Composition for inhibiting bile salt hydrolase comprising epicatechin and use thereof

Abstract

This invention relates to: a composition for inhibiting bile salt hydrolase (BSH) comprising epicatechin as an active ingredient; a method for inhibiting bile salt hydrolase in animals using the composition; and the use of epicatechin in inhibiting bile salt hydrolase.

Description

[Technical Field]

[0001] This invention relates to: a composition for inhibiting bile salt hydrolase (BSH), comprising epicatechin as an active ingredient; a method for inhibiting bile salt hydrolase in animals using the composition; and the use of epicatechin in inhibiting bile salt hydrolase. [Previous Technology]

[0002] Bile salt hydrolase is an enzyme produced by intestinal microorganisms as a self-defense mechanism against bile acids. It has been reported that inhibiting bile salt hydrolase can improve the growth performance of livestock (Begley, M.; Hill, C.; Gahan, CGM (2006) Bile Salt Hydrolase Activity in Probiotics. Applied and Environmental Microbiology, 72(3), 1729-1738).

[0003] Although some compounds have been studied as inhibitors of bile salt hydrolase, it is well known that such synthetic compounds may cause side effects in livestock. Therefore, there is a need to develop substances that can inhibit bile salt hydrolase while ensuring safety. [Summary of the Invention]

[0004] [Technical Issues]

[0005] The purpose of this invention is to provide a composition for inhibiting bile salt hydrolytic enzymes, which contains epicatechin as an active ingredient.

[0006] Another object of the present invention is to provide a method for inhibiting bile salt hydrolases in animals, comprising feeding the animal epicatechin.

[0007] Another object of the present invention is to provide a method for increasing the weight of an animal, comprising feeding the animal epicatechin.

[0008] Another object of the present invention is to provide the use of epicatechin in inhibiting bile salt hydrolase.

[0009] Another object of the present invention is to provide the use of epicatechin in increasing the weight of animals.

[0010] [Technical Solution]

[0011] One aspect of the present invention provides a composition for inhibiting bile salt hydrolase, comprising epicatechin as an active ingredient.

[0012] Another aspect of the present invention provides a method for inhibiting bile salt hydrolases in an animal, comprising feeding the animal epicatechin.

[0013] Another aspect of the present invention provides a method for increasing the weight of an animal, comprising feeding the animal epicatechin.

[0014] Another aspect of the present invention provides the use of epicatechin in inhibiting bile salt hydrolase.

[0015] Another aspect of the present invention provides the use of epicatechin in increasing the weight of animals.

[0016] [Beneficial Effects]

[0017] This invention helps to inhibit bile salt hydrolytic enzymes in animals, promote livestock growth, improve feed efficiency, and enhance feed quality.

Implementation Method

[0018] The present invention will be described in detail below. Furthermore, each description and embodiment described herein can be applied to other descriptions and embodiments. That is, all combinations of the various elements described herein fall within the scope of the present invention. Moreover, the scope of the present invention is not limited to the specific descriptions below.

[0019] Furthermore, those skilled in the art can identify or confirm many equivalents of the specific aspects of the invention described herein through conventional experiments. Moreover, it is intended that such equivalents be included in the present invention.

[0020] As used in the specification and appended claims of this invention, unless the context clearly indicates otherwise, the singular articles (“a,” “an,” and “the”) include the plural referents. Furthermore, unless the context clearly indicates otherwise, singular terms include their plural forms, and plural terms include their singular forms. As used in the specification and appended claims of this invention, unless otherwise indicated, the term “or” may include the meaning of “and / or”.

[0021] As used herein, the term "about" may precede a specific numerical value. As used herein, the term "about" encompasses not only the exact specified numerical value but also a range approximately or close to that value. Considering the context in which the numerical value appears, it can be determined that the term refers to a value approximately or close to the specified value. For example, the term "about" may indicate a range of -10% to +10% of the value. As another example, the term "about" may indicate a range of -5% to +5% of the value, but is not limited to this.

