GB2599749A - Supplementary feed - Google Patents

Abstract

A supplementary feed, e.g. for livestock, ruminants, pigs, fowls, equines, chickens, dogs or cats, comprises a stoichiometric combination of (i) 1 parts of one or more ribonucleotides, and (ii) 1-10 parts glutamate. The supplementary feed preferably includes 1-5 parts glutamate. The one or more ribonucleotides may comprise disodium inosine-5’-monophosphate (IMP) and/or disodium guanosine-5’-monophosphate (GMP), preferably IMP and GMP in a ratio of 1:1. The supplementary feed may comprise at least 1% by weight of ribonucleotides, and at least 20% by weight of hydrolysed vegetable protein (HVP) containing the glutamate, preferably 2-5% by weight ribonucleotides, and 40-90% by weight HVP containing the glutamate. The glutamate may be provided by yeast or yeast extract. The supplementary feed may include bulking agent and/or anti-caking agent. An animal feed is also claimed comprising up to 1% by weight of the supplementary feed. A method of modifying an animal feed is also claimed, comprising providing an animal feed and combining with up to 1% by weight of the supplementary feed. A method of feeding a non-human animal is also claimed, comprising providing the animal with an animal feed either including or supplemented by up to 1% by weight of the feed supplement.

Description

Supplementary Feed

Introduction

This present invention relates to supplementary feed for animals, to animal feed and to methods of preparing and using the same, especially in the feeding of animals.

Background to the Invention

Production of meat for human consumption is a worldwide industry, and modern agriculture has developed to improve both its quantity and quality. Breeds of animals have been developed that put on weight quickly, reducing the time and cost of meat production. Animal feeds high in protein, energy and various other nutrients have in parallel been developed to further promote speedier and more commercial production methods. Recipes for animal feed are continually evolving in response to the pressure for increased production and in response to problems caused by the drive for greater production efficiencies.

Feed can account for about 60-70% of the cost of meat production, e.g. for pigs. It is, therefore, important to get the formulations and component ratios right at each stage of growth. Industrial farmers generally have targets for weight gain of their animals, and it is generally desirable to provide feed that promotes increased animal growth and weight gain for the whole of the growing period.

Monosodium glutamate (MSG) is a known additive for human food, widely used for its pleasant taste-enhancing properties, attributed to its ability to enhance savoury taste-active compounds. The present invention does not, however, relate to human food.

MSG is, nevertheless, known and permitted for some animal feed subject to approval by the relevant regulatory body. MSG can be made via, among other methods, chemical synthesis, protein hydrolysis (historically the main method of production of MSG) and fermentation.

Waldern and van Dyk, in Journal of Dairy Science, Volume 54, Issue 2, February 1971, Pages 262-265 reported on the "Effect of Monosodium Glutamate in Starter Rations on Feed Consumption and Performance of Early Weaned Calves". They noted that differences in daily gain and feed efficiency from glutamate in the -2 -starter were negligible. They also noted similar results in pigs reported in (i) Klas, R. F., 1964, "Monosodium glutamate in pig creep rations", J. Animal Sci., 23: 598, and (fi) Henson, J. N., Bogdonoff, P. D. and Thrasher, G. W., 1962, "Levels of monosodium glutamate in pig starter preference", J. Animal Sci., 21: 999.

Gate!, F. & Guion, P. (1990) reported on "Effects of monosodium I glutamate on diet palatability and piglet performance during the suckling and weaning periods" in Animal Science, 50(2), 365-372, doi:10.1017/S0003356100004815. It was noted here that during the suckling period, the addition of 5 g MSG per kg diet led to an increase of creep food intake from the 18th day post farrowing. However, no increase in weaning weight was observed subsequent to the intake of the diet with added MSG.

Rezaei, R., Knabe, D. A., Tekwe, C. D., et al. reported "Dietary supplementation with monosodium glutamate is safe and improves growth performance in postweaning pigs" in Amino Acids 44,911-923 (2013). Here, it was noted that feed intake was not affected by dietary supplementation with 0-2% MSG and was 15% lower in pigs supplemented with 4% MSG compared with the 0% MSG group.

The use of and effectiveness of MSG in animal feed is, hence, not clear. Some 20 animals appear not to like its taste and may reject MSG-supplemented feed.

In the agricultural industry it is common practice to feed animals according to a phase feeding system. Phase feeding is the process of adapting an animal's diet according to their age and weight. Within the industry, it is well known that there are problems transitioning animals, e.g. pigs, from one feed phase to the next. In particular, there are difficulties transitioning pigs between the creep and weaner feed phases and between the weaner and grower feed phases.

In particular, in certain animals raised initially on milk, the transfer to solid food, weaning, leads to significant problems. Feed intake in the first few days after weaning is key. Animals often have very low feed intake during the early post-weaning period and research shows that in certain animals, e.g. pigs, this severely limits growth potential, increases temperature and management requirements, intensifies morbidity and mortality, and reduces turnover of facilities and capital. The effect of reduced food -3 -intake at this time is significant -in one study it was estimated that for every 100 g of extra feed per day consumed during the first week post-weaning, body weight increases by 1 to 2 kg at the end of the fourth week post-weaning.

