WO2020189916A1 - Low-protein feed composition for promoting feed conversion efficiency of ruminants - Google Patents

Abstract

The present invention pertains to a low-protein feed composition for promoting the feed conversion efficiency of ruminants, the low-protein feed composition containing: a carbohydrate source; a protein source containing soybean meal; a fat source; and β-mannanase as an enzyme, wherein the content of the soybean meal is in the range of 4.0-7.0 parts by weight with respect to 0.1 parts by weight of the β-mannanase. In the low-protein feed composition, which has a lower protein content than conventional feed compositions, the mixing ratio of soybean meal and β-mannanase is optimized to reduce the amount of soybean meal used. Thus, the low-protein feed composition has the effect of increasing feed conversion efficiency (FCE) while having good economic feasibility.

Description

Low protein feed composition for enhancing feed conversion efficiency of ruminants

The present invention relates to a feed composition, and in particular, to a low protein feed composition for enhancing feed conversion efficiency of ruminants.

Throughout the history of humanity, livestock has formed an inseparable relationship by providing livestock products such as meat, milk, and eggs. In particular, the meaning is deeper in the aspect that livestock feeds in low-quality plant feed ingredients and supplies us with protein sources. Therefore, attempts to increase the productivity of livestock have been steadily made, and rapid progress is being made through advanced technology.

According to the Ministry of Agriculture, Food and Rural Affairs, more than 50% of livestock production costs are attributed to feed costs. Therefore, it goes without saying that feed cost reduction is a big task in livestock management. In the case of feed containing a large amount of mannan (palm meal, copra meal, soybean husk, etc.), along with agri-food by-products, feed costs were reduced and the ruminants contributed greatly as a protein ingredient. Unlike unit animals, in ruminants, fibrinolytic bacteria present in the rumen can decompose beta-mannans. However, due to the anti-nutritional factors of feeds with high met content, that is, low digestibility, low energy content, and palatability problems, the amount of feed intake is lowered, and the amount of use thereof is limited.

Meanwhile, the enzyme field can be said to be one of the most spotlighted fields in the last 20 years, and the main purpose of the use of enzymes is to improve digestibility of low-quality feed using enzymes and to remove anti-nutritional factors (ANFs). The grafting of these enzymes in livestock is helping to remove anti-nutritional factors in feed and improve digestibility, as can be seen from phytase, xylanase, and β-glucanase, which are currently most widely used.

However, in the case of ruminants, the addition of enzymes in feed is not receiving much attention because the microorganisms in the rumen produce most of the enzymes. Most of the enzymes currently used in Mimihanama ruminants are designed to help break down fibrin. However, it is not an exaggeration to say that the addition of external enzymes is actually faced with many limitations due to the nature of the rumen due to the interactions between microorganisms, symbiotic reactions, and chain reactions caused by secretions.

In this regard, the inventors of the present invention in the Republic of Korea Patent Publication No. 10-2016-0061539 (name of the invention: beta-met kinase-containing ruminant feed enzyme preparation), 12 weeks of age or more ruminant developed ruminant animal feed for sex or rumination It is administered directly to the animal adult, and a composition containing beta-mannanase (β-mannanase), a feed enzyme preparation for ruminants was presented. Through this, the addition of beta metase improves the growth rate, feed efficiency, and nitrogen utilization rate in ruminants, thereby improving feed efficiency and raw material availability when growing ruminants, and inexpensive raw materials such as soybean meal, palm meal, palm meal, and alcoholic beverages can be used. It has been suggested that economic feasibility can be improved.

However, in the Korean Patent Application Publication No. 10-2016-0061539 described above, it is not described to what extent an ingredient that has a great influence on economic efficiency such as soybean meal can be used, and if it does not affect the specific flow rate and milk ingredient Therefore, the mixing ratio of protein sources such as beta mannase and soybean meal, which can increase feed conversion efficiency, was still unknown.

The present invention is to solve the above problems, in a low protein feed composition having a lower protein content than the conventional, it is an object to improve the feed conversion efficiency (FCE: Feed Conversion Efficinecy) while excellent economical efficiency.

In addition, the present invention reduces the content of soybean meal, which has the greatest effect on economics among the protein sources in feed, so that even if the content of Crude Protein (CP) is low, feed conversion efficiency (FCE) is increased, and the specific flow rate and milk component are affected. It is intended to provide a feed composition capable of lowering the urea mass (MUN) and somatic cell count without having to do so.

The present invention for achieving the above object, corn silage (Corn silage), steam-flaked flakes (Steam-flaked corn), and at least one carbohydrate source selected from the group consisting of flat barley rice (Rolled barley); A protein source including soybean meal, and further comprising one or more selected from the group consisting of Alfalfa hay, Distillers grain, and Soybean hulls; A local source of cotton seeds; And beta-mannanase (β-mannanase) as an enzyme, wherein the soybean meal is contained within the range of 4.0 to 7.0 parts by weight based on 0.1 parts by weight of the beta-metase, feed conversion efficiency of ruminants (FCE: Feed Conversion Efficinecy) is a low protein feed composition for enhancement.

And, in the feed composition according to the present invention, the crude protein (CP: Crude Protein) may be contained within the range of 14.5 to 15.0% by weight.

In addition, as an example, the soybean meal may be included in the range of 5.0 to 5.5 parts by weight based on 0.1 parts by weight of the beta met nanase.

In addition, as an example, the carbohydrate source contains corn silage 25.0 to 26.0 parts by weight, steam-flaked corn 11.0 to 12.0 parts by weight, and rolled barley 7.0 to 8.0 parts by weight. It is possible that it has been made.

In addition, as an example, the protein source is 4.0 to 7.0 parts by weight of Soybean meal, 26.0 to 27.0 parts by weight of Alfalfa hay, 5.0 to 6.0 parts by weight of Distillers grain, and Soybean meal. hulls) 11.0 to 12.0 parts by weight may be included.

In addition, as an example, the fat source may be made by including 6.0 to 7.0 parts by weight of cotton seed.

In addition, as an example, the present invention may further include at least one selected from the group consisting of mineral mix, calcium carbonate, and salt (NaCl).