[0022] As used herein, terms such as "first," "second," "third," "i," "ii," "iii," or "(a), (b), (c), (d)" are used to distinguish similar elements and do not imply that these elements are performed consecutively or in the listed order. For example, when these terms are used in relation to methods, uses, or analytical steps, these steps may be performed without any time interval, simultaneously, or at intervals of seconds, minutes, hours, days, or months.

[0023] As used herein, the term "comprising" means the features, steps, or components that follow the term, and does not exclude the presence or addition of one or more other features, steps, or components. As used herein, the components or features following the term "comprising" may be essential or mandatory; however, in some embodiments, other optional or non-essential components or features may also be included.

[0024] One aspect of the present invention provides a composition for inhibiting bile salt hydrolase (BSH), which includes epicatechin as an active ingredient.

[0025] As used herein, the term "catechin" refers to a form of polyphenol and may include one or more selected from the group consisting of: epigallocatechin (EGC), epicatechin (EC), epigallocatechin gallate (EGCG), epicatechin gallate (ECG), gallatechin (GC), catechin gallate (CG), gallatechin gallate (GCG), and catechin (C). In particular, catechins are classified as epicatechin and non-epicatechin, and the catechin of the present invention may be epicatechin.

[0026] The epicatechin of the present invention may be EGCG, EGC, or a combination thereof.

[0027] In one embodiment, the epicatechin of the present invention (specifically EGCG, EGC, or a combination thereof) may be included in the composition for inhibiting bile salt hydrolase in the following amounts: 1 ppm or more, specifically 1 ppm to 1000 ppm, 1 ppm to 500 ppm, 1 ppm to 125 ppm, 1 ppm to 62 ppm, 1 ppm to 31 ppm, 2 ppm to 1000 ppm, 2 ppm to 500 ppm, 2 ppm to 125 ppm, 2 ppm to 62 ppm, 2 ppm to 31 ppm, 31 ppm to 1000 ppm, 31 ppm to 500 ppm, 31 ppm to 125 ppm, 31 ppm to 62 ppm, 62 ppm to 1000 ppm, 62 ppm to 500 ppm, 62 ppm to 125 ppm, 125 ppm to 1000 ppm. ppm, or 125 ppm to 500 ppm, but not limited to these.

[0028] In one embodiment, epicatechin may exhibit bile salt hydrolase inhibitory activity.

[0029] As used herein, the term “bile salt hydrolase (BSH)” refers to an enzyme whose primary function is to convert conjugated bile salts into deconjugated bile salts. Bile salt hydrolases can be produced by, but are not limited to, intestinal commensal bacteria. In addition, antibiotic growth promoters (AGPs) are commonly used to promote livestock growth; however, due to the problems arising from the abuse and overuse of antibiotics in livestock, antibiotics need to be used with caution and regulated. Some antibiotics used in feed exhibit bile salt hydrolase inhibitory activity, reducing the activity of bile salt hydrolase and thus promoting animal growth, and bile salt hydrolase inhibitors have been reported as alternatives to such antibiotics (Begley, M.; Hill, C.; Gahan, CGM (2006). Bile Salt Hydrolase Activity in Probiotics. Applied and Environmental Microbiology, 72(3), 1729-1738). In this regard, the epicatechin of the present invention exhibits excellent effects in improving feed efficiency by inhibiting bile salt hydrolase and increasing animal weight.

[0030] In one embodiment, the epicatechin EGCG and EGC of the present invention exhibit superior bile salt hydrolase inhibitory activity compared to epicatechin gallate (ECG).

[0031] In one embodiment, the bile salt hydrolase of the present invention can be derived from one or more microorganisms selected from the group consisting of Bacteroides, Escherichia, and Salmonella, and specifically from one or more microorganisms selected from the group consisting of Bacteroides fragilis, Escherichia coli, Salmonella typhimurium, and Salmonella infantis, but is not limited thereto. Escherichia coli can be Shiga toxin-producing Escherichia coli stx2e and / or pathogenic Escherichia coli APEC O, but is not limited thereto.

