Method for improving feed digestibility in bovine animals

Abstract

The present invention relates to a method for improving feed digestibility in bovine animals, in particular to the use of a combination of at least one bacterial amylase with a mixture of at least two essential oil compounds selected from the group consisting of thymol, eugenol, m-cresol, vanillin and guaiacol in the feed of bovine subfamily ruminants for improving weight gain, milk production and / or feed conversion ratio (FCR). Examples of bovine animals are beef cattle and dairy cattle.

Description

[0001] This application is a divisional application of Chinese patent application 201680009742.8 (PCT / EP2016 / 052957) filed on February 12, 2016. Invention Field

[0002] High-yielding cattle in modern agricultural systems live under conditions characterized by very high milk production (dairy cows) or growth rates (beef cattle), which require equally high energy demands. When intake increases beyond maintenance levels, feed utilization decreases significantly. In part to address this problem, increasingly biodegradable feeds are being incorporated into ruminant diets, such as starch-containing ingredients like grain-based concentrates and whole-grain silage. Starch-containing materials are frequently recovered in manure, meaning the utilization of such feed components can be further enhanced.

[0003] The energy content of bovine feed can be measured using an in vitro fermentation technique developed in 1979 by Professor Menke and his colleagues at the Department of Animal Nutrition, University of Hohenheim. The Hohenheim Feed Value Test (HFT) involves measuring the volume of gas produced during a 24-hour incubation of animal feed in rumen fluid. The amount of gas produced during incubation is directly related to feed digestibility and therefore to energy content. Since this method was first disclosed, numerous improvements and adjustments have been made, as described by Steingass and Menke in 1986.

[0004] Modifications to HFT allow for the observation of changes in gas productivity. Changes in certain aspects of the test, such as substrate type, substrate preparation, and / or incubation time, result in differences in the energy available to the substrate (feed) from the rumen fluid mixture. Adding certain substances to HFT fermentation can increase or decrease substrate digestibility. Therefore, there is a need for process consistency, such as the equipment and solutions used, the measurement procedures, and substrate preparation.

[0005] In animal feed, corn / maize or corn / maize silage is becoming increasingly important due to its high growth efficiency and energy-dense characteristics, especially in feed for ruminants.

[0006] Therefore, improvements are needed to enhance the digestibility of corn and / or starch in ruminant feeds to allow full utilization of energy potential and all available nutrients.

[0007] In addition, high-yielding cattle, characterized by high growth rates (beef cattle), are receiving diets with high levels of corn concentrate. This type of diet is widely used in farm cattle because it improves animal performance, carcass traits, and thus increases profit margins. This diet is often supplemented with monensin, a carboxyl ionotropic agent originally developed as an inhibitor of coccidiosis in poultry. Monensin is known to have beneficial growth-promoting properties when fed to cattle because it improves feed efficiency and prevents and controls parasitic infections in the herd.

[0008] A drawback of monensin is that it is a synthetic antibiotic. Therefore, there is a continued need to find sustainable alternatives for beef cattle that can reduce the use of antibiotics in animal husbandry and farming, and keep pace with the growing global demand for antibiotic-free meat products.

[0009] Related fields description WO 03 / 068256 A1 describes an amylase feed supplement for improving the nutrition of ruminants. The amylase used is a fungal amylase produced by Aspergillus oryzae. Tricarico et al. described the effects of Aspergillus oryzae extract containing α-amylase activity on rumen fermentation and milk yield in lactating Holstein cows in Animal Science 2005, 81:365-374.

[0010] Rojo et al. (Animal Feed Science and Technology, 123-124 (2005), 655-665) investigated the effects of exogenous amylases from Bacillus licheniformis and Aspergillus niger on rumen starch digestion and lamb performance. Invention Overview One object of the present invention is to provide an alternative, preferred improved concept that can alleviate the above-mentioned problems by improving feed utilization of farm animals, i.e. by improving feed conversion ratio and / or weight gain, and by increasing milk production in dairy cows.

[0011] Another object of the present invention is to provide an alternative feeding concept for farm animals that provides a sustainable method for producing animal meat products by replacing all or part of the antibiotics in the animal diet.