In addition, as an example, it is possible to further include 1.0 to 2.0 parts by weight of the mineral mix, 0.1 to 0.2 parts by weight of calcium carbonate, and 0.1 to 0.2 parts by weight of salt (NaCl). .

In addition, as an example, the present invention may be to increase feed conversion efficiency and lower urea mass (MUN) and somatic cell count compared to a feed composition that does not contain beta-mannanase as an enzyme. .

On the other hand, the low protein feed composition for improving the feed conversion efficiency (FCE) of ruminants according to the present invention, corn silage 25.0 to 26.0 parts by weight, steam-flaked corn 11.0 to 12.0 A carbohydrate source comprising 7.0 to 8.0 parts by weight, and rolled barley; Soybean meal 4.0 to 7.0 parts by weight, Alfalfa hay 26.0 to 27.0 parts by weight, Distillers grain 5.0 to 6.0 parts by weight, and Soybean hulls 11.0 to 12.0 parts by weight. Protein source; Fat source consisting of 6.0 to 7.0 parts by weight of cotton seed; And 0.1 parts by weight of beta-mannanase as an enzyme.

In addition, 1.0 to 2.0 parts by weight of a mineral mix, 0.1 to 0.2 parts by weight of calcium carbonate, and 0.1 to 0.2 parts by weight of salt (NaCl) may be further included.

In addition, the low-protein feed composition for improving feed conversion efficiency of ruminants according to the present invention includes 25.65 parts by weight of corn silage, 11.3 parts by weight of steam-flaked corn, and 7.21 parts by weight of rolled barley. A carbohydrate source comprising parts by weight; A protein source comprising 5.21 parts by weight of Soybean meal, 26.2 parts by weight of Alfalfa hay, 5.01 parts by weight of Distillers grain, and 11.43 parts by weight of Soybean hulls; Fat source consisting of 6.55 parts by weight of cotton seed; 0.1 parts by weight of beta-mannanase as an enzyme; 1.09 parts by weight of mineral mix; 0.16 parts by weight of calcium carbonate; And it is more preferable to include 0.19 parts by weight of salt (NaCl).

Details of other embodiments are included in the detailed description and drawings.

The present invention is a low protein feed composition with a lower protein content than the conventional, by optimizing the mixing ratio of soybean meal and beta-mannanase (β-mannanase), while reducing the amount of soybean meal, excellent economic efficiency, feed conversion efficiency (FCE: Feed Conversion Efficinecy) can be increased.

In addition, the present invention reduces the content of soybean meal, which has the greatest effect on economics among the protein sources in feed, so that even if the content of Crude Protein (CP) is low, feed conversion efficiency (FCE) is increased, and the specific flow rate and milk component are affected. It is possible to provide a feed composition capable of lowering urea mass (MUN) and somatic cell count without having to do so.

In the present invention, various transformations may be applied and various embodiments may be provided, and specific embodiments will be illustrated in the drawings and described in detail in the detailed description. However, this is not intended to limit the present invention to a specific embodiment, it is to be understood to include all conversions, equivalents, and substitutes included in the spirit and scope of the present invention. In describing the present invention, when it is determined that a detailed description of a related known technology may obscure the subject matter of the present invention, a detailed description thereof will be omitted.

The terms used in the present application are only used to describe specific embodiments, and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In the present application, terms such as "comprise" or "have" are intended to designate the presence of features, numbers, steps, actions, components, parts, or combinations thereof described in the specification, but one or more other features. It is to be understood that the presence or addition of elements or numbers, steps, actions, components, parts, or combinations thereof, does not preclude in advance.

Terms such as first and second may be used to describe various components, but the components should not be limited by the terms. These terms are used only for the purpose of distinguishing one component from another component.

The present invention relates to a feed composition, particularly to a feed composition that can be added to a ruminant feed or directly administered to a ruminant, and more particularly, to a low protein feed composition for enhancing feed conversion efficiency of ruminants. .

In the present invention, "ruminant" is meant to include cattle, sheep, goats, camels, water buffalo, deer, reindeer, caribou (caribou) and elk having a stomach having a complex and many rooms.

The low protein feed composition for enhancing feed conversion efficiency (FCE) of ruminants according to the present invention is composed of corn silage, steam-flaked corn, and rolled barley. One or more carbohydrate sources selected from the group; A protein source including soybean meal, and further comprising one or more selected from the group consisting of Alfalfa hay, Distillers grain, and Soybean hulls; A local source of cotton seeds; And beta-mannanase (β-mannanase) as an enzyme, and the soybean meal is contained in the range of 4.0 to 7.0 parts by weight based on 0.1 parts by weight of the beta-met nanase.

In the present invention, beta-mannanase (β-mannanase) lowers the viscosity in the digestive tract to improve the digestibility of various nutrients, causes energy loss through stimulation of the immune system shown by the anti-nutritional factor, mannan, and interferes with the secretion of insulin and igf-1. The side can be removed. The beta met kinase can be obtained from a microorganism that produces a met kinase or a transformant transduced with a met kinase gene. In one example, a beta metase capable of exhibiting an effect without being decomposed even in the stomach of a ruminant is preferred, and CTC Bio's CTCZYME ^® product may be used, but is not limited thereto. The beta-met nanase is endo-β-mannanase, and can be obtained from a microorganism or a transformant transduced with a met- kinase gene. For example, a) a metase produced from the Bacillus sp WL-1 (Bacillus sp WL-1) strain disclosed in Korean Patent Registration No. 10-0390565 and deposited with accession number KCTC 0800BP; b) a metase produced from Bacillus sp WL-7 (Bacillus sp WL-7) strain disclosed in Korean Patent Registration No. 10-0477456 and deposited with accession number KCTC 10279BP; c) a metase produced by expression of E. coli transformed by introducing a metase gene of Bacillus licheniformis WL-12 strain disclosed in Korean Patent Registration No. 10-0560375; And d) a metase produced from Aspergillus duck material disclosed in Korean Patent Registration No. 10-1073987 and deposited with accession number KCTC 11105BP; e) a metase produced from the Cellulosimicrobium sp HY-13 strain disclosed in Korean Patent Registration No. 10-1403489 and deposited with the accession number KCTC 11302BP; And f) it may be selected from a mixture of met kinase selected from a) to e). The contents of the Korean patents in which each of the above beta met nanases are disclosed are incorporated by reference in their entirety into the contents of the present invention.