[0032] The composition of the present invention for inhibiting bile salt hydrolase can be used to increase animal weight. It has been reported that bile salt hydrolase can promote animal growth (Begley, M.; Hill, C.; Gahan, CGM, (2006). Bile Salt Hydrolase Activity in Probiotics. Applied and Environmental Microbiology, 72(3), 1729-1738), and the present invention first discovered that epicatechin of the present invention exhibits excellent bile salt hydrolase inhibitory efficacy. Therefore, the composition of the present invention can be used to increase animal weight.

[0033] As used herein, the terms "inhibiting bile salt hydrolase" or "bile salt hydrolase inhibition" refer to the inhibition of bile salt hydrolase activity.

[0034] In one embodiment, bile salt hydrolase inhibition may include a decrease or inactivation of the activity of the bile salt hydrolase in the animal compared to its intrinsic activity prior to feeding the animal with the feed, feed composition, feed additive, and / or feed additive composition of the present invention. In other words, inhibition may include both: decreased activity compared to the intrinsic activity of the enzyme prior to feeding the animal with the feed, feed composition, feed additive, and / or feed additive composition of the present invention; or complete inactivation.

[0035] The term "intrinsic activity" refers to the bile salt hydrolytic enzyme activity that an animal originally possesses before the feed, feed composition, feed additive, and / or feed additive composition of the present invention are fed to the animal.

[0036] In addition, the enzyme activity of the present invention can be evaluated by the conversion rate of the enzyme matrix (e.g., bile salts) to the product, the degree of matrix reduction when the enzyme is added to the matrix, and / or the degree of matrix reduction compared to the initial amount administered or fed to the animal, but is not limited thereto.

[0037] In one embodiment, the inhibition of activity may refer to a reduction in the function of converting bound bile salts to unbound bile salts, and when quantified according to the formula "%BSH inhibition rate = (OD material treated - OD strain) / (OD strain / matrix - OD strain) × 100 (%)", it may have a positive (+) value (where OD material treated = absorbance after reaction of sample, strain and matrix; OD strain = absorbance after treatment of strain alone; and OD strain / matrix = absorbance after reaction of strain and matrix).

[0038] One embodiment of the present invention provides a feed or feed additive containing epicatechin as an active ingredient. The epicatechin may be EGCG, EGC, or a combination thereof, and may exhibit bile salt hydrolase inhibition.

[0039] The composition of the present invention for inhibiting bile salt hydrolase can be used as feed. Therefore, the composition of the present invention for inhibiting bile salt hydrolase can be a feed composition for inhibiting bile salt hydrolase, and / or a feed additive composition.

[0040] As used herein, the term "feed" (including "feed composition" and "feed additive") refers to food fed to animals. Feed may include substances that provide organic or inorganic nutrients necessary for maintaining animal life or for producing meat, dairy products, etc. Feed may further contain other nutrients necessary for maintaining animal life or for producing meat, dairy products, etc. The feed of the present invention may contain the composition of the present invention for inhibiting bile salt hydrolysates, the feed composition of the present invention, and the feed additive of the present invention.

[0041] As used herein, the term "feed additive" refers to a substance added to a feed composition and which may be considered a supplement to feed under feed regulations. Feed additives may be used to improve the productivity or health of animals or livestock, but are not limited thereto. The feed additives of the present invention may include the components of the present invention that inhibit bile salt hydrolysates, or the feed additive components of the present invention.

[0042] The feed, feed composition, feed additive, and / or feed additive composition of the present invention can be used on animals, but are not limited thereto, as long as the species can be fed with feed.

[0043] The feed, feed composition, feed additive, and / or feed additive composition of the present invention can be used in animal diets. Humans are excluded, but there are no particular limitations. The animal can include a variety of animals, including mammals and poultry. The feed can be used on mammals such as pigs, cattle, sheep, goats, laboratory rodents, and companion animals (e.g., dogs and cats), and on poultry such as chickens, turkeys, ducks, geese, pheasants, and quails. The method of feeding the feed is not limited.