Across the board in animal husbandry, farmed animals often suffer from reduced feed uptake after a change in the feed phase, leading to reduced growth rates.

Suppressed feed intake and growth can lead to increased stress in animals which leads to reduced drinking and increased mortality. Stress leading to reduced drinking of water is an identified cause of mortality in particular in young chickens (broilers and egg producers). Reducing stress can lead to improved drinking and feeding which again in turn reduces animal stress -all resulting in improved weight gain.

In attempts to combat this known problem of reduced feed uptake, one or more sweeteners have been used as feed additives. High amounts may need to be used to be effective, which can be costly. At the same time, sweetener use is restricted to maximum levels for certain animal species. Sweetener use can result in improved animal growth but sweeteners can also lead to some reduction in palatability if excess is added. Overall, sweeteners can give notable benefits with respect to feed uptake and the resulting reduction in animal stress. Alternatives to or supplements to sweeteners are, nevertheless, desirable Ribonucleotides as food additives are known. GB 1 068 446 discloses the use of 5'-ribonucleofides and monosodium glutamate in feeding compositions for livestock and poultry. EP 1 157 616 describes a feeding composition for livestock containing 5'-nucleotides and glutamic acid. WO 2011/000824 relates to the use of 5'-nucleotides, monosodium glutamate and yeast extracts as flavourings for food. WO 2010/060316 describes the development of a meat-flavoured yeast extract comprising hydrolysed vegetable protein and nucleotides. Database WPI & CN 109007369 involves the production of a pig feed comprising ribonucleotides and monosodium glutamate. Lastly, ON 104621362 describes a feed additive for suckling sows and piglets comprising yeast hydrolysate, nucleotides and monosodium glutamate. -4 -

It remains desirable to improve feed uptake in animals generally, and particularly after a change in feed phase (e.g. post weaning) and as a result to improve weight gain and reduce stress-related dehydration and morbidity. Therefore, there remains a need to improve the palatability of animal feed.

An object of the present invention is to provide alternative, preferably improved, methods of feeding animals. Related objects are to provide alternative, preferably improved, animal feeds and/or additives for animal feed. It is particularly preferred to provide improved methods of rearing pigs, ruminants and fowl for meat, and to provide additives for the feeds therefor.

A further object of the invention is to provide improved methods for transitioning animals between feed phases. Related objects of the invention are to provide alternative, preferably improved, feeds to aid the transition of animals between feed phases.

Summary of the Invention

Accordingly, the invention provides a method of feeding a non-human animal by supplementing its diet with a combination of glutamate and 5'-nucleotides. In specific trials this has been shown to improve feed uptake and to lead to notable increase in weight gain during piglet rearing. Dietary supplements, being food stuffs not eaten undiluted but added to animal feed in relatively small amounts, are referred to as supplementary feeds herein.

The invention provides supplementary feeds, animal feeds containing the supplementary feeds, methods of modifying animal feeds and methods of feeding animals, all incorporating the combination of glutamate and 5'-nucleotides of the invention

Detailed Description of the Invention

The invention provides a supplementary feed for a non-human animal, comprising a combination of (i) one or more ribonucleotides, and (ii) glutamate. -5 -

When fed to animals together with, e.g. as a supplement to, animal feed this has been shown to result in improved weight gain and growth. Effects are believed to be associated with one or more of improved feed uptake and a reduction in the impact of feed-related stressors together with an improvement in the gustatory characteristics of the feeding stuff.

The supplementary feed may comprise a stoichiometric combination of (i) 10-1 parts of the one or more ribonucleotides, and (ii) 10 -100 parts glutamate. Synergy is achieved using combinations within these ratios.

Preferably, the stoichiometric combination is of (i) 4 -1 parts of one or more ribonucleotides, and 00 10 -40 parts glutamate. These ratios of the components are preferred as they promote further synergy of flavour enhancement in the supplementary feed.

More preferably, the stoichiometric combination is (i) 3 -1 parts of one or more ribonucleotides, and (ii) 15 -30 parts glutamate. In examples, described in more detail below the ratio was approximately 1: 9-10 of ribonucleotides: glutamate. A particularly preferred range of ratios is ribonucleotide: glutamate of 1: 1-5, more preferably 1: 1.5 -4. Specific examples used ratios of approximately 1: 1.8 and 1: 3.2-3.5 (due to varying glutamate content of the glutamate source).

