1.294 1.766 7 0.046 0.382 1.262 1.716 8 0.047 0.349 1.219 1.736 9 0.046 0.339 1.237 1.743 10 0.047 0.369 1.241 1.653 0.046 0.354 1.248 1.70 Rep Booster Starter Finisher Total Consumption (kg) 1 0.505 1.506 0.935 2.945 2 0.430 1.398 0.933 2.760 3 0.456 1.506 0.954 2.916 4 0.418 1.446 1.029 2.893 5 0.508 1.413 0.937 2.858 6 0.479 1.460 0.940 2.879 7 0.533 1.451 0.929 2.913 8 0.444 1.456 0.995 2.894 9 0.426 1.428 1.027 2.881 10 0.504 1.392 0.904 2.799 0.470 1.445 0.958 2.874 FCR 1 1.710 1.562 2.183 1.745 2 1.438 1.639 2.070 1.723 3 1.428 1.626 2.263 1.749 4 1.506 1.619 2.014 1.720 5 1.581 1.644 2.279 1.795 6 1.518 1.567 1.994 1.675 7 1.585 1.649 2.046 1.744 8 1.470 1.673 1.922 1.713 9 1.452 1.590 2.030 1.697 10 1.565 1.595 2.193 1.742 1.526 1.615 2.090 1.730 Weight Gain (kg) 1 0.295 0.964 0.428 1.687 2 0.299 0.853 0.451 1.602 3 0.319 0.926 0.422 1.667 4 0.278 0.893 0.511 1.682 5 0.321 0.859 0.411 1.592 6 0.316 0.932 0.472 1.719 7 0.336 0.880 0.454 1.670 8 0.302 0.870 0.518 1.690 9 0.293 0.898 0.506 1.697 10 0.322 0.873 0.412 1.607 0.308 0.895 0.458 1.661 Dressing Rep Dressing (%) Weight 1 82.051 1.600 2 78.378 1.450 3 77.778 1.400 4 77.778 1.400 5 80.000 1.400 6 80.000 1.600 7 77.778 1.400 8 75.676 1.400 9 75.000 1.350 10 77.778 1.400 84.368 1.440

(24) These results are shown graphically in FIGS. 1 to 6 of the accompanying drawings, with the line charts presenting the 10 replicates' results ranked from highest to lowest to demonstrate the consistency of the treated group compared with the control group.

(25) The overall conversion ratio or rate (FCR) was shown to be lower in the treated group than the control group (see FIGS. 1 to 3), most notably during the finisher phase, indicating that the treated group are more efficient converters of their food. The benefit of the feed additive can also be seen in the final bodyweight of the two groups, which was broadly the same, and the lower consumption for the treated group compared to the control group (see FIGS. 4 and 5). The dressing percentage was also higher in the treated group (see FIG. 6).

(26) Modifications to the sophorolipid molecules have also been carried out in the art and such modified sophorolipids are also suitable for use in the animal feed supplement according to the invention. Examples of such modified sophorolipids are given below (i to vii). Their methods of production are described in articles, such as “Enzymatic Synthesis of a Galactopyranose Sophorolipid Fatty Acid Ester” Nunez, Foglia and Ashby, Biotechnology Letters (2003) 25: 1291-1297 and “Enzyme-catalyzed regioselective transesterification of peracylated sophorolipids” Carr and Bisht, Tetrahedron 59 (2003) 7713-7724.

(27) ##STR00006## ##STR00007##

Example 2: Trial to Demonstrate Increased Animal Performance for Birds Treated with an Animal Feed Containing a Feed Additive According to Another Embodiment of the Invention Comprising Sophorolipids Only Compared with a Control Group of Birds that Did not Receive the Animal Feed Additive

(28) Another trial was carried out in a similar manner to Example 1 above with control and treated groups of birds with the birds being fed identical experimental diets over a period of 35 days but the diet of the treated birds was supplemented with a pure sophorolipid (SPL) supplement at a range of inclusion levels up to 250 ppm. The sophorolipid was provided as a powder with 50% sophorolipids liquid with 50% silicon dioxide.

(29) The composition of the Experimental diet is provided below:

(30) TABLE-US-00003 Starter Grower Finisher Ingredients Corn (3 mm) 52.45 7.83 Corn (5 mm) 39.99 18.60 Corn (7 mm) 8.00 40.00 SBM (de-hulled) 39.15 35.55 32.05 Soy Oil 3.59 4.36 5.33 L-Lysine Sulfate 55 0.31 0.21 0.21 DL-Met 98 0.38 L-Met 99 0.32 0.31 Threonine 98 0.10 0.07 0.06 Choline Chloride 50 0.07 0.07 0.08 MCP 1.75 MDCP 1.68 1.60 Limestone (Fine) 1.54 1.25 1.10 Salt 0.30 0.25 0.25 NaHCO.sub.3 0.10 0.19 0.19 Vit Premix-Broiler 0.14 0.14 0.12 Min Premix-Poultry 0.11 0.11 0.11 Total 100.00 100.00 100.00 Calculated values (%) AME.sub.n (kcal/kg) 3,000 3,100 3,200 CP 23.00 21.50 20.00 Lys 1.44 1.29 1.19 Met 0.71 0.64 0.61 Met + Cys 1.08 0.99 0.94 Ca 0.96 0.87 0.81 Available P 0.48 0.43 0.41
Results: Effect of Pure SPLs on Growth Performance in Broilers.