[0012] It has now been surprisingly discovered that the combined use of glycosylase and a mixture of at least two, preferably at least three, essential oil compounds in the feed for Bovidae farm animals has the advantage of significantly improving the digestibility of maize feed, said essential oil compounds being selected from the group consisting of thymol, eugenol, m-cresol, vanillin and guaiacol.

[0013] In particular, the inventors of this invention have discovered that supplementing with glycosylases (e.g., amylase) and mixtures of essential oil compounds selected from the group consisting of thymol, eugenol, m-cresol, vanillin, and guaiacol improves daily weight gain and feed efficiency in livestock herds.

[0014] A feedlot or pen is a type of animal husbandry operation used in intensive animal husbandry to raise (finish) livestock, especially beef cattle, as well as pigs, horses, sheep, turkeys, chickens, or ducks, before slaughter.

[0015] In the context of this invention, the Bovininae subfamily (also known as the genus *Bos* or bovine animals) refers to animals belonging to the kingdom Animalia, phylum Chordata, class Mammalia, order Artiodactyla, and family Bovidae. For the purposes of this invention, domestic cattle are the most preferred species. For the purposes of this invention, the term includes all breeds of domestic cattle, as well as cattle of all production types, particularly beef and dairy cattle.

[0016] It has also been found that a mixture of at least two, preferably at least three, essential oil compounds selected from the group consisting of thymol, eugenol, m-cresol, vanillin, and guaiacol in bovine animal feed can be used as a dietary alternative that can reduce or replace the amount of antibiotics currently used in farm animal feed while maintaining the important benefits of said antibiotics.

[0017] Therefore, in one embodiment, the present invention relates to a method for improving the digestibility of diets, such as those used for livestock in farms. More particularly, the present invention relates to a method for improving weight gain and / or feed conversion ratio (FCR) in beef cattle in a farm herd, the method comprising providing the animal with an effective amount of at least one glycoenzyme in combination with a mixture of at least two, preferably at least three, essential oil compounds selected from the group consisting of thymol, eugenol, m-cresol, vanillin, and guaiacol.

[0018] In another embodiment, the present invention relates to a method for sustainably producing meat products from bovine animals, the method comprising the steps of: formulating a feed composition intended for use in a farm by replacing all or part of the antibiotics in the composition with a mixture of at least two, preferably at least three, essential oil compounds selected from the group consisting of thymol, eugenol, m-cresol, vanillin, and guaiacol.

[0019] The term feed or feed composition refers to any compound, preparation, mixture or composition that is suitable for or intended for use by animals for ingestion.

[0020] The present invention also relates to a novel feed additive composition for farm animals, particularly beef cattle, comprising at least one glycoenzyme and a combination of at least two essential oil compounds as defined above as active ingredients. Invention Details The essential oil compounds according to the present invention are commercially available or can be prepared by a person skilled in the art using processes and methods well known in the prior art.

[0021] Essential oil compounds can be used in highly purified mixtures or as naturally available plant extracts or mixtures of extracts.

[0022] As used herein, the term "extract" includes compositions obtained by solvent extraction (also referred to as "extracted oils"), compositions obtained by steam distillation (also referred to as "essential oils"), or compositions obtained by other methods known to those skilled in the art. Suitable extraction solvents include alcohols, such as ethanol.

[0023] The term "natural" in this context is understood to mean a substance composed of naturally occurring compounds and obtained from natural products or through synthesis. Natural substances may preferably contain at least two of the compounds defined above as main components, as well as other essential oil compounds such as capsaicin, tannins, or carvacrol.

[0024] For beef cattle, the current consideration is to apply essential oil at a rate of 50 to 150 mg / kg body weight / day, preferably 70 to 120 mg / kg body weight / day (total dosage range of essential oil).

[0025] In another preferred embodiment of the invention, essential oils are added to feed as a single feed additive composition.