The ribonucleotides are preferably 5'-ribonucleotides. They are suitably selected from one or more of, or two or more of Guanylic acid (Guanosine Monophosphate; E626), Guanylate salts e.g. Disodium Guanylate (Disodium Guanosine-5'-Monophosphate; GMP; E627), Dipotassium Guanylate (Dipotassium Guanosine-5'-Monophosphate; E628), Calcium Guanylate (Calcium Guanosine-5'-Monophosphate; E629), Inosinic acid (Inosine Monophosphate; E630), lnosinate salts e.g. Disodium lnosinate (Disodium Inosine-&-Monophosphate; IMP; E631), Dipotassium lnosinate 30 (Dipotassium Inosine-&-Monophosphate; E632), mixtures of Guanylates and Inosinates e.g. Calcium-5'-ribonucleotides (E634), Disodium-5'-ribonucleotides (mixtures of IMP and GMP; E635), Calcium lnosinate (Calcium Inosine-&-Monophosphate; E633), Adenylic acid (Adenylate Monophosphate), Adenylate salts -6 - e.g. Disodium Adenylate (Disodium-5'-Adenylate Monophosphate; AMP), Dipotassium Adenylate, Calcium Adenylate, Xanthylic acid (Xanthylate Monophosphate), Xanthylate salts e.g. Disodium Xanthylate (Disodium Xanthosine5'-Monophosphate; X1V1P), Dipotassium Xanthylate (Dipotassium Xanthosine-5'-Monophosphate), Calcium Xanthylate (Calcium Xanthosine-5'-Monophosphate).

Guanylic acid, Guanylate salts, Inosinic acid and lnosinate salts are preferred nucleotides, as are combinations thereof More preferred are IMP and GMP, and particularly preferred is a mixture of IMP and GMP.

The feed according to the invention may thus comprise Disodium Inosine-5'-Monophosphate (IMP) or Disodium Guanosine-5'-Monophosphate (GMP), or mixtures thereof. A mixture of 5 -1 parts by weight IMP and 1 -5 parts by weight GMP is suitable, and a mixture of 3-1 parts by weight IMP and 1 -3 parts by weight GMP is further suitable. 50:50 mixtures of IMP and GMP are commercially and conveniently available, for example "Ribofide® (I + G)" from MCLS Europe and "Agitide® I + G" from Ajinomoto, Japan. These ribonucleotides may also be sourced from, among other things, yeast extracts. In an example below a 1:1 ratio of IMP and GMP was used.

The glutamate may be provided as glutamic acid, glutamate, one or more salts of glutamate, e.g. Magnesium Diglutamate, Calcium Diglutamate, Potassium Glutamate, Sodium Glutamate, Ammonium Glutamate or an alternative. For example in certain embodiments of the invention, the glutamate is provided as vegetable protein rich in glutamates. In other embodiments of the invention, the glutamate may be provided as yeast or yeast extracts.

Hitherto, animal feeds have successfully used sweeteners to try to improve feed uptake. In this invention supplementary feeds may further comprise (iii) a sweetener. Use of sweetener has not been satisfactorily employed to reduce young animal stress.

In the invention, the sweetener is used in part to mask the taste of the combination of glutamate and nucleotide and in part for its contribution to tastiness and attractiveness of the resultant feed. The combination of sweetener plus glutamate plus nucleotides has been found in testing to reduce animal stress and result in improved growth and weight gain. -7 -

Suitable sweeteners in general are those known in the industry for animal use. Typically, a mono-or di-saccharide is used. Examples include thaumafin, sucrose, lactose and dextrose. Artificial sweeteners can also be used, e.g. sodium saccharin, neohesperidin dihydrochalcone; a commercially available example is TastetiteTm (Inroads International Ltd, UK). In an example below dextrose was used, as its monohydrate.

A further optional component is (iv) an anti-caking agent (sometimes referred to as a free flowing agent), especially when the supplementary feed is provided in a dry form; it can be in tablets or pellets, and a preferred form is as a powder. Suitable anti-caking agents include ferricyanide compounds e.g. sodium ferricyanide or potassium ferricyanide, and/or silicon compounds e.g. silicon dioxide, silicic acid, aluminosilicates, kieselguhr, calcium silicate, kaolinific clays, sepiolite, bentonite or vermiculite. An example below uses silicic acid. This agent is included in small amounts, enough to have its effect without adversely affecting overall taste or acceptance of the supplementary feed or feed containing the same. A level of at least 0.3% is common and up to 5% by weight is usually sufficient, more commonly up to 3%, and in particular about 0.5-2%.

The glutamic acid or glutamate component can be provided directly or in various forms or equivalent forms (e.g. as discussed above) or incorporated into one or more other products, and many staple products include glutamates. Suitably it is included as part of a protein-containing ingredient or amino acid-containing ingredient, especially conveniently as a component of a hydrolysable or hydrolysed vegetable protein (also referred to as HVP). HVPs are known as taste and/or flavour enhancers and are widely available commercially in many different forms and from many different suppliers, and at commercially attractive prices.

Generally, the raw materials for HVPs are proteins, broken down into constituent amino acids and used as flavour enhancers, typically soybean or corn. Other vegetable raw materials used in the production of HVP include defatted oil seeds (soy and grapeseed) and maize, wheat, pea, and rice. -8 -

HVP can be produced by boiling one or more of these proteins e.g. soy, corn or wheat in acid, especially hydrochloric acid and then neutralising the solution with alkali, especially sodium hydroxide. The acid breaks down the protein in vegetables into their component amino acids.