In general, the addition of exogenous enzymes into the feed allows animals to use the nutrients in the feed to the fullest, thus reducing the environmental burden of pigs, poultry and even cattle by broadening the range.

The present inventors compared and evaluated the feed conversion efficiency (Feed Conversion Efficiency) and the relative flow rate for cattle ingesting a standard feed and a feed containing beta met kinase added to the feed with low crude protein content, To investigate the effect of.

First, twelve intermediate lactating fertility Holstein cows producing 40.5±3.6 kg milk per day were divided into three experimental groups and a 3x3 crossover design was conducted. The experimental groups were as follows: a complete mixed diet containing high crude protein (16.1% crude protein, high crude protein, HCP), a complete mixed diet containing low crude protein (14.6% crude protein, low crude protein, LCP) and 0.1% low crude protein. Dry matter (DM, dry matter) Mixed feed with beta metase enzyme added (feed with low crude protein enzyme added, LCPE1).

The addition of beta met nanase had no effect on dry matter intake (DMI), milk yield, and milk component yield or composition. Milk urea nitrogen (MUN) was significantly lower in cows fed LCP (low crude protein feed) or LCPE1 (low crude protein feed supplemented with beta-metinase). In the case of somatic cell count, it was found that the level was lower in cows fed diet (LCPE1) containing beta-met kinase than the other two experimental groups (LCP, HCP). Dry matter (DM), organic matter (OM), crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), carbohydrate (starch), There was no effect on the apparent total tract digestibility (ATTD) of the total energy of ash. Cows fed LCPE1 had 190g higher feed conversion efficiency (FCE) per kg of dry matter intake (13.4% synergistic effect, P = 0.003) than cows fed HCP. Compared with cows fed with LCP feed, it was found that the feed conversion efficiency (FCE) was significantly higher ( P = 0.014) in cows fed LCPE1 (160g per kg of dry matter, 11.0% increase). Cows fed LCPE1 use crude protein more efficiently in synthesizing milk protein than cows fed HCP feed (milk protein: crude protein intake = 0.34 vs. 0.30). Due to the difference in the crude protein content in the feed, urea intake was significantly lower in cows with low crude protein intake, and therefore the amount of urea escaped in feces and urine was also small, which in turn minimized the impact on the environment. In cows with 0.1% dry matter intake, beta metase improved feed conversion efficiency (FCE) and had no effect on specific flow and dairy components (in cows), but instead had no effect on the number of somatic cells and the amount of urea released to the environment. Lowered.

As described above, the present inventors conducted an experiment to see how beta met nanase affects cows when providing feed to which thrombolytic enzyme (beta met nanase) was added to lactating cows. As a result, the protein content was high ( The feed conversion efficiency was higher in cows that ate feed (LCPE1) with beta metase added to feed with lower protein content (14.6% of dry matter) than cattle fed with dry matter 16.1%) feed (HCP). It was confirmed that the addition did not affect the specific flow rate and the components of milk, but only lowered the somatic cell count.

Next, the inventors of the present invention are to determine to what extent components that have a large impact on economic efficiency such as soybean meal in the feed composition can be used, and to maximize feed conversion efficiency (FCE) without affecting the specific flow rate and milk components. To find out the mixing ratio of beta mannase, feed (LCPE2) with lower soybean meal content as a protein source and higher content of Distillers grain (LCPE2) as a protein source, and lower soybean meal content as a protein source and as a source of carbohydrate. Additional experiments were also conducted for feed (LCPE3) with increased steam-flaked corn content.

As a result, the feed conversion rate slightly increased in the case of feed with increased Distillers grain content (LCPE2) compared to feed with beta metase added to feed with low protein content (14.6% of dry matter) (LCPE1). It stopped, and there was a drawback of reducing the specific flow rate. In addition, in the case of feed (LCPE3) with increased steam-flaked corn content, the crude protein (CP) content was significantly lowered, the specific flow rate was also reduced, and there was a disadvantage that a large change occurred in the milk component.

Accordingly, the low-protein feed composition for enhancing feed conversion efficiency (FCE) of ruminants according to the present invention includes corn silage, steam-flaked corn, and rolled barley. ) At least one carbohydrate source selected from the group consisting of; A protein source including soybean meal, and further comprising one or more selected from the group consisting of Alfalfa hay, Distillers grain, and Soybean hulls; A local source of cotton seeds; And beta-mannanase (β-mannanase) as an enzyme, and the soybean meal is contained in the range of 4.0 to 7.0 parts by weight based on 0.1 parts by weight of the beta-met nanase.

As an example, it is more preferable that the soybean meal is contained within the range of 5.0 to 5.5 parts by weight based on 0.1 parts by weight of the beta met nanase. If soybean meal is included below the above range, there is a disadvantage in that the crude protein (CP) content is significantly lowered, the specific flow rate is also reduced, and a large change occurs in the milk component. In addition, if the soybean meal is included in excess of the above range, there is a disadvantage in that the economy is poor.

And, in the feed composition according to the present invention, the crude protein (CP: Crude Protein) may be contained within the range of 14.5 to 15.0% by weight. That is, the feed composition according to the present invention is characterized by having the maximum effect of an enzyme by beta mannase in a low protein feed composition having a relatively low crude protein content by lowering the content of the protein source to the maximum.

The present invention is a low protein feed composition with a lower protein content than the conventional, by optimizing the mixing ratio of soybean meal and beta-mannanase (β-mannanase), while reducing the amount of soybean meal, excellent economic efficiency, feed conversion efficiency (FCE: Feed Conversion Efficinecy) can be increased.

In addition, the present invention reduces the content of soybean meal, which has the greatest effect on economics among the protein sources in feed, so that even if the content of Crude Protein (CP) is low, feed conversion efficiency (FCE) is increased, and the specific flow rate and milk component are affected. It is possible to provide a feed composition capable of lowering urea mass (MUN) and somatic cell count without having to do so. Thus, the present invention may be to increase feed conversion efficiency and lower urea mass (MUN) and somatic cell count, compared to a feed composition that does not contain beta-mannanase as an enzyme. This invention has the effect of saving $1.03 per cow when using LCPE1 feed rather than HCP feed.