[0026] The essential oil-containing feed additive composition according to the present invention may optionally contain small amounts of other compounds, such as at least one compound present in plants, selected from the group consisting of: per kg of feed. Up to approximately 1 mg of propylidene, butylidene, phtalides, gingerol, and lavender oil; Up to approximately 2 mg of decanolactone, undecanolactone, doxydodecanolactone, ionone, irisone, eucalyptol, menthol, peppermint oil, and α-pinene; Up to approximately 3 mg of limonene, anethole, linalool, and methyl dihydrojasmonic acid; Up to approximately 4 mg of carvacrol, propionic acid, acetic acid or butyric acid, rosemary oil, clove oil, geraniol, terpineol, and citronellol; Up to approximately 5 mg of amyl salicylate and / or benzyl salicylate, cinnamaldehyde, plant polyphenols (tannins); And up to about 5 mg of turmeric powder or turmeric extract.

[0027] All essential oils and other compounds can be used in combination with emulsifying surfactants.

[0028] The emulsifier can be advantageously selected from those emulsifiers that are relatively hydrophilic, such as polyglycerol esters of fatty acids, such as esterified ricinoleic acid or propylene glycol esters of fatty acids, glycosyl esters or glycosyl glycerides, polyethylene glycol, lecithin, etc.

[0029] Examples of particularly preferred dosages of the essential oil compounds in the final feed additive composition according to the invention are independently within the following ranges: The concentration of thymol is between 80 and 120 g / kg, preferably 101 g / kg; Eugenol content is between 20 and 60 g / kg, preferably 30 g / kg; m-Cresol is between 80 and 110 g / kg, preferably 90 g / kg; The vanillin content is between 30 and 70 g / kg, with 50 g / kg being preferred; Guaiacin content is between 20 and 50 g / kg, preferably 35 g / kg; The salicylate / ester ratio is between 10 and 30, preferably 25 g / kg; The resorcinol content is between 5 and 20 g / kg, preferably 15 g / kg.

[0030] In a preferred embodiment of the feeding concept for beef cattle, the final feed comprises a mixture of thymol, m-cresol, and vanillin, wherein the three compounds are used in an amount sufficient to provide a daily dose of 50 mg to 150 mg of total essential oil per kg of the subject's body weight.

[0031] For the purposes of this invention, a preferred feed additive composition containing the claimed essential oil combination can be marketed under the trade name Crina. ® Ruminants (Crina) ® Crina (commercially available from DSM Nutritional Products AG, Kaiseraugst, Switzerland). ® Ruminants are blends of flavoring compounds used in animal nutrition and have a total essential oil content of 380 g / kg.

[0032] In the context of this article, glycosylases are enzymes that catalyze the breakdown of carbohydrates into monosaccharides.

[0033] Examples of glycoenzymes that can be used in the context herein are glucans, particularly β-glucanase and xyloglucanase, xylanase, amylase and pectinase, and mixtures thereof. In a preferred embodiment of the invention, the glycoenzyme is an amylase.

[0034] The glycoase used according to the present invention is stable in the presence of a protease. Protease stability can be determined by incubating 0.5 mg / ml of purified glycoase protein in a buffer at a desired pH (e.g., pH 3, 4, or 5) in the presence of a protease (e.g., pepsin, 70 mg / L) for a desired time (e.g., 30, 45, 60, 90, or 120 minutes), then raising the pH to the desired pH (e.g., pH 4, 5, 6, or 7) and measuring the residual activity. The residual glycoase activity relative to the control (untreated sample) is preferably at least 20%, preferably at least 30%, 40%, 50%, 60%, 70%, 80%, or at least 90%.

[0035] In one specific embodiment, the at least one glycoenzyme is an amylase or a mixture of enzymes, the enzyme composition comprising at least two enzymes selected from the group consisting of β-glucanase, xyloglucanase, xylanase, amylase and pectinase.

[0036] For the purposes of this invention, the preferred glycoenzyme is the glycoenzyme contained in the following commercial products: Ronozyme ® RumiStar ® Ronozyme ® VP, Ronozyme ® WX and Roxazyme ® (Available from DSM Nutrition Products AG in Kaiserlaust, Switzerland).

[0037] In the context of this paper, amylase is an enzyme that catalyzes the internal hydrolysis of starch and other linear and branched oligosaccharides and polysaccharides. In one specific embodiment, the amylase used according to the invention has α-amylase activity, that is, it catalyzes the internal hydrolysis of 1,4-α-glycosidic bonds in oligosaccharides and polysaccharides. α-Amylase acts randomly on, for example, starch, glycogen, and related polysaccharides and oligosaccharides, thereby releasing reducing groups in the α-configuration.