For use according to the invention, it is preferred that the HVP contains a source of glutamic acid, e.g. glutamates, at useful levels, enough to combine with the nucleotide components of the feed, and suitably at least 3% by weight, preferably at least 5% by weight, also preferably at least 10% by weight, most preferably at least 15% by weight.

High glutamic acid HVPs may contain up to 40%, more commonly up to 30% glutamic acid by weight. HVPs are staple products. A suitable glutamic acid-containing HVP is available from Exter B.V., based in the Netherlands.

The supplementary feed may therefore comprise: CO the ribonucleotides, being one or more phosphate derivatives of one or more nucleosides; 00 hydrolysed vegetable protein containing the glutamate; (iii) optionally, a sweetener; and (iv) optionally, e.g. a silicon compound as an anti-caking agent / free flowing 20 agent.

In general, the supplementary feed is fairly rich in its key components, as it is added to regular animal feed, and may therefore comprise: (i) at least 1% by weight of one or more ribonucleotides; and (ii) at least 20% by weight of hydrolysed vegetable protein containing the glutamate.

The supplementary feed may contain: (i) at least 2% by weight of one or more ribonucleotides; and OD at least 40% by weight of hydrolysed vegetable protein containing the glutamate. -9 -

In certain embodiments, it may contain up to 5% by weight of one or more ribonucleotides; and up to 90% by weight of hydrolysed vegetable protein containing the glutamate.

If present, the sweetener content is generally at least 20% by weight, preferably at least 30% by weight, more preferably at least 40% by weight.

Preferred embodiments of the invention including the sweetener contain: (i) at least 1.5% by weight of one or more ribonucleotides; (ii) at least 30% by weight of hydrolysed vegetable protein containing the glutamate; and (iii) at least 30% by weight of sweetener.

Further preferred embodiments of the invention including the sweetener contain: (i) at least 2% by weight of one or more ribonucleotides; (ii) at least 40% by weight of hydrolysed vegetable protein containing the glutamate; and (iii) at least 40% by weight of sweetener.

Still further preferred embodiments of the invention contain: (i) at least 2% by weight of a mixture of IMP and GMP; (ii) at least 40% by weight of a hydrolysed vegetable protein containing glutamate; (iii) at least 40% by weight of a sweetener; and (iv) an anti-caking agent It is especially preferred that this product is in the form of a dry powder.

A further optional component is a bulking agent comprising or consisting of calcium carbonate. Use of this component is advantageous when long-term exposure of the supplementary feed risks excessive absorption of water leading to stickiness and handling difficulties. The bulking agent may be present in an amount of generally at least 50% by weight, preferably at least 60% by weight, more preferably at least 70% -10 -by weight. In an embodiment described in more detail below, approximately 75% by weight of the product was calcium carbonate -in the form of limestone flour.

Preferred embodiments of the invention including the bulking agent contain: (i) at least 2% by weight of one or more ribonucleotides; (ii) at least 5% by weight of yeast or yeast extract containing the glutamate; and (iii) at least 50% by weight of bulking agent.

Further preferred embodiments of the invention including the sweetener contain: (i) at least 4% by weight of one or more ribonucleotides; (ii) at least 8% by weight of yeast or yeast extract containing the glutamate; and (iii) at least 60% by weight of bulking agent.

Still further preferred embodiments of the invention contain: (i) at least 8% by weight of a mixture of IMP and GMP; (fi) at least 12% by weight of yeast or yeast extract containing the glutamate; (iii) at least 60% by weight of a bulking agent; and (iv) an anti-caking agent.

It is preferred that the supplementary feed is a complementary feed. In Europe, the definition of a complementary feed is governed by Directive 79/373/EEC. Inter alia, in this legislation 'complementary feed' means a compound feed which has a high content of certain substances but which, by reason of its composition, is sufficient for a daily ration only if used in combination with other feed. Limits are thus generally set for concentration or amount of certain components within the complementary feed based on anticipated dilution in use. The definition was modified by EC Regulation No 767/2009, which requires that complementary feed shall not contain levels of feed additives that are higher than 100 times the relevant fixed maximum content in complete feed or (less relevantly for this invention) five times in case of coccidiostats and histomonostats.

In accordance with required dilution law, therefore, noting however that this can vary in different countries, it is preferred that the supplementary feed is for dilution in use so as to constitute up to 5% by weight resulting after mixing into animal feed (5% supplementary feed, 95% feed), more preferably up to 2% by weight and, at least in Europe, very preferably up to 1%.

In use the supplementary feed is hence added as a supplement, at low levels, to animal feed; animals are then fed with or dosed with the resultant supplemented feed.