According to the experimental example of the present invention, as an example, the carbohydrate source is corn silage 25.0 to 26.0 parts by weight, steam-flaked corn 11.0 to 12.0 parts by weight, and rolled barley 7.0 It is possible to include to 8.0 parts by weight. When the content of a carbohydrate source such as steam-flaked corn is increased, the crude protein (CP) content is significantly lowered, the specific flow rate is also reduced, and there is a disadvantage that a large change occurs in the milk component.

In addition, as an example, the protein source is 4.0 to 7.0 parts by weight of Soybean meal, 26.0 to 27.0 parts by weight of Alfalfa hay, 5.0 to 6.0 parts by weight of Distillers grain, and Soybean meal. hulls) 11.0 to 12.0 parts by weight may be included. If the content of alcoholic beverages rather than soybean meal is increased, there is a disadvantage that the increase in feed conversion rate is not large and the specific flow rate decreases.

In addition, as an example, the fat source may be made by including 6.0 to 7.0 parts by weight of cotton seed.

In addition, the feed composition according to the present invention may further include one or more selected from the group consisting of mineral mix, calcium carbonate, and salt (NaCl).

In addition, as an example, it is possible to further include 1.0 to 2.0 parts by weight of the mineral mix, 0.1 to 0.2 parts by weight of calcium carbonate, and 0.1 to 0.2 parts by weight of salt (NaCl). .

The mineral mix refers to a mixture prepared in advance to facilitate mixing of minerals required for a ruminant with raw materials when mixing feed. Minerals form the skeleton in the body of ruminants and can be used as important nutrients in metabolic processes. The amount of minerals required may vary depending on the degree of distribution in the body of livestock. Usually, the required amount of Ca, P, Na, Cl, K, and Mg is large, and the required amount of Fe, Zn, I, Se, Mn, Cu, etc. Is known to be a trace amount, but in ruminant feed, K, Mg, Mn, S, etc. are sufficiently contained in the feed and are not insufficient, but other minerals must be added to the feed. In the present invention, a commercially available product may be used for the mineral mix, and an appropriate mixing ratio of each mineral is known in the art. In one embodiment of the present invention, the mineral mix is preferably included in the range of 1.0 to 2.0 parts by weight.

The salt is used as a source of sodium (Na) and chlorine (Cl) in ruminant feed compositions. Sodium and chlorine are the major extracellular cations and anions in the body, respectively. Chlorine is the main anion in digestive juices. Salt can meet the sodium and potassium requirements of ruminants in feeds based on corn and soybean meal. Deficiency of sodium or chlorine can slow the growth of ruminants and reduce feed efficiency. In the present invention, salt may preferably be included in an amount of 0.1 to 0.2 parts by weight.

In addition, the feed composition according to the present invention is a vitamin premix; One or more sources of minerals selected from calcium phosphate, limestone, and dephosphorylated ores; molasses; And it may further include one or more selected from the group consisting of amino acids.

On the other hand, the low protein feed composition for improving the feed conversion efficiency (FCE) of ruminants according to the present invention, corn silage 25.0 to 26.0 parts by weight, steam-flaked corn 11.0 to 12.0 A carbohydrate source comprising 7.0 to 8.0 parts by weight, and rolled barley; Soybean meal 4.0 to 7.0 parts by weight, Alfalfa hay 26.0 to 27.0 parts by weight, Distillers grain 5.0 to 6.0 parts by weight, and Soybean hulls 11.0 to 12.0 parts by weight. Protein source; Fat source consisting of 6.0 to 7.0 parts by weight of cotton seed; And 0.1 parts by weight of beta-mannanase as an enzyme.

In addition, 1.0 to 2.0 parts by weight of a mineral mix, 0.1 to 0.2 parts by weight of calcium carbonate, and 0.1 to 0.2 parts by weight of salt (NaCl) may be further included.

In addition, the low-protein feed composition for improving feed conversion efficiency of ruminants according to the present invention includes 25.65 parts by weight of corn silage, 11.3 parts by weight of steam-flaked corn, and 7.21 parts by weight of rolled barley. A carbohydrate source comprising parts by weight; A protein source comprising 5.21 parts by weight of Soybean meal, 26.2 parts by weight of Alfalfa hay, 5.01 parts by weight of Distillers grain, and 11.43 parts by weight of Soybean hulls; Fat source consisting of 6.55 parts by weight of cotton seed; 0.1 parts by weight of beta-mannanase as an enzyme; 1.09 parts by weight of mineral mix; 0.16 parts by weight of calcium carbonate; And it is more preferable to include 0.19 parts by weight of salt (NaCl).

According to the examples and experimental examples of the present invention below, as a result of adding beta-metinase to corn silage (livestock feed stored in silos using corn stems and leaves as raw materials) and alpha waves, fertility Holstein cows in the intermediate lactation stage are produced. The production of one milk flow increased while the number of somatic cells decreased. The feed conversion efficiency (FCE) value in the feed was more improved when the beta metase was added rather than lowering the forage ratio in the feed to the maximum. When beta met nanase was added to 0.1% dry matter, the amount of nitrogen in the milk urea was significantly decreased due to the decrease in nitrogen intake, although it did not have a significant effect on the flow rate and milk composition. There was no change in the apparent total tract digestibility of DM, OM, CP, ADF, NDF, and starch. The amount of nitrogen found in feces and urea also decreased as the amount of nitrogen intake was reduced in cows fed a low crude protein diet. When urea is released into the environment, it is highly likely to be converted to ammonia or nitrogen dioxide, so reducing nitrogen in urea is beneficial not only to the environment but also to (bovine) health. If the ratio of crude protein in the diet is further reduced, the possibility that the number one is protected by amino acids is also conceivable. Therefore, we need to further study how much less crude protein given to cattle does not affect cattle flow or cattle health. What we found through our experimental research is that even if the ratio of crude protein in the feed is lowered, the feed conversion efficiency (FCE) can be improved, and the number of somatic cells can be decreased independently of the flow rate. . If the ratio of forage in the feed is lowered and beta metase is added instead, a profit of $1.03 per cow could be made.