[0038] In a preferred embodiment, the amylase of the present invention is an α-amylase (systematic name: 1,4-α-D-glucan-glucan hydrolase), preferably a bacterial amylase. In another embodiment, the amylase of the present invention belongs to the EC 3.2.1 group of amylases, such as EC 3.2.1.1 (α-amylase), EC 3.2.1.2 (β-amylase), EC 3.2.1.3 (glucan 1,4-α-glucosidase, starch transglucosidase or glucosylamylase), EC 3.2.1.20 (α-glucosidase), EC 3.2.1.60 (glucan 1,4-α-maltotetrasaccharide hydrolase), EC 3.2.1.68 (isoamylase), EC 3.2.1.98 (glucan 1,4-α-maltohexanoase), or EC 3.2.1.133 (glucan 1,4-α-maltose hydrolase).

[0039] In a preferred embodiment, the amylase used according to the invention can be or is classified as belonging to group EC3.2.1.1. EC numbering references are made to Enzyme Nomenclature 1992, NC-IUBMB, Academic Press, San Diego, California, including supplements 1-5 disclosed in Eur. J. Biochem. 1994, 223, 1-5; Eur. J. Biochem. 1995, 232, 1-6; Eur. J. Biochem. 1996, 237, 1-5; Eur. J. Biochem. 1997, 250, 1-6; and Eur. J. Biochem. 1999, 264, 610-650. Nomenclature is periodically supplemented and updated; see, for example, the World Wide Web. http: / / www.chem.qmw.ac.uk / iubmb / enzyme / index.html .

[0040] Amylase activity can be determined by any suitable assay. Typically, the pH and temperature of the assay are adjusted for the enzyme under consideration. Examples of pH values ​​for measurement are pH 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. An example of a measurement temperature is 30°C. C, 35 C, 37 C, 40 C, 45 C, 50 C, 55 C, 60 C, 65 C, 70 C, 80 C, 90 C or 95 C. The preferred pH and temperature are within the physiological range, such as pH 3, 4, 5, 6, 7, or 8, and temperature 30°C. C, 35 C, 37 C or 40 C. The following amylase assay can be used: Substrate: Phadebas tablets (Pharmacia Diagnostics; cross-linked insoluble blue starch polymer, mixed with bovine serum albumin and buffer, and formulated into tablets). Assay temperature: 37°C C. pH determination: 4.3 (or 7.0 (if needed)). Reaction time: 20 minutes. After suspension in water, starch is hydrolyzed by α-amylase to yield soluble blue fragments. The absorbance of the blue solution measured at 620 nm is a function of α-amylase activity. One fungal α-amylase unit (1 FAU) is the amount of enzyme that breaks down 5.26 g of starch per hour under standard assay conditions. The preferred starch is soluble starch (Amylum solubile) Erg. B.6, batch 9947275, manufactured by Merck. More detailed description of the assay method APTSMYQI-3207 is available from Novozymes A / S, Krogshoejvej 36, DK-2880 Bagsvaerd, Denmark.

[0041] For the taxonomic classification and identification of bacteria, refer to Bergey's Manual of Systematic Bacteriology (1986), Vol. 2, ISBN 0-683-0783. Alternatively, well-known 16S rRNA sequence analysis can be used (see, for example, Johansen et al., Int. J. Syst. Bacteriol, 1999, 49, 1231-1240, especially the methods section in column 2 on page 1233); or consult a taxonomist, such as the DSMZ or other accredited depository.

[0042] As used herein, the term "bacterial" refers to an amylase derived from bacteria. The term "derived from" includes enzymes obtainable from wild-type bacterial strains and their variants. Such variants may have at least one substitution, insertion, and / or deletion of at least one amino acid residue. The term "variant" also includes shufflants, heterozygotes, chimeric enzymes, and consensus enzymes. Variants may have been generated by any method known in the art, such as site-directed mutagenesis, random mutagenesis, consensus derivatization (EP 897985), and gene shuffling (WO 95 / 22625, WO 96 / 00343), etc. For the purposes of this invention, an amylase variant is considered bacterial when at least one bacterial amylase has been used in its design, derivatization, or preparation. The term "bacterial" does not refer to a possible recombinant production host, but only to the origin of the amylase-encoding gene possessed by the recombinant production host.