Hence, the invention further provides an animal feed comprising the supplementary feed of the invention, with the supplementary feed suitably being at levels as indicated above, suitably at a level of up to 1% by weight; it may be added at up to 0.5% or up to 0.1% by weight or in the range 0.05 -0.1% by weight. In an example below it was added at about 0.1% by weight.

Still further provided by the invention is a method of modifying an animal feed, comprising: (i) providing an animal feed; OD combining (i) with a supplementary feed of the invention suitably at the levels identified immediately above, hence suitably a level of up to 1% by weight.

The invention in addition provides a method of feeding an animal, comprising providing the animal with the supplementary feed of the invention, or providing the animal with an animal feed and supplementing the animal feed at the levels identified immediately above, hence suitably with up to 1% by weight of the supplementary feed of the invention.

Throughout this invention reference to 'animal' is intended to refer to non-human animals and usually this is done specifically by reciting 'non-human'. In all cases, the invention is not intended to relate to humans at all.

In all aspects, the animal is conveniently selected from: livestock, ruminants, fowl, equines, dogs and cats. Herein, equines include horses and donkeys. Herein, fowl include poultry (both broilers and eggs producers).

The animal is preferably selected from pigs, from ruminants, such as cows, sheep and deer, and from fowl such as chicken, geese, turkeys, pheasants and ducks In preferred embodiments the animal is a pig, cow or a chicken, and the examples to -12 -date have used pigs and chickens. In other preferred embodiments the animals are dogs, especially domesticated dogs, i.e. pets. In other preferred embodiments the animals are cats, especially domesticated cats, i.e. pets.

Use of the supplementary feed, as per the example below, has shown it can improve animal rearing.

Examples

Example 1 -Feed Stuff A A complementary feed stuff A (referred to as "Feed stuff A") was prepared as follows, combining umami components for improving feed intake, acceptance and appetite.

Component.k.Neight.Pe ent HVP 47.50 IMP + GMP 2.50 Anhydrous dextrose 48.75 Anti-caking agent 1.25 Notes 1. HVP is hydrolysed vegetable protein: specifically the product "TGLF", supplied by Exter B.V.: the Netherlands. There is some batch to batch variation in the percentage of alutarnic add present in TGLF, with this batch containing approximately 7-8% by weight glutamic acid.

2. IMP ± GMP = equal quantities by weight Disodium Inosine-5'-Monophosphate and Disodium Guanosine-5'-Monophosphate.

3. Anti-caking agent used was 1.00% Precipitated silicic acid E551a plus 0.25% Hydrophobic silicic acid E551a.

The product was a light brown, free-flowing powder with a characteristic odour and savoury salty taste. It had a bulk density of 850 g/L and a tapped bulk density of 1.1 g/L. Analysis showed: moisture -2%, crude protein -10%, crude fibre -0%, crude fat -0.4%, crude ash -13%, calcium -147 mg/kg, sodium -4.4%, phosphorus -0.3%, -13 -lysine -0.2%, methionine -0.1%, magnesium -90 mg/kg. In storage prior to use it was sealed in foil packaging.

Example 2 -Evaluation of feed stuff A in early nursery diets Feed stuff A was evaluated in a piglet feeding trial.

Project Justification and Background:

A. Berry flavouring (from Inroads International Ltd, UK) is currently included in nursery feeds to stimulate intake in weaned pigs.

B. We evaluated feed stuff A (see above) for effect on stimulating intake in pigs.

Objectivity: A. Objective: To determine if the addition of feed stuff A increases intake in weaned pigs.

B. Unknown: The growth performance and morbidity of pigs fed feed stuff A. Experimental Location: 1. Pen Size: 1.5 m x 2.5 m (3.75 m2; -0.25 m2/pig) 2. Floor Type: Concrete slats 3. Feeder Type: 5-hole stainless steel (76.2 cm x 26.7 cm x 2.5 cm) 4. Waterer Type: Gate-mounted stainless-steel cups Experimental Timeline: A. Duration: 22 days Experimental Design: A. Growth Data 1. Design: Randomized complete-block 2. Replication Factor: 18 pens 3. Allotment Factors: visual allotment based on sow parity, body weight, and sex using equal CVs across pens B. Animals: 1 Genetics: DNA 610 E x DNA 241 Fl 2 Number 1,076 pigs 3 Pigs/pen: 14-16 pigs/pen 4 Sex: Split-sexed Age at weaning: -18 -21 d -14 - 6 Body Weights i. Wean weight: 5.66 ± 0.22 kg ii. End Weight: 10.61 ± 0.56 kg Experimental Treatments: A. Control diet without current flavour B. Control diet with current flavour C. As B + feed stuff A 1. 0.45 kg/tonne Ph 1; 0.21 kg/tonne Ph 2 Experimental Procedures: A. Allotment, daily management, live performance, and health measurements followed SOPs as outlined in the company's Monogastric Research Study Manual.

Experimental Measurements: A. Body (pen) weights at allotment/start of experiment (d 0) and weekly after that.