[0044] The type of feed or feed additive of the present invention is not particularly limited, and feeds commonly used in the art can be used. Non-limiting examples of feeds or feed additives include: plant-based feeds or feed additives, such as cereals, root crops, food processing by-products, algae, fibrous materials, pharmaceutical by-products, fats and oils, starch, gourd or cereal by-products; and animal-based feeds or feed additives, such as proteins, minerals, fats and oils, mineral-based substances, single-cell proteins, animal plankton, or food. These examples can be used alone or in combination of two or more.

[0045] The feed, feed composition, feed additive, and / or feed additive composition of the present invention can be manufactured in powder or granule form, and, if necessary, may further include one or more of the following: organic acids, such as citric acid, fumaric acid, adipic acid, lactic acid, or malic acid; phosphates, such as sodium phosphate, potassium phosphate, acid pyrophosphate, or polyphosphate; or natural antioxidants, such as polyphenols, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and pectin acid. The feed or feed additive of the present invention can be formulated in the form of conventional feed and may further include conventional feed ingredients.

[0046] The feed, feed composition, feed additive, and / or feed additive composition of the present invention may further include: cereals such as ground or crushed wheat, oats, barley, corn, and rice; plant proteins such as preparations or substances with rapeseed, soybean, and sunflower as the main components; animal proteins such as blood meal, meat meal, bone meal, and fish meal; and carbohydrates and dairy products such as various dried components composed of milk powder and whey powder, and may also include additional supplements, digestion and absorption enhancers, growth promoters, etc.

[0047] The feed, feed composition, feed additive, and / or feed additive composition of the present invention can be administered to animals alone or in combination with other feed additives in an edible carrier. Furthermore, the feed, feed composition, feed additive, and / or feed additive composition of the present invention can be readily administered to animals as a top dressing by directly mixing it into feed or as a separate oral dosage form. When administered, the feed, feed composition, feed additive, and / or feed additive composition of the present invention can be formulated as an immediate-release or sustained-release formulation by combining it with a pharmaceutically acceptable edible carrier, as is known in the art. The edible carrier can be solid or liquid and may include, for example, corn starch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil, and propylene glycol. If a solid carrier is used, feed additives may be in the following forms as top dressing: tablets, capsules, powders, troches, sugar-coated tablets, or non-dispersible forms. If a liquid carrier is used, feed additives may be in the form of soft gelatin capsules, syrups, suspensions, emulsions, or solutions.

[0048] The feed, feed composition, feed additive, and / or feed additive composition of the present invention may contain, for example, preservatives, stabilizing agents, moisturizing agents, emulsifiers, or solution promoters. The feed additive, or feed additive composition, may be added, sprayed, or mixed into animal feed for use.

[0049] Regarding application, the feed, feed composition, feed additive, and / or feed additive composition of the present invention may be further mixed with one or more of the following groups: organic acids such as citric acid, fumaric acid, adipic acid, and lactic acid; phosphates such as potassium phosphate, sodium phosphate, and polyphosphates; and natural antioxidants such as polyphenols, catechins, tocopherols, vitamin C, green tea extract, chitosan, and tannic acid. Other known additives, such as anti-influenza agents, buffers, and antibacterial agents, may be added if necessary. Furthermore, the feed, feed composition, feed additive, and / or feed additive composition of the present invention may be formulated into injectable forms such as solutions, suspensions, or emulsions, as well as capsules, granules, or tablets, by adding additional diluents, dispersants, surfactants, binders, or lubricants. Furthermore, the feeds, feed compositions, feed additives, and / or feed additive compositions of the present invention can be used in combination with supplementary ingredients, including various supplements such as amino acids, inorganic salts, vitamins, antioxidants, antifungal agents, and antibacterial agents, as well as plant-based protein feeds such as crushed or ground wheat, barley, and corn, and animal-based protein feeds such as blood meal, meat meal, and fish meal. In addition to major components such as animal fats and plant fats, it can be used in combination with nutritional supplements, growth promoters, digestion and absorption enhancers, and disease prevention agents.