[0043] The amylase used according to the present invention is preferably derived from Bacillus strains, such as Bacillus amyloliquefaciens, Bacillus circulans, Bacillus halmapalus, Bacillus licheniformis, Bacillus megaterium, Bacillus spp., Bacillus stearothermophilus, and Bacillus subtilis; preferably derived from strains of Bacillus amyloliquefaciens, Bacillus halmapalus, Bacillus licheniformis, Bacillus spp., Bacillus subtilis, and Bacillus stearothermophilus.

[0044] Non-limiting examples of wild-type amylases used according to the present invention are derived from the following wild-type amylases: *Bacillus licheniformis*, such as the Swissprot entry name AMY_BACLI, Master Accession Registry No. P06278; *Bacillus amyloliquefaciens*, such as the Swissprot entry name AMY_BACAM, Master Accession Registry No. P00692; *Bacillus megaterium*, such as the Swissprot entry name AMY_BACME, Master Accession Registry No. P20845; *Bacillus circulans*, such as the Swissprot entry name AMY_BACCI, Master Accession Registry No. P08137; and *Bacillus thermophilus*, such as the Swissprot entry name AMY_BACST, Master Accession Registry No. P06279. Another example is derived from *Bacillus subtilis*, such as the Swissprot entry name AMY_BACSU, Master Accession Registry No. P00691.

[0045] For the purposes of this invention, preferred amylases are those contained in the following commercial products: BAN, Stainzyme, Termamyl SC, Natalase, and Duramyl (all available from Novozymes), as well as Validase BAA and Validase HT (available from Valley Research). Other specific examples of amylases used according to the present invention are amylases contained in the following commercial products: Clarase, DexLo, GC 262 SP, G-Zyme G990, G-Zyme G995, G-Zyme G997, G-Zyme G998, HTAA, Optimax 7525, Purastar OxAm, Purastar ST, SpezymeAA, Spezyme Alpha, Spezyme BBA, Spezyme Delta AA, Spezyme DBA, Spezyme Ethyl, Spezyme Fred (GC521), Spezyme HPA, and Ultraphlow (all purchased from Genencor); Validase HT340L and Valley Thin 340L (both purchased from Valley Research); Avizyme 1500, Dextro 300 L, Kleistase, Maltazyme, Maxamyl, Thermozyme, Thermatex, and Starzyme. HT 120 L, Starzyme Super Conc and Ultraphlo.

[0046] In one specific embodiment, the amylase used according to the present invention is pelleting-stable and / or thermally stable. The melting temperature (Tm) of an enzyme is a measure of its thermal stability. The amylase of the present invention can have a melting temperature of at least 75°C. C, 76 C, 77 C, 78 C, 79 C, 80 C, 81 C, 82 C, 83 C, 84 C, 85 C, 86 C, 87 C、88 C, 89 C, 90 C, 91 C, 92 C, 93 C, 94 C or at least 95 The Tm of C was determined by differential scanning calorimetry (DSC). This DSC was performed in a 50 mM sodium chloride buffer solution (10 mM sodium phosphate, pH 7.0). The scan rate was constant, for example, 1.5. C / min. The scan interval can be from 20 to 100. C. Another buffer solution can be selected for the scan, for example, a buffer solution with pH 5.0, 5.5, 6.0, or pH 6.5. In another alternative implementation, a higher or lower scan rate can be used, for example, a lower one: 1.4. C / min, 1.3 C / min, 1.2 C / min, 1.1 C / min, 1.0 C / min or 0.9 C / min.

[0047] In another preferred embodiment, the amylase used according to the invention is at pH 7.0 and 37. The activity at C is relative to the optimal pH and 37. At least 35% of the activity at pH C. More preferably, at pH 7.0 and 37. The activity at C is at the optimal pH and 37 The activity at C is at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, or at least 75%.