(31) Table 1 below provides the average daily gain (ADG) (g) for the control and treated birds.

(32) TABLE-US-00004 TABLE 1 Days 1-7 Days 7-21 Days 21-35 Overall Control 12.7 34.5 103.2 57.6 SPL (25 ppm) 12.3 39.5 111.0 62.7 SPL (50-ppm) 12.0 35.1 102.0 57.2 SPL (100 ppm) 11.8 35.7 105.3 58.8 SPL (250 ppm) 11.8 35.0 99.8 56.3

(33) Table 2 below provides the feed intake (FI) (g) for the control and treated birds.

(34) TABLE-US-00005 TABLE 2 Days 1-7 Days 7-21 Days 21-35 Overall Control 34.5 84.9 141.3 101.9 SPL (25 ppm) 34.2 92.7 144.9 106.7 SPL (50-ppm) 33.9 78.6 142.8 101.5 SPL (100 ppm) 33.9 80.5 138.6 98.7 SPL (250 ppm) 33.6 80.8 136.6 98.0

(35) Table 3 below provides the feed efficiency (FE) (g) for the control and treated birds.

(36) TABLE-US-00006 TABLE 3 Days 1-7 Days 7-21 Days 21-35 Overall Control 0.37 0.41 0.73 0.57 SPL (25 ppm) 0.36 0.43 0.77 0.59 SPL (50-ppm) 0.35 0.45 0.71 0.57 SPL (100 ppm) 0.35 0.46 0.76 0.60 SPL (250 ppm) 0.35 0.45 0.73 0.58

(37) Examples 1 and 2 above demonstrate that sophorolipids, both alone and in combination with lysolecithin, enhance the feed conversion rate of animals. Example 2 shows that the incorporation of 25-250 ppm pure sophorolipid into an animal feed consistently improves the FCR of the animal compared with the control birds, particularly during the first 8-21 days of the trial, as illustrated in the Table 4 below and shown graphically in FIGS. 7 to 10:

(38) TABLE-US-00007 TABLE 4 Days 1-7 Days 8-21 Days 22-35 Total Control 2.717 2.461 1.369 1.690 25 ppm 2.780 2.347 1.305 1..626 50 ppm 2.825 2.239 1.400 1.666 100 ppm 2.873 2.255 1.316 1.607 250 ppm 2.847 2.309 1.369 1.665

(39) However, the incorporation of a lysolecithin blend and sophorolipid mix (250 ppm) into the animal feed provided better improvement in the FCR relative to the control birds over the entire 35 day trial, as illustrated in Table 5 below and shown graphically in FIG. 1 of the accompanying drawings:

(40) TABLE-US-00008 TABLE 5 Days 1-14 Days 15-28 Days 29-35 Total Control 1.540 1.602 2.153 1.738 Treated 1.526 1.615 2.090 1.730

(41) Thus, this indicates that a lysolecithin/sophorolipid mix provides an improved animal feed supplement. While the inventor does not wish to be bound to the theory, it is believed that the improvement may be due to the different HLB values of the lipids. The HLB value provides an indication of the size of the hydrophilic portion of molecule relative to the molecular mass of the entire molecule. The HLB value of sophorolipids is higher than lysolecithins, thus the overall HLB value of the combination will be pushed upward. As a result, a mixture should improve the emulsification of oil-in water mixtures, as found in the gut of an animal.

(42) Lecithins, particularly lysolecithins or hydrolysed lecithins may therefore be combined with one or more sophorolipids to provide an animal feed supplement according to the present invention. Various types of lecithin or lysolecithin may be used, including blends thereof. Lecithin is a generic term which applies to any complex mixture of lipids containing phosphoric acid, choline, fatty acids, glycerol, glycolipids, triglycerides and phospholipids, including phosphatidycholine, phosphatidylethanolamine, phosphatidylinositol. More preferably, the lecithin blend includes at least one hydrolysed phospholipid wherein one of the fatty acid chains of the phospholipid is hydrolysed and substituted with hydrogen to yield a lysophospholipid.

(43) A preferred lysolecithin for use in the present invention is that sold under the proprietary name of Lipidol, consisting of lecithins and lysolecithins having the following phospholipid profile:

(44) TABLE-US-00009 Phospholipid Weight % 1-Lysophosphatidylcholine 0.6 2-Lysophosphatidylcholine 6.5 Lysophosphatidylinositol 2.1 Lysophosphatidylethanolamine 3.9 Phosphatidylinositol 8.0 Lysophosphatidic acid 1.9 Phosphatidylcholine 3.7 Phosphatidic acid 0.6 N-Acylphosphatidylethanolamine 0.4 Phosphatidylethanolamine 1.7 Other minor phospholipids 3.9 TOTAL PHOSPHOLIPIDS 33.3