[0050] The feed, feed composition, feed additive, and / or feed additive composition of the present invention can be mixed into animal feed based on the dry weight of livestock feed, in an amount of about 10 to 500 grams per kilogram, for example, 10 to 100 grams per kilogram, and can be supplied as a paste after complete mixing, or can be granulated, expanded, or extruded through additional process steps.

[0051] The epicatechin content of the feed, feed composition, feed additive, and / or feed additive composition of the present invention can be appropriately adjusted according to the type and age of the livestock to which it is applied, the form of application, the desired effect, etc., and can be, for example, from 0.01 wt% to 90 wt%.

[0052] Furthermore, the feed, feed composition, feed additive, and / or feed additive composition according to the present invention can be used alone or in combination with conventional feeds or feed additives, and can be used sequentially or simultaneously. Moreover, it can be administered in a single dose or multiple doses. Importantly, while taking all the foregoing requirements into account, the minimum effective amount that maximizes efficacy without causing side effects is determined, and this amount can be readily determined by those skilled in the art.

[0053] Another aspect of the present invention provides a method for inhibiting bile salt hydrolases in an animal, comprising feeding the animal epicatechin.

[0054] In one embodiment, the method may involve increasing the weight of the animal.

[0055] Another aspect of the present invention provides a method for increasing the weight of an animal, comprising feeding the animal epicatechin.

[0056] There are no restrictions on the feeding method, as long as the method is in a form that the animal can consume.

[0057] Another aspect of the present invention provides a method for inhibiting bile salt hydrolase, comprising treating epicatechin to bile salt hydrolase-producing microorganisms.

[0058] Epicatechin, animal and bile salt hydrolytic enzyme systems as described in other examples, and epicatechin may be EGCG, EGC, or a combination thereof.

[0059] A method for inhibiting bile salt hydrolase in animals or for increasing animal weight may specifically include the following steps: (a) mixing a feed, feed composition, feed additive, and / or feed additive composition containing epicatechin as an active ingredient with an animal feed; and (b) feeding the animal feed to the animal.

[0060] Step (a) according to the present invention comprises mixing the feed, feed composition, feed additive, and / or feed additive composition of the present invention, which contains epicatechin as an active ingredient, with general feed for livestock. The mixed feed may be fed containing 0.01 wt% to 90 wt%, and preferably 0.1 wt% to 10 wt% of the feed, feed composition, feed additive, and / or feed additive composition of the present invention.

[0061] Step (b) according to the present invention includes administering the feed containing epicatechin produced in step (a) to an animal, and the method of feeding is not limited. As mentioned above, there are no particular limitations on the animals to be fed, and the feed can be used in the diets of a variety of animals, including mammals and poultry. The feed can be used for mammals such as pigs, cattle, sheep, goats, and laboratory rodents, as well as companion animals (e.g., dogs and cats). It can also be used for poultry such as chickens, turkeys, ducks, geese, pheasants, and quails.

[0062] It is anticipated that feeding animals with the feed, feed composition, feed additive, and / or feed additive composition according to the present invention will exhibit excellent effects in inhibiting bile salt hydrolase (BSH) activity to replace antibiotics, while promoting livestock growth and improving feed efficiency.

[0063] Another aspect of the present invention provides the use of epicatechin, the feed of the present invention, the feed composition of the present invention, the feed additive of the present invention, and / or the feed additive composition of the present invention in inhibiting bile salt hydrolase and promoting weight gain in animals.

[0064] The epicatechin, feed composition, feed additives, and bile salt hydrolytic enzyme system are as described above, and the epicatechin may be EGCG, EGC, or a combination thereof.

[0065] [Methods for implementing the present invention]

[0066] The present invention will be described in detail by way of examples. However, these examples are for illustrative purposes only, and the scope of the invention is not intended to be limited by these examples. Furthermore, any technical descriptions not appearing in the present invention can be fully understood and readily implemented by those skilled in the art or related fields.

[0067] Example 1. BSH inhibitory effect of specific components of epicatechin.