[0048] In another preferred embodiment, the amylase of the present invention is used at pH 7.0 and 37. The activity exhibited in the presence of C and 5 mM bile salts is relative to that at the optimal pH and 37. C. At least 25% of the activity in the absence of bile salts. More preferably, at pH 7.0 and 37. The activity at the presence of C and 5 mM bile salts is optimal at pH 37. C. At least 30%, 35%, 40%, 45%, 50%, 55%, 60%, or at least 65% of the activity in the absence of bile salts.

[0049] The bacterial amylase used in this invention is the commercial product Ronozyme. ® RumiStar ® Active enzymes.

[0050] In one specific embodiment, the amylase is preferably defined in the form it is added to feed or when it is included in a feed additive. "Preferably defined" means that the enzyme preparation is at least 50% pure on a protein basis. In other specific embodiments, the enzyme preparation is at least 60%, 70%, 80%, 85%, 88%, 90%, 92%, 94%, or at least 95% pure. Purity can be determined by any method known in the art, such as by SDS-PAGE or by size exclusion chromatography (see Example 12 of WO01 / 58275).

[0051] Well-defined enzyme formulations are advantageous. For example, it is easier to correctly dosing enzymes, which are essentially unaffected or contaminating, into feed. The term "correctly dosing" specifically refers to the goal of obtaining consistent and constant results, and the ability to optimize the dosage based on the desired effect.

[0052] Enzyme preparations with this level of purity can be obtained using recombinant production methods, which are not readily available when produced using conventional fermentation methods and also exhibit much higher batch-to-batch variability.

[0053] The bacterial amylase used according to the present invention is contained in an effective amount in bovine diets or bovine feed additives. Currently, effective amounts below 200 mg enzyme protein / kg dietary dry matter are considered, preferably below 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 15, 10, 9, 8, or below 7 mg enzyme protein / kg dietary dry matter (ppm). On the other hand, effective amounts can be higher than 0.01 mg enzyme protein / kg dietary dry matter, preferably higher than 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.75, 1, 2, 3, or higher than 4 mg enzyme protein / kg dietary dry matter (ppm). Therefore, non-limiting examples of preferred dosage ranges are: 0.10 to 50 mg enzyme protein / kg, preferably 0.50 to 10, 1 to 9, 2 to 8, 3 to 8, or 4 to 7 mg enzyme protein / kg.

[0054] In the use according to the invention, the mixture of essential oil and amylase can be fed to the animal before, after, or simultaneously with its diet. The latter is preferred.

[0055] Feed conversion ratio (FCR) indicates how effectively feed is utilized. FCR can be determined based on animal growth trials, which include a first treatment in which a mixture of at least two compounds according to the invention is combined with amylase and added to the animal feed at an appropriate concentration per kg of feed, and a second treatment (control) in which no compounds are added to the animal feed.

[0056] As is generally known, the improved FCR is lower than the control FCR. In some specific embodiments, the FCR is improved (i.e. reduced) by at least 1.0% compared to the control, preferably at least 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, or at least 2.5%.

[0057] Improved weight gain refers to increased daily, weekly, bi-weekly, or monthly weight gain (in grams or kilograms for each relevant time period) relative to the control group that did not receive the addition of amylase and essential oils.

[0058] For feed compositions used in cattle, such as beef cattle, the cattle diet typically consists of an easily degradable fraction (referred to as concentrate) and a fiber-rich, less degradable fraction, which, according to the invention, comprises corn as a major component. Silage is a silage version of the fiber-rich fraction, obtained by ensiling material with high moisture content using a controlled anaerobic fermentation process (natural fermentation or additive treatment).

[0059] In addition to the amylase and essential oils as described above, the feed additive compositions of the present invention also contain at least one additional ingredient selected from vitamins and minerals. For example, the feed additives of the present invention may contain (i) at least one vitamin, (ii) at least one mineral, or (iii) at least one vitamin and at least one mineral.

[0060] The at least one vitamin can be either fat-soluble or water-soluble. Examples of fat-soluble vitamins are vitamin A, vitamin D3, vitamin E, and vitamin K such as vitamin K3. Examples of water-soluble vitamins are vitamin B12, biotin and choline, vitamin B1, vitamin B2, vitamin B6, niacin, folic acid, and pantothenic acid esters / salts, such as Ca-D-pantothenate.