B. Pen feed intakes at each corresponding body weight measurement.

C. Morbidity, mortality, medicinal recordings, & scour scores. Experimental diet formulation, manufacture, and logistics: A. Test ingredient 1. Feed stuff A (recommended inclusion: 0.5-1.0 kg / tonne) B. Experimental feed: 1. Farm manufactured complete experimental Flex Start diets.

2. Berry and feed stuff A flavours were added at the expense of corn.

3. Diets were fed by days as follows: a. Phase 1:8 days b. Phase 2: 7 days c. Common Phase 3: 7 days Results and Discussion: A. Growth performance 1. At day 15 post wean, pigs fed feed stuff A tended to be 0.25 kg heavier (P = 0.100) than those fed no flavouring.

2. Pigs fed feed stuff A were numerically 0.11 kg heavier (P = 0.158) than those fed the current diets on d 22 post-wean.

-15 - 3. ADG was numerically improved 8.3% during week 1 (P = 0.288) and 3.7% (P= 0.151) during week 2 for pigs fed feed stuff A. 4. No other statistical differences were seen in body weights, ADG, ADFI, or FIG. B Health 1. There were no differences in morbidity, mortality, or scour scores between treatments.

-16 -Table 1. Impacts of feeding a savory taste stimulant to early nursery pigs on growth performance.1 Treatments No Berry Flavour (Current) B Berry + Feed stuff A C SEM Trt P-value Flavouring A Body weights, kg Wean, d 0 5.643 5.665 5.661 0.054 0.958 d 8 6.500 6.523 6.604 0.064 0.458 d15 8.128Y 8.255xY 8.378' 0.100 0.100 d22 10.537 10.673 10.786 0.136 0.158 Day 0-22 ADG, kg 0.222 0.227 0.231 0.005 0.116 ADFI, kg 0.308 0.318 0.322 0.009 0.359 F/G, kg/kg 1.40 1.41 1.38 0.02 0.284 Day 0-8 ADG, kg 0.109 0.109 0.118 0.005 0.288 ADFI, kg 0.141 0.150 0.145 0.005 0.689 FIG, kg/kg 1.29 1.38 1.23 Day 9 -/5 ADG, kg 0.231 0.245 0.254 0.009 0.151 ADFI, kg 0.313 0.327 0.322 0.009 0.364 FIG, kg/kg 1.35 1.35 1.28 0.03 0.271 Day 16 -22 ADG, kg 0.345 0.345 0.345 0.009 0.407 ADFI, kg 0.503 0.508 0.522 0.014 0.411 FIG, kg/kg 1.47 1.47 1.52 0.04 0.212 xY Means within a row with different superscripts are different, Pc 0.10.

Data are means of 18 replicates with 14-16 pigs/single-sex pen analysed as a RCBD 5 using the Mixed Procedure of SAS.

-17 -Table 2. Impacts of feeding a savoury taste stimulant to early nursery pigs on nursery health parameters.' Berry Flavour Berry + Feed No Flavouring Trt P-value (Current) stuff A Treatments A Stool looseness2 Day 0 -8 1.11 1.17 1.12 0.483 Day 9 -15 1.52 1.47 1.50 0.858 Day 16 -22 1.83 1.84 1.83 0.843 Day 0 -22 1.45 1.47 1.45 0.889 Medications3 5 7 8 0.824 Removals, n 2 1 2 0.922 Nutritional 2 1 1 0.568 Mortality, n 0 2 1 0.293 1Data are means of 18 replicates with 14-16 pigs/single-sex pen.

2 Stool looseness refer to the average score of the pen. Scours were recorded daily using the following scale: 1 = no looseness, 2 = moderate looseness, 3 = severe looseness / scours 3 Medications are the total number of instances a pig received therapeutic medications regardless of reason.

Example 3 -Feed Stuff B A complementary feed stuff B was prepared as follows, combining umami I5 components for improving feed intake, acceptance and appetite.

Component Weiaht Percent GLY Yeast 15.00 IMP + GMP 7.00 -18 -Limestone flour 75.00 Anti-caking agent 3.00 Notes 1. GLY Yeast is the product "Yeast Extract GLY", supplied by Kohjin Life Sciences Co., Ltd. There is some batch to batch variation in the percentage of MSG present in GLY Yeast, with this batch containing approximately 34.20% by weight MSG.

2. IMP + GMP = equal quantities by weight Disodium I nosine-5'-Monophosphate and Disodium Guanosine-5'-Monophosphate.

3. Limestone flour used was ground calcium carbonate.

ID 4. Anti-caking agent used was precipitated silicon dioxide E551a.

Example 4-Evaluation of feed stuff A in broiler chickens

1. Introduction

Improving feed intake during the brooding phase (0 -10 d post hatch) in broiler chickens is of importance to enhance a good start in early life. In this study we evaluated 2 taste enhancers that may improve feed intake via sensory stimulation. We tested anice, which may attract birds in general by its smell, and feed stuff B of the invention.