The present invention may be better understood by the following examples, and the following examples are for illustrative purposes of the present invention, and are not intended to limit the scope of protection defined by the appended claims.

Example 1: Experimental animals and experimental group

All animal-related experiments were approved by the Laboratory Animal Steering Committee located at UC Davis. This study was conducted at the animal science education and research development facility located within UC Davis University. Twenty mid-breasting Holstein cows producing 40.5 ± 3.6 kg milk per day have an average weight of 643 ± 48 kg BW, an average specific flow of 40.5 ± 3.6 kg per day, and 128 ± 9.64 Dried Fish (DMI). Were kept in open cages (American Calan, Northwood, NH) equipped with Calm fences at the beginning of the experiment. The experimental groups were as follows: a complete mixed diet containing high crude protein (16.1% crude protein, high crude protein, HCP), a complete mixed diet containing low crude protein (14.6% crude protein, low crude protein, LCP), 0.1% low crude protein. Dry matter (DM, dry matter) mixed feed with beta metase enzyme added (feed with low crude protein enzyme added, LCPE1), feed with lower soybean meal and higher Distillers grain as a protein source (LCPE2), and feed with lower soybean meal content as a protein source and higher steam-flaked corn content as a carbohydrate source (LCPE3).

Beta metase enzyme CTCZYME (Korea Patent Registration 10-0477456, CTC Bio) contains pure isolated beta metase. The beta-met kinase was generated by repeating replication after inserting Bacillus subtills (WL-7) bacteria encoding the met kinase gene into Escherichia coli , and then repeating the replication (met kinase produced by E. coli expression). The metase gene encodes the 362 amino acids constituting the polypeptide, and this construct is identical to the metase belonging to the GH family 26 taxon. Enzymatic reaction was carried out at 800,000 U/kg in pH 4.0 and 30 ^o C environment.

Constituents and chemical elements of the feed used in the experiment are listed in Tables 1 and 2 below.

At first, the enzyme was hand-mixed in the soybean silage and given to the animals. Then, the soybean silage, which had already been mixed first, was mixed with the rest of the feed. The experimental cycle of each experiment was 18 days, 14 days was a period of adaptation in a Pristol barn, one day was a period of adaptation to metabolic changes, and urine and feces from each cow were collected for the remaining 3 days. The metabolic stall consisted of food, drinking cups, and rubber soles. Cows in the Pristol barn and metabolic stall were given 110% of the amount of feed consumed the previous day, twice a day at 8 am and 8 pm. Relative flow, dairy composition, and DMI were measured for each cow living in a Pristol barn and metabolic stall. Water supplies were provided to all animals from time to time.

Ingredients (% of DM)	HCP	LCP	LCPE1	LCPE2	LCPE3
Corn silage 1	24.62	25.65	25.65	25.65	25.65
Alfalfa hay 2	25.3	26.2	26.2	26.2	26.2
Steam-flaked corn	10.5	11.3	11.3	11.3	12.8
Distillers grain	6.47	5.01	5.01	6.51	5.01
Soybean meal	7.94	5.21	5.21	3.71	3.71
Rolled barley	8.46	7.21	7.21	7.21	7.21
Mineral mix	1.09	1.09	1.09	1.09	1.09
Soybean hulls	10.04	11.43	11.43	11.43	11.43
Cotton seed	5.23	6.55	6.55	6.55	6.55
Calcium carbonate	0.16	0.16	0.16	0.16	0.16
Salt (NaCl)	0.19	0.19	0.19	0.19	0.19
β-mannanase enzyme	0	0	0.1	0.1	0.1
Sum	100	100	100.1	100.1	100.1

¹ Contained 46.2% DM and 7.9% CP, 38.9% NDF, and 30.5% starch on DM basis. ² Contained 88.1% DM and 24.2% CP, 33.5% NDF, and 2.8% starch on DM basis.

Chemical composition (% of DM)	HCP	LCP	LCPE1	LCPE2	LCPE3
Building (DM: dry matter)	64.6	64.9	64.9	64.9	64.9
Crude Protein (CP)	16.1	14.6	14.6	14.4	12.9
Acid Detergent Fiber Fraction (ADF)	27.5	25.7	25.7	25.6	24.8
Neutral Detergent Fiber Fraction (NDF)	38.9	36.5	36.5	36.6	37.1
Lignin	4.6	4.1	4.1	4.2	4.3
Starch	21.8	22.2	22.2	22.3	24.1
Ether Extract	4.1	4.3	4.3	4.3	4.2
Sum	113	107.4	107.4	107.4	107.4

As shown in Table 2, in the case of a low crude protein feed (LCP), a mixed feed in which a beta metase enzyme is added to a low crude protein (LCPE1), and a feed with a high Distillers grain content (LCPE2), crude protein The content of (CP) was similar, but the content of crude protein (CP) was significantly lowered in the feed (LCPE3) with increased content of steam-flaked corn, a source of carbohydrates.

Example 2: Sample collection and analysis

The amount of feed consumed by cattle or not consumed was weighed daily.

The period was set and the remaining feed was sampled daily. Cattle feed was analyzed on the eve of Day 13 and at the end of Day 18. Feed components and several days of feed remaining from cows were collected and stored at -20 ⁰ C until chemical composition review. The cows are milked twice daily at 6 am and 6 pm. Milk samples from one cow each day were stored at 4 ^° C until milk fat, milk protein, lactose, milk nitrogen concentration (MUN), and milk somatic (SCC) analysis.