[0068] Example 1-1. Preparation of samples for evaluating BSH inhibitory efficacy

[0069] Liquid culture medium was prepared using LB broth (BD DIFCO 10855021), and Shiga toxin-producing *Escherichia coli* stx2e, *Salmonella typhimurium*, pathogenic *Escherichia coli* APEC O, and *Salmonella infantis* were inoculated into the medium. The mixture was then incubated statically at 37°C for one day. Additionally, *Bacteroides fragilis* was inoculated into enhanced *Clostridium* medium (BD DIFCO 218081) and anaerobic statically incubated at 37°C for two days. In particular, strain information and culture conditions are shown in Table 1 below.

[0070] This culture was prepared by diluting the microbial culture with 0.1 M sodium phosphate buffer (pH 6.0) containing DL-dithiothreitol (DL-DTT) to achieve an optical density of 0.3 (600 nm). The matrix was prepared by dissolving 100 mM glycine deoxycholic acid (GDCA, Sigma G9910) in dimethyl sulfoxide (DMSO, Sigma 276855) and stored in a refrigerator.

[0071] [Table 1] livestock species strain name strain number culture medium Humans Bacteroides fragilis KCTC5013 Enhanced Clostridium culture medium pig Escherichia coli (STEC / Edema Disease) KVCC-BA0001423 LB pig Salmonella typhimurium ATCC14028 LB chicken Pathogenic Escherichia coli (APEC) (O) CR02-A0054 LB chicken Salmonella in infants SARB26 (SGSC2483) LB

[0072] Example 1-2. Evaluation method of BSH inhibitory efficacy

[0073] As evaluation materials, epicatechin gallate (ECG), epigallocatechin gallate (EGCG), epigallocatechin (EGC), and tetracycline were diluted with 0.1 M sodium phosphate buffer containing 5 mM DTT and prepared according to each concentration for evaluation. After reacting the prepared materials with BSH-producing strain cultures at 37°C for 30 minutes, a matrix, glycodeoxycholic acid, was added. The reaction was terminated when the optical density (OD 600 nm) of the reaction mixture containing the matrix and strain reached 0.5 or higher. The % inhibition rate was then calculated as follows to compare the BSH inhibitory efficacy of each material.

[0074] - %BSH inhibition rate = (OD material treated – OD strain) / (OD strain / matrix – OD strain) × 100 (%). In this paper, OD material treated = absorbance of sample, bacteria and matrix after reaction; OD strain = absorbance of bacteria after treatment alone; and OD strain / matrix = absorbance of bacteria and matrix after reaction.

[0075] [Table 2] Classification Test materials information Catechin epicatechin gallate (ECG) Sigma, E3893 Epigallocatechin gallate (EGCG) Sigma, PHR1333 Epigallocatechin gallate (EGC) Sigma, E3768 antibiotic Tetracycline Sigma, O4636

[0076] Examples 1-3. Evaluation results of BSH inhibitory efficacy

[0077] The positive control group, tetracycline, was confirmed to have a 100% inhibition rate of %BSH against Bacteroides fragilis at a concentration of 125 ppm. At 125 ppm, ECG showed a 46% inhibition rate, while EGCG and EGC showed 65% and 73% inhibition rates, respectively, indicating higher inhibition rates compared to ECG. Furthermore, at a concentration of 62 ppm, ECG showed a 31% inhibition rate, while EGCG and EGC showed 48% and 45% inhibition rates, respectively. Moreover, at a concentration of 31 ppm, ECG showed a 9% inhibition rate, while EGCG and EGC showed 15% and 13% inhibition rates, respectively, indicating higher inhibition rates compared to ECG (Table 3).

[0078] [Table 3] Classification Material BSH inhibition efficacy at 125 ppm (%) BSH inhibition efficacy at 62 ppm (%) BSH inhibition efficacy at 31 ppm (%) Catechin EGC 46 31 9 EGCG 65 48 15 EGC 73 45 13 antibiotic Tetracycline 100 Not detected Not detected

[0079] The %BSH inhibition rate of various materials against porcine Escherichia coli edema strains was determined at a concentration of 31 ppm. As shown in the results, ECG, EGCG, and EGC showed inhibition rates of 23.52%, 37.14%, and 52.38%, respectively (Table 4).