2. Materials and methods Experimental design One of the 4 treatments was distributed over floor pens each containing 22 broilers: control, anice, feed stuff A, and anice+feed stuff A. This resulted in 6 replicate pens per treatment. Anice flavour was provided by Greenvalley International (Wageningen, the Netherlands), and products were supplemented at: anice -0.4 kg / tonne, Feed Stuff A -0.3 kg / tonne. The study was only executed in the start phase: 0-10 d post placement.

Diets and validation -19 -Broilers were fed a standard wheat-corn-soybean meal diet without phytase, but with xylanase included and Maxiban included as anti-coccidiostat. Taste enhancers were included at the expense of wheat. Broilers were fed crumbled diets. Diets were formulated and nutrient values calculated based on Schothorst Feed Research (SFR) Feedstuff table (version 24.0125.02). Calculated values of crude ash, fat, fibre, moisture, and starch were analysed for main ingredients (>10% inclusion rate) before formulation. Diets were produced by Research Diet Services (Wijk bij Duurstede, the Netherlands) and afterwards calculated nutrient values of diets were confirmed by near infrared spectroscopy (NIR). Diet and nutrient composition are presented in Table Bl.

Table 81: Diet composition (%) and nutrient concentrations (g / kg fresh diet, unless specified otherwise) for each phase.

Ingredients Starter 0 -10 d Wheat 31.8 Corn 30.0 Soybean meal 27.1 Soy protein concentrate 2.5 Sunflower meal 1.5 Soybean meal 2.5 Premix 0.5 L-Lysine 78% 0.3 DL-Meth. 99% 0.3 L-Threo. 98% 0.2 L-Valine 0.1 Limestone 1.3 Monocalciumphosphate 1.5 Sodium bicarbonate 0.4 NaCI 0.1 Xylanase 0.005 Maxiban 0.063 Nutrients -20 -Dry Matter 883 Crude Ash 60 Crude Protein 210 Crude Fat 42 Crude fibre 26 Starch 399 Calcium I 8.5 Phosphorus I 6.9 Sodium I 1.5 Digestible energy 2 2850 Lysine3 11.5 1Sum of digestible and indigestible minerals 2In kcal / kg 3Apparent fecal digestible lysine Broiler chickens Ross 308 one-day-old chicks (non-HatchCare) from a 47-week-old parent stock (reported 1st week offspring mortality: (0.74%) arrived from the hatchery at 0 d. Parent stock was vaccinated according to a standard vaccination schedule. On arrival, chicks were randomly divided over the pens and weighted on the pen level. The chicks were IC) vaccinated against Newcastle disease virus (0 and 14 d) and Gumboro disease (21 d). Chicks were confirmed to be free of Salmonella spp.

Broiler chicken measurements After placement, an ad random selection of 12 chicks was made and chickens were subjected to a chicken quality assessment consisting of measuring cloacal temperature, leg temperature, navel quality, and crop fill. Body weight (BVV) was measured on placement (0 d) and at 10 d, both BW and feed residual were measured. Animal performance data (BW; bodyweight, ADG; average daily gain, ADFI; average daily feed intake, and FCR; feed conversion ratio) were calculated for the starter phase (0 -10 d). No mortality corrections were done as no mortality was present in any of the pens. -21 -

Data analysis Data was recorded and stored afterwards in MS Excel on a OneDrive cloud-platform on pen level, which was also the experimental unit. After finishing the experiment, data was extracted from Excel into R (R Core Team 2019, version 3.6.1). Calculations, colour decoding, exploratory data analysis, data quality review, and subsequent statistical analyses were all performed in R. Linear mixed models were established for each parameter (ADG, ADFI, and FCR) separately, including treatments as fixed effects. Pen was added as random effect. If required, logarithmic transformation of the dependent variable was performed to meet model assumptions. If significant effects (P 0.05) were present, a post hoc Tukey test for pairwise comparison was performed on estimated marginal means to gain insight into differences between treatments. All data are presented as back-transformed estimated marginal means with standard error, unless stated otherwise.

3. Results and discussion Diet evaluation Diets were produced according to calculated nutrients.

Chick quality Chick quality was moderate to good at placement (data not shown). We measured cloaca temperature of broilers up to 5 d of age. Broilers reached target temperature after 1 d of age.

Broiler performance As presented in Table 32, feed stuff A enhanced ADFI (P = 0.04), which numerically also led to increased ADG in the feed stuff A group. However, effect sizes compared with control are rather small: 1.5 g, which means only 15 g greater feed uptake from placement up to 10 d of age. Feed conversion ratio was not affected by feed stuff A, as expected. Anice did not improve ADFI, ADG, or FCR, and might therefore not be suitable. However, higher inclusion rates may cause greater effect sizes, but this was not studied in this experiment. Body weight (Table B3) was not affected by treatments.

4. Conclusions

-22 -Feed stuff A enhanced feed intake during the brooding phase. Anice was not proven to enhance feed intake.