[0061] At least one mineral can be a macromineral and / or a micromineral. Examples of microminerals are manganese, zinc, iron, copper, iodine, selenium, and cobalt. Examples of macrominerals are calcium, phosphorus, and sodium.

[0062] In practice, the incorporation of feed additive compositions as exemplified above into animal feed is done using concentrates or premixes. A premix represents a preferred homogeneous mixture of one or more trace ingredients with a diluent and / or a carrier. Premixes are used to promote the uniform dispersion of trace ingredients in a larger volume mixture. Premixes according to the invention can be added to feed ingredients or drinking water as a solid (e.g., as a water-soluble powder) or a liquid.

[0063] The premix may contain 0.5-10% by weight of the active ingredient according to the invention, and 10-95% by weight of other conventional additives, such as flavoring agents, vitamins, mineral salts, and any conventional absorbing support. The premix is ​​then added to the feed.

[0064] The present invention is further described through the following embodiments, which should not be construed as limiting the scope of the invention.

[0065] Example: Crina ® Ruminants and Ronozyme ® Rumistar ® Impact on Nelore cattle in feedlots Scheme and Design The study was conducted at the facility of the Department of Animal Science, ESALQ / USP (University of São Paulo), Pirasicaba-SP, Brazil.

[0066] The animals were divided into fifty pens, with six animals in each pen. Water was provided for free access, and feed was provided once daily. Cattle were fasted for 16 hours before body weight (BW) assessments were conducted at the beginning and end of the experiment. BW was assessed at 28 and 56 days to monitor partial average daily weight gain.

[0067] Treatment, number of animals and repetition Diet: High-concentration diet result The results are shown in Tables 1 and 2.

[0068] Table 1: Adaptation period – 28 days M+C = Monensin + Crina, C+R = Crina + Ronozyme Rumistar BW = Body weight, ADG = Average daily gain, DMI = Dry matter intake, FE = Feed efficiency In summary, compared to monensin, CRINA RUMINANTS increased average daily weight gain by 14.5% and feed efficiency by 7.34%. Compared to monensin, CRINA RUMINANTS + RONOZYME RUMISTAR (amylase) increased average daily weight gain by 23.3% and feed efficiency by 12.4%.

[0069] Table 2: After 59 days M+C = Monensin + Crina, C+R = Crina + Ronozyme Rumistar In summary, compared to monensin, CRINA RUMINANTS increased average daily weight gain by 14.7% and feed efficiency by 7.75%. Compared to monensin, CRINA RUMINANTS + RONOZYME RUMISTAR (amylase) increased average daily weight gain by 20.8% and feed efficiency by 10.9%.

Claims

1. A method for improving weight gain and / or feed conversion ratio in bovine animals, characterized in that... The animal is given an α-amylase of at least one enzyme of group EC 3.2.1.1 and a mixture of at least two, preferably at least three, essential oil compounds selected from the group consisting of thymol, eugenol, m-cresol, vanillin and guaiacol, wherein the amylase is a bacterial amylase and wherein the animal is a beef cattle.

2. The method according to claim 1, wherein the essential oil is applied at a dose of 50 to 150 mg / kg body weight / day (total dose range of the essential oil).

3. The method according to claim 1, wherein the essential oil is applied at an amount of 70 to 120 mg / kg body weight / day (total dose range of the essential oil).

4. The use of at least one α-amylase of the EC 3.2.1.1 group and a mixture of at least two, preferably at least three, essential oil compounds in the feed of bovine animals for improving weight gain and / or feed conversion ratio, wherein the essential oil compounds are selected from the group consisting of thymol, eugenol, m-cresol, vanillin and guaiacol, wherein the amylase is a bacterial amylase, and wherein the animal is a beef cattle.

5. The use according to claim 4, wherein the essential oil is applied at a dose of 50 to 150 mg / kg body weight / day (total dose range of the essential oil).

6. The use according to claim 4, wherein the essential oil is applied at a dose of 70 to 120 mg / kg body weight / day (total dose range of the essential oil).

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