[0080] [Table 4] Classification Material BSH inhibition efficacy at 31 ppm (%) Catechin EGC 23.52 EGCG 37.14 EGC 52.38

[0081] The %BSH inhibition rate of various materials against Salmonella typhimurium in pigs was determined at a concentration of 31 ppm. As shown in the results, ECG, EGCG, and EGC showed inhibition rates of 10.00%, 30.23%, and 15.00%, respectively (Table 5).

[0082] [Table 5] Classification Material BSH inhibition efficacy at 31 ppm (%) Catechin EGC 10.00 EGCG 30.23 EGC 15.00

[0083] The %BSH inhibition rate of various materials against avian pathogenic Escherichia coli APEC(0) in chickens was determined at a concentration of 62 ppm. As shown in the results, ECG, EGCG, and EGC showed inhibition rates of 6.23%, 28.82%, and 10.32%, respectively (Table 6).

[0084] [Table 6] Classification Material BSH inhibition efficacy at 62 ppm (%) Catechin EGC 6.23 EGCG 28.82 EGC 10.32

[0085] The %BSH inhibition rate of various materials against Salmonella infantis in chickens was determined at a concentration of 62 ppm. As shown in the results, ECG, EGCG, and EGC showed inhibition rates of 18.43%, 32.32%, and 61.40%, respectively (Table 7).

[0086] [Table 7] Classification Material BSH inhibition efficacy at 62 ppm (%) Catechin EGC 18.43 EGCG 32.32 EGC 61.40

[0087] Therefore, the above examples demonstrate that EGCG and EGC exhibit superior BSH inhibitory activity compared to ECG, regardless of the type of BSH.

[0088] Example 2. BSH-inhibiting effects of natural substances containing epicatechin.

[0089] A liquid culture medium was prepared using LB broth (BD DIFCO 10855021), and *E. coli* (APEC O) was inoculated into the medium. The mixture was then incubated statically at 37°C for two days. The culture was prepared by diluting the microbial culture with 0.1 M sodium phosphate buffer (pH 6.0) containing DL-dithiothreitol (DL-DTT) to achieve an optical density of 0.3 (600 nm). The matrix was prepared by dissolving 100 mM glycine deoxycholic acid (GDCA, Sigma G9910) in dimethyl sulfoxide (DMSO, Sigma 276855) and stored in a refrigerator. The evaluation material, green tea (Boseong, Republic of Korea), was dried to a moisture content of 3% or less, ground, and then treated with 50% methanol for 30 minutes. The resulting extract was used for evaluation.

[0090] It was confirmed that the positive control group, tetracycline, at a concentration of 125 ppm, had a 35% inhibition rate of %BSH produced by avian Escherichia coli APEC O. On the other hand, green tea, a natural substance containing epicatechin, showed a 37% BSH inhibition rate at a concentration of 125 ppm (Table 8).

[0091] [Table 8] Classification Material BSH inhibition efficacy at 125 ppm (%) natural substances green tea 37 antibiotic Tetracycline 35

[0092] Based on the above results, it has been found that epicatechin of the present invention can inhibit bile salt hydrolase, and these results indicate that the present invention can be used as an excellent active ingredient in feed or feed additives.

[0093] As described above, those skilled in the art will understand that the present invention can be implemented in other specific forms without departing from its technical spirit or essential characteristics. Therefore, the above embodiments should be interpreted as illustrative and not as limiting the present invention. It should be understood that all changes or modifications made in accordance with the definition and scope of the claims and their equivalents are within the scope of the present invention.

Claims

1. Use of an epicatechin in the preparation of a medicament for inhibiting bile salt hydrolases, wherein the epicatechin is epigallocatechin (EGC).

2. The use as described in claim 1, wherein the epicatechin further comprises epigallocatechin gallate (EGCG).

3. Use of an epicatechin in the preparation of a medicament for increasing the weight of animals, wherein the epicatechin is epigallocatechin (EGC).

4. The use as described in claim 3, wherein the epicatechin further comprises epigallocatechin gallate (EGCG).