Table B2: Growth performance parameters of broiler chickens fed a control diet or supplemented with a taste enhancer. SEM = standard error of the mean. Different superscripts within a row represent between group differences, either significant (am,c; P 0.05) or tendencies (x,Y; P 5 0.10). Data are presented as estimated marginal means. n = 6 pens per treatment Phase Parameter Control Anice Feed Anice + SEM Treatment Stuff Feed effects' A Stuff A ADG 25.1 25 26.4 25.7 0.42 0.11 Starter ADFI 27.5" 27.6" 29.0Y 28.3"Y 0.39 0.04 0-10d FCR 1.09 1.10 1.10 1.10 0.006 0.68 'Model established P-values.

Table B3: Body weight of broiler chickens fed a control diet or supplemented with a taste enhancer. SEM = standard error of the mean. Data are presented as estimated 15 marginal means. n = 6 pens per treatment.

Age Control Anice Feed Anice + SEM Treatment effects' Stuff A Feed Stuff A 0 45 46 45 46 0.4 0.26 296 295 309 302 4.4 0.13 Model established P-values.

The invention thus provides methods of feeding animals and complementary feeds therefor.

Claims (18)

1. -23 -Claims 1. A supplementary feed, comprising a stoichiometric combination of GO 1 parts of one or more ribonucleotides; and (ii) 1 -10 parts glutamate.
2. 2. A supplementary feed according to claim 1, comprising a stoichiometric combination of (i) 1 parts of one or more ribonucleotides; and 00 1 -5 parts glutamate.
3. 3. A supplementary feed according to any previous claim, wherein the one or more ribonucleotides comprise disodium inosine-5'-monophosphate (IMP) or disodium guanosine-5'-monophosphate (GMP), or mixtures thereof.
4. 4. A supplementary feed according to claim 3, wherein the one or more ribonucleotides comprise a mixture of IMP and GMP at a ratio of approximately 1:1.
5. 5. A supplementary feed according to any preceding claim, wherein the supplementary 20 feed is a complementary feed.
6. 6. A supplementary feed according to any preceding claim, comprising: CO at least 1% by weight of one or more ribonucleotides; and (ii) at least 20% by weight of hydrolysed vegetable protein containing the 25 glutamate.
7. 7. A complementary feed according to claim 6, comprising: (i) at least 2% by weight of one or more ribonucleotides; and (ii) at least 40% by weight of hydrolysed vegetable protein containing the 30 glutamate.
8. 8. A supplementary feed according to claim 6 or 7, comprising: (i) up to 5% by weight of one or more ribonucleotides; and -24 - (ii) up to 90% by weight of hydrolysed vegetable protein containing the glutamate.
9. 9. A supplementary feed according to any of claims 1 to 8, comprising: (i) at least 50% by weight of a bulking agent.
10. 10. A supplementary feed according to any of claims 1 to 7, comprising: (i) at least 4% by weight of one or more ribonucleotides; (ii) at least 8% by weight of yeast or yeast extract containing the glutamate; and (iii) at least 60% by weight of a bulking agent.
11. 11 A supplementary feed according to claim 10, comprising: (i) at least 8% by weight of a mixture of IMP and GMP; (fi) at least 12% by weight of yeast or yeast extract containing the glutamate; (iii) at least 60% by weight of a bulking agent; and (iv) an anti-caking agent.
12. 12. A supplementary feed according to any previous claim, comprising: (i) 0.3% to 5% by weight of an anti-caking agent, and 20 wherein the feed is in the form of a dry powder.
13. 13. An animal feed, comprising up to 1% by weight of the supplementary feed of any previous claim.
14. 14. A method of modifying an animal feed, comprising: (i) providing an animal feed; (ii) combining (i) with up to 1% by weight of a supplementary feed according to any of claims ito 12.
15. 15. A method of feeding a non-human animal, comprising providing the animal with an animal feed according to claim 13, or providing the animal with an animal feed and supplementing the animal feed with up to 1% by weight of a supplementary feed according to any of claims 1 to 12.-25 - 16. A supplementary feed according to any of claims 1 to 12, an animal feed according to claim 13 or a method according to claim 14 or 15, wherein the animal is selected from: livestock, ruminants, pigs, fowl and equines.15. A supplementary feed according to any of claims 1 to 12, an animal feed according to claim 13 or a method according to claim 14 or 15, wherein the animal is a pig or a chicken.
16. 16. A supplementary feed according to any of claims 1 to 12, an animal feed according 10 to claim 13 or a method according to claim 14 or 15, wherein the animal is a dog or a cat.
17. 17. A method of feeding a pig, comprising providing the pig with an animal feed and supplementing the animal feed with from 0.05 to 0.1% by weight of a supplementary 15 feed according to any of claims 6 to 8 or claim 12 when dependent upon any of claims 6 to 8.
18. 18. A method of feeding a chicken, comprising providing the chicken with an animal feed and supplementing the animal feed with from 0.05 to 0.1% by weight of a 20 supplementary feed according to any of claims 6 to 8 or claim 12 when dependent upon any of claims 6 to 8.