From Day 16 to Day 18, the amount of feces and urine per bovine (kg/d) in a metabolic stall was measured. I guess, digestion and metabolic reactions by enzymes might reach homeostasis in the body after a two-week adaptation period. Before the cow stepped on or sat back on it, the feces were scraped off with a long hoe and placed in a plastic tray. After measuring the weight of the tray on which the collected fecal samples were placed, 100-150 g of samples that could be representative among fecal samples were retaken every 3 hours. Fecal samples collected daily during the set period were stored at -20 ⁰ C until chemical composition analysis. The urine collection method was as follows: A Foley catheter (24 French, 75-cc balloon; CR Bard, Covington, GA) was connected to an approximately 2-3 m long Tygon tube (Fisher Scientific, Waltham, MA) and then disinfected. Placed in a plastic bottle (Fisher Scientific, Waltham, MA). A urine catheter was placed immediately between the time the cattle moved to the metabolic stalls (8 am to 10 am). Samples were taken after allowing 24 hours to acclimate. After catheterization, the cow's condition was somewhat stable. All collected urine samples were weighed, and after sampling, the container containing urine was emptied every 3 hours. 5 ml of urine was placed in a plastic container containing 180 ml of sulfuric acid (concentration 0.5 mol/L), followed by pipetting. The oxidized urine sample was used for nitrogen concentration analysis and stored at -20 ^o C. The following analyzes were performed on feed ingredients and feed residue samples: DM (135 ^o C, AOAC 2000; method 930.15), CP (6.25 Х Kjeldahl N, AOAC 2000; method 990.03), NDF (Van Soest et al. , 1991), ADF (AOAC, 2000; method 973.18), lignin (Goering and Van Soest, 1970), starch starch (Hall, 2009 with correction for free glucose), total ash (535 ^o C, AOAC, 2000) ; method 942.05) and individual mineral content (AOAC, 2000). After drying and grinding the fecal sample (1.0 mm screen), specific chemical components were analyzed.

As a statistical analysis, a linear complex model R (version 3.1.1, R) to find out what effect beta metase was added to feed on feed conversion efficiency (FCE), DMI, BUN and somatic cell count Foundation for Statistical Computing, Vienna, Austria).

Experimental Example 1: Dry matter intake and milk component

As shown in Table 3 below, protein reduction and beta-met nanase addition in feed did not affect dry matter intake. However, cattle ingesting dry matter (LCPE1) with a low crude protein mixed with 0.1% beta metase (LCPE1) had a lower dry matter intake rate than those fed high crude protein (HCP) ( P = 0.068). In light of the results of past experiments, the results of dry matter intake (DMI) were jagged when beta-met nanase was administered to ruminants. For example, in the past, no change in dry matter intake (DMI) was found in cows given beta-met kinase, or 0.1% beta-met kinase was mixed with dry matter and given to cows (0.2% is not the case). It was reported that the dry matter intake was reduced by 1.8 kg.

	Experimental group
	HCP	LCP	LCPE1	LCPE2	LCPE3	SEM
Intake (kg/d)	26.6	25.8	25.0	25.1	24.9	0.81
Yield (kg/d)
milk	37.7	37	39.2	36.1	35.4	1.55
butterfat	1.15	1.21	1.28	1.22	1.12	0.08
Milk protein	1.22	1.17	1.23	1.19	1.05	0.05
Lactose (lactose, lactose)	1.8	1.69	1.80	1.71	2.11	0.1
Milk solids	3.36	3.21	3.38	3.16	3.04	0.16
Milk Ingredients,
Fat, %	3.15	3.23	3.32	3.30	3.28	0.13
protein, %	3.23	3.17	3.16	3.12	2.98	0.07
Lactose (lactose),%	4.68	4.57	4.61	4.71	5.25	0.09
Non-fat milk solids,%	8.8	8.65	8.69	8.52	8.12	0.13
Milk urea nitrogen compound (mg/dL)	14.7	13.0	12.4	12.9	13.1	0.47
Plasma urea nitrogen compound (mg/dL)	18.1	16.7	15.6	16.1	16.5	0.44
Somatic cell count (Х10 3 /mL)	97	111	59	61	65	16.1

In the relative flow rate, it was found that the specific flow rate was increased in the group ( P = 0.09) that consumed beta met kinase (LCPE1) than the cows that consumed low crude protein (LCP). In addition, feed (LCPE2) with a lower soybean meal content as a protein source and a higher Distillers grain content (LCPE2), and a lower soybean meal content as a protein source and steam-flaked corn as a carbohydrate source. When using the diet with high content (LCPE3), the relative milk content was lower than that of the group that consumed the beta metase (LCPE1). In the case of the composition in milk, the low crude protein feed (LCP) and the low crude protein supplemented with beta metase enzyme were added. In the case of mixed feed (LCPE1) and feed with high distillers grain (LCPE2), the composition of milk was not significantly changed, but feed with increased steam-flaked corn content (LCPE3), a source of carbohydrates. There were changes such as a large increase in lactose and a large decrease in protein.

In the case of nitrogen compounds, lower milk urea nitrogen (MUN) was found in the group that consumed lower crude protein (LCP) or lower crude protein and beta met kinase (LCPE1) than the group that consumed high crude protein (HCP). In the past, it has been argued that protein in feed is one of the factors that determine MUN levels. In our experiment, when comparing MUN levels in the two groups ingesting low crude protein, no difference was found between the two groups. As can be seen in Table 3, the plasma urea nitrogen (plasma urea N) value of the group ingesting high crude protein (HCP) was higher than that of the group administered with low crude protein (LCP) and low crude protein and beta metase (LCPE1). This is consistent with the fact that high intake of nitrogen increases MUN levels. The group receiving low crude protein plus beta met kinase (LCPE1) had lower plasma urea nitrogen values ( P = 0.051) than the group receiving the low crude protein (LCP). The reason for this is probably because the group who received beta metase on low crude protein had more nitrogen in milk.

The number of somatic cells in milk was significantly lower in the group receiving low crude protein plus beta metase (LCPE1) than the group receiving low crude protein (LCP, P = 0.0038) or high crude protein (HCP, P = 0.045). In addition, the somatic cell count was also low in the feed with increased Distillers grain content (LCPE2) and the feed with increased steam-flaked corn content (LCPE3), a carbohydrate source.

Experimental Example 2: Total digestive absorption and feed conversion efficiency (FCE)

Table 4 below shows the total average digestive absorption rate values for DM, OM, ADF, NDF, CP, starch, and ash. There was no effect on DM, OM and nutrient absorption rates. These results are similar to those of previously known beta-met kinase experiments. Similarly, there was no difference in DMI and nutrient digestibility, which is believed to be due to no significant difference in OM absorption as a result (15.7 vs. 15.3 vs 16.0 kg/d in HCP, LCP and LCPE1 experimental groups). However, there was a difference in the absorption rate of CP according to the various CP levels contained in the diet (AACP = 2.72 vs. 2.39 kg/d when comparing the HCP subgroup with LCPE, P = 0.035; Table 3). However, compared with the HCP group, there was a tendency for the AACP to decrease in the LCP group (2.72 vs. 2.46 kg/d,; P = 0.1), and there was no difference between the LCP and LCPE1 groups (2.46 vs. 2.39, P). = 0.89).

	Experimental group
	HCP	LCP	LCPE1	LCPE2	LCPE3	SEM
Digestibility (%)
Building (DM)	61.0	62.4	61.4	61.3	61.9	1.27
OM	63.3	63.4	64.5	64.1	63.9	1.31
ADF	42.1	39.2	35.5	36.1	35.8	2.52
NDF	43.7	41.2	42.4	42.1	41.8	1.91
ashes	32.9	33.4	34.3	34.5	34.9	3.2
Crude protein	64.0	63.6	63.3	63.1	62.5	1.29
carbohydrate	96.1	96.9	96.7	96.5	98.1	0.3
AAOM, kg/d	15.7	15.3	16.0	15.8	17.1	0.71
AACP, kg/d	2.72	2.46	2.39	2.40	2.21	0.11
efficiency
MY:DMI	1.43	1.46	1.62	1.51	1.52	0.05
MY:AAOM	2.61	2.47	2.77	2.62	2.53	0.081
Milk protein: CP intake	0.30	0.32	0.34	0.33	0.29	0.014
Milk Protein: AACP	0.48	0.51	0.56	0.52	0.49	0.031

^* AAOM = apparently absorbed organic matter, AACP = apparently absorbed crude protein

Feed conversion efficiency was calculated as the flow rate: dried fish ratio (Table 4). The values calculated to effectively convert CP and ACCP in feed to milk protein are shown in Table 4 above. Additionally, the value at which the absorbed OM effectively converts into flow rate was also calculated.

The feed conversion efficiency (FCE) was increased by 190g per kg of DMI in the group fed LCPE1 than the small group fed HCP feed (13.4% improvement, P = 0.003). When compared with the experimental group that consumed LCP feed, the feed conversion efficiency (FCE) of the LCPE1 group was significantly increased ( P = 0.014) (160g per kg of DMI; 11.0% improvement). Therefore, as a causative factor for the increase in FCE, not only the decreased CP level but also the addition of beta met kinase plays a role. In the case of feed with increased Distillers grain content (LCPE2) and feed with increased steam-flaked corn content (LCPE3), which is a source of carbohydrates, feed conversion efficiency (FCE) was found in the group that consumed HCP feed and It was higher than the LCP diet group, but lower than the LCPE1 group.

Whether or not absorbed OM was effectively used in the body was not known exactly, but OM was certainly used effectively in the LCPE1 experimental group ( P = 0.1). The LCPE1 group converted CP to milk protein more efficiently than the HCP group (milk protein: CP intake = 0.34 vs. 0.30).

Due to incomplete digestion of intestinal proteins, there are limitations in reducing nitrogen levels and synthesizing intestinal nucleic acids and maintaining animals. In order to quantitatively reduce nitrogen in feed, it is necessary to provide nitrogen that can be degraded in the intestine or to allow the absorbed amino acids to be efficiently converted into the synthesis of milk production. In this experiment, the inventors added an enzyme to increase the nitrogen efficiency in milk, which played an important role in converting AACP into milk protein (0.48 vs 0.56 HCP vs LCPE1, P = 0.017, Table 4).

The LCPE1 experimental group had better efficiency of converting AACP to milk protein than the LCP experimental group ( P = 0.1). The reason why cows fed LCP and LCPE1 diets are so efficient is that CP loss can reduce the energy consumption required to release excess nitrogen by providing only the right amount of protein. Thus, the LCPE1 and LCP experimental groups were able to reserve more energy for milk production than the HCP experimental group.

Experimental Example 3: Nitrogen distribution and phosphorus emission

Table 5 lists nitrogen intake, nitrogen excreted in urine and feces, nitrogen excreted in milk, and the amount of phosphorus excreted in manure.

Item	Experimental group
	HCP	LCP	LCPE1	LCPE2	LCPE3	SEM
N intake (g/d)	664	609	598	601	588	23.6
Fecal N (g/d)	239	218	216	217	211	9.6
N output in urine (g/d)	229	192	181	179	176	6.37
Fecal N + urine N (g/d)	442	388	377	376	371	11.6
Milk protein N + MUN (g/d)	207	199	205	204	201	8.1
Preserved/retained N (g/d)	15	22.3	16	18	19	17
Manure P (g/d)	85	82	80	81	79	3.4

Without the effect of DMI, the total amount of nitrogen in the experimental group fed LCP and LCPE1 was decreased compared to the experimental group fed the HCP feed.

Similarly, there was no effect on CP digestibility, and the amount of nitrogen excreted through feces decreased with decreased CP intake. In the experimental group whose CP level was lower than that of the HCP group, the level of nitrogen excreted in the urine decreased. Reducing the amount of nitrogen excreted in the urine is important because the nitrogen excreted in the urine is more likely to be converted to volatile nitrogen than the nitrogen excreted in the feces.

On average, 25% nitrogen excreted in manure, 3 to 15% nitrogen excreted in urine, or 4 to 52% nitrogen excreted in the urine, depends on soil type, moisture, temperature, wind speed, and the concentration of nitrogen in urine and the composition of urine. Therefore, it is discharged in the form of ammonia. Since urine acts as a major factor in the excretion of nitrogen, it is important to control the excretion pathway of nitrogen in order to reduce nitrogen dioxide. Nitrogen dioxide (which emits greenhouse gases that causes global warming) is 265 times more dangerous than carbon dioxide.

The milk of the LCP and LCPE1 experimental groups contained more nitrogen than the HCP. However, the addition of beta met kinase did not have any effect on the total amount of nitrogen discharged as manure (LCP vs LCPE1 experimental group; Table 5).

The most effective way to reduce nitrogen emissions while reducing the environmental impact of nitrogen is to reduce the CP content in the feed. It has already been demonstrated in several papers that the higher the amount of nitrogen consumed as feed, the higher the amount of total nitrogen excreted in urine and feces. It is inefficient if CP is added to feed at a higher level than necessary. The reason for this is that first, not only urea synthesis energy is required to discharge urea into urine, but also more nitrogen than necessary is discharged into the feed as manure, making manure expensive from the perspective of the producer. As shown in our experiment, if the intake of nitrogen is reduced, the amount of nitrogen excreted outside the body is naturally reduced, which can increase the nitrogen use efficiency. As cow's milk production increases, the efficiency of nitrogen use increases and the amount of nitrogen discharged relative to milk production decreases at the same time. However, reducing the amount of feed nitrogen below the appropriate level required in the bovine body can adversely affect productivity. In this experiment, while reducing the CP level in the feed, the milk production level was maintained by adding a beta metase.

In conclusion, by reducing the CP level in the feed and adding beta metase, the cost can be reduced from the perspective of feed producers. Using the California 2018 feed standard to calculate the cost, the money needed to produce the same amount of milk is: HCP = $9.41, LCP = $8.65 and LCPE1 = $8.38 (feed per cow). Therefore, if beta metase was added instead of reducing the CP level in the feed, it would save $1.03 per cow per day in lactating cows.

In the above, the present invention has been illustrated and described in relation to specific preferred embodiments, but the present invention may be variously modified and changed within the scope of not departing from the technical features or field of the present invention provided by the following claims. It is obvious to one of ordinary skill in the art that it can be.

Claims (12)

1. Corn silage (Corn silage), steam-flaked corn (Steam-flaked corn), and one or more carbohydrate sources selected from the group consisting of flat barley rice (Rolled barley);

   A protein source including soybean meal, and further comprising one or more selected from the group consisting of Alfalfa hay, Distillers grain, and Soybean hulls;

   A local source of cotton seeds; And

   Consists of including beta-mannanase (β-mannanase) as an enzyme,

   The low-protein feed composition for enhancing feed conversion efficiency (FCE) of ruminants, wherein the soybean meal is contained within the range of 4.0 to 7.0 parts by weight based on 0.1 parts by weight of the beta met nanase.
2. The method of claim 1,

   Crude protein (CP: Crude Protein) is characterized in that contained within the range of 14.5 to 15.0% by weight, low protein feed composition for improving feed conversion efficiency of ruminants.
3. The method of claim 2,

   The low-protein feed composition for improving feed conversion efficiency of ruminants, characterized in that the soybean meal is contained within the range of 5.0 to 5.5 parts by weight based on 0.1 parts by weight of the beta met nanase.
4. The method of claim 3,

   The carbohydrate source is characterized in that consisting of corn silage 25.0 to 26.0 parts by weight, steam-flaked corn 11.0 to 12.0 parts by weight, and rolled barley 7.0 to 8.0 parts by weight, Low protein feed composition for enhancing the feed conversion efficiency of ruminants.
5. The method of claim 4,

   The protein source is 4.0 to 7.0 parts by weight of Soybean meal, 26.0 to 27.0 parts by weight of Alfalfa hay, 5.0 to 6.0 parts by weight of Distillers grain, and 11.0 to 12.0 parts by weight of Soybean hulls A low-protein feed composition for enhancing feed conversion efficiency of ruminants, characterized in that the addition is included.
6. The method of claim 5,

   The fat source is a low-protein feed composition for improving feed conversion efficiency of ruminants, characterized in that consisting of 6.0 to 7.0 parts by weight of cotton seed (Cotton seed).
7. The method according to any one of claims 1 to 6,

   A low protein feed for enhancing feed conversion efficiency of ruminants, characterized in that it further comprises at least one selected from the group consisting of mineral mix, calcium carbonate, and salt (NaCl) Composition.
8. The method of claim 7,

   The mineral mix (Mineral mix) 1.0 to 2.0 parts by weight, calcium carbonate (Calcium carbonate) 0.1 to 0.2 parts by weight, and salt (NaCl), characterized in that it further comprises 0.1 to 0.2 parts by weight, ruminant feed Low protein feed composition for improving conversion efficiency.
9. The method according to any one of claims 1 to 6,

   Compared with a feed composition that does not contain beta-mannanase as an enzyme, feed conversion efficiency is increased, and urea mass (MUN) and somatic cell count are lowered, thereby enhancing feed conversion efficiency of ruminants. For low protein feed composition.
10. Corn silage 25.0 to 26.0 parts by weight, steam-flaked corn 11.0 to 12.0 parts by weight, and a carbohydrate source consisting of 7.0 to 8.0 parts by weight of rolled barley;

    Soybean meal 4.0 to 7.0 parts by weight, Alfalfa hay 26.0 to 27.0 parts by weight, Distillers grain 5.0 to 6.0 parts by weight, and Soybean hulls 11.0 to 12.0 parts by weight. Protein source;

    Fat source consisting of 6.0 to 7.0 parts by weight of cotton seed; And

    As an enzyme, beta met nanase (β-mannanase) 0.1 parts by weight; consisting of, including, feed conversion efficiency (FCE: Feed Conversion Efficinecy) of a ruminant low-protein feed composition for enhancing.
11. The method of claim 10,

    Mineral mix (Mineral mix) 1.0 to 2.0 parts by weight, calcium carbonate (Calcium carbonate) 0.1 to 0.2 parts by weight, and salt (NaCl) 0.1 to 0.2 parts by weight, characterized in that it further comprises a ruminant feed Low protein feed composition for improving conversion efficiency.
12. Corn silage (Corn silage) 25.65 parts by weight, steam-flaked flakes (Steam-flaked corn) 11.3 parts by weight, and a carbohydrate source consisting of 7.21 parts by weight of rolled barley;

    A protein source comprising 5.21 parts by weight of Soybean meal, 26.2 parts by weight of Alfalfa hay, 5.01 parts by weight of Distillers grain, and 11.43 parts by weight of Soybean hulls;

    Fat source consisting of 6.55 parts by weight of cotton seed;

    0.1 parts by weight of beta-mannanase as an enzyme;

    1.09 parts by weight of mineral mix;

    0.16 parts by weight of calcium carbonate; And

    A low-protein feed composition for improving feed conversion efficiency of ruminants, comprising 0.19 parts by weight of salt (NaCl).