KR20220128106A - Method for producing pet healthy food using fermentation product by Lentinula edodes fermentation of herbs and oak - Google Patents

Abstract

The present invention relates to a method for producing healthy pet food, which comprises a step of adding a fermentation product of Dendropanax morbiferus, Eriobotrya japonica and Avena sativa fermented by Lentinula edodes hypha, specifically, fermented by Lentinula edodes JMIF-001(KCTC18874P) and meat, and extracts of Lentinula edodes, Astragalus membranaceus, Wolfiporia extensa, Phyllostachys nigra and Atractylodes lancea followed by mixing, and to healthy pet food produced by the method. The healthy pet food produced by the method of the present invention has a high content of beta glucan, essential amino acids for pets, vitamins and the like.

Description

{Method for producing pet healthy food using fermentation product by Lentinula edodes fermentation of herbs and oak}

The present invention is after adding water to a mixture of loquat extract powder and hwangchil extract powder, immersing oats here and sterilizing it, and inoculating shiitake mycelium ( Lentinula edodes (Berk.)) JMIF-001 (KCTC18874P) After culturing, drying and pulverizing to prepare a fermented product of hwangchil loquat oat shiitake mycelium; and adding and mixing the prepared hwangchil loquat oat shiitake mycelium fermented product and meat, Wongogogo extract, astragalus extract, bokryeong extract, bamboo leaf extract, and baekchul extract. It relates to a method and a snack for companion animals prepared by the method.

The domestic companion animal market is expected to grow from 2.89 trillion won in 2017 to 5.81 trillion won in 2020. This growth is due to the growing tendency to treat companion animals' food as 'pet food' rather than 'feed' as the perception that companion animals are another family member has become more common.

In line with this trend, large corporations are actively entering the market. Following existing companies such as KGC Ginseng Corporation and Pulmuone, Binggrae also applied for pet food trademark ‘Evergro’ and launched products.

In the United States, the size of the companion animal market in 2015 already exceeded $60 billion (about 67 trillion won), and it is estimated that about 70% of all households have companion animals. A wearable terminal that calculates, monitors, and visualizes the amount of exercise required according to the size and weight of the companion animal, and a matching service that connects the companion animal to the entrusted guardian when the companion animal needs to be entrusted for a while are also emerging. In the UK, about 20% of pet households have pet insurance, and this is a growing trend.

Processed foods using Wongogo mushroom, a representative specialty of Jangheung, and various herbal medicines are currently limited to products for humans. Shiitake mushrooms are rich in vitamin D, which increases the absorption of calcium and helps bone health. Beta glucan also boosts immunity. These effects of shiitake mushrooms are equally applicable to companion animals as well as humans, so they have great value as a raw material for companion animal health snacks. The current trend of companion animal snacks is focused on health functionality beyond palatability. KGC Ginseng Corporation’s ‘Genipet’ using 6-year-old red ginseng is a representative example.

Among the numerous companion animal foods currently circulating on the market, there are many low-grade foods whose safety of raw materials or raw materials is unknown, or chemical components (preservatives, colorants, etc.) are added and nutritionally insufficient. Anxiety about imported raw materials, including China, increased due to the case in the United States, where about 600 dogs died due to junk food from China in 2013. Accordingly, many companies are promoting that they are producing products using domestic raw materials, but consumers still cannot confirm the production or manufacturing process of raw materials.

In the case of 'Chur', a representative snack for cats, the Japanese product 'Chao Chur' has a fairly high market share. Currently, as the trade conflict between Korea and Japan escalates and the boycott of Japanese products intensifies, the demand for wet products manufactured in Korea is increasing. It is necessary to develop competitive products that can replace imported products in line with the trend of increasing consumer needs for domestic products.

Korean Patent Publication No. 2021-0026508 discloses a method for manufacturing a companion animal snack using malt meal, and Korean Patent No. 2099464 discloses a method for manufacturing a companion animal health-improving nutritional snack, but Hwangchil loquat of the present invention It is different from the manufacturing method of healthy snacks for companion animals using fermented oat shiitake mycelium.

The present invention has been derived from the above needs, and in the present invention, oats containing beta-glucan, a useful ingredient related to the immune effect, are used to utilize the ingredients of shiitake mushrooms and herbal medicines that are helpful for various diseases of companion animals, strengthening immunity, etc. By mixing hwangchil and loquat extracts and using the fermented product fermented with shiitake mycelium, which has been patented, the mixed material is used to make snacks for companion animals, it is preferred by companion animals and has excellent nutritional and anti-inflammatory effects. An object of the present invention is to provide a method for preparing snacks.

In order to solve the above problems, the present invention provides a method comprising: (1) roasting loquat, adding water to the pulverized loquat, extracting it, concentrating and freeze-drying to prepare a loquat extract powder; (2) extracting by adding water to the pulverized hwangchil after roasting hwangchil, and concentrating and freeze-drying to prepare hwangchil extract powder; (3) After adding water to a mixture of the loquat extract powder prepared in step (1) and the hwangchil extract powder prepared in step (2), oats are immersed in it, taken out and sterilized, and shiitake mycelium ( Lentinula edodes (Berk.)) inoculated and cultured, dried and pulverized to prepare a fermented product of Hwangchil loquat oat shiitake mycelium; (4) extracting by adding water to the original target height and then concentrating to prepare an original target height extract; (5) roasting astragalus and then adding water to the pulverized astragalus for extraction and concentration to prepare astragalus extract; (6) roasting bokryeong, adding water to the pulverized bokryeong, extracting it, and then concentrating to prepare a bokryeong extract; (7) extracting by adding water to the pulverized bamboo leaves and then concentrating to prepare a bamboo leaf extract; (8) roasting the baekchul and then adding water to the pulverized baekchul for extraction and concentration to prepare a baekchul extract; (9) Hwangchil loquat oat shiitake mycelium fermented product in step (3) for meat, Wongogogo extract prepared in step (4), Astragalus extract prepared in step (5), step (6) Provides a method for producing a snack for companion animals, comprising adding and mixing the prepared bokryeong extract, the bamboo leaf extract prepared in the step (7), and the baekchul extract prepared in the step (8) do.

In addition, the present invention provides a snack for companion animals prepared by the above method.

The companion animal health snack prepared by the method of the present invention has a high content of beta-glucan, essential amino acids and vitamins for companion animals, and when used in the manufacture of healthy snacks for companion animals using fermented shiitake mycelium, nutrients and preference are secured to ensure companion animals. It is suitable for the taste of animals and has an anti-inflammatory effect, so it can provide a healthy snack for companion animals that is beneficial to health.

1 is a schematic diagram of the manufacturing process of the loquat extract powder of the present invention.
Figure 2 is a schematic diagram of the manufacturing process of the turmeric extract powder of the present invention.
Figure 3 is a schematic diagram of the manufacturing process of hwangchil extract powder of the present invention.
4 is a schematic diagram of the manufacturing process of the original target high extract of the present invention.
5 is a schematic diagram of the manufacturing process of the bamboo leaf extract of the present invention.
6 is a schematic diagram of the manufacturing process of the baekchul extract of the present invention.
7 is a schematic diagram of the manufacturing process of the extract of bokryeong of the present invention.
8 is a schematic diagram of the manufacturing process of the astragalus extract of the present invention.
9 is a schematic view of the manufacturing process of the fermented loquat oat shiitake mycelium of the present invention.
Figure 10 is a schematic view of the manufacturing process of the fermented product of hwangchil oat shiitake mycelium of the present invention.
11 is a schematic view of the manufacturing process of the fermented product of Hwangchil loquat oat shiitake mycelium of the present invention.
12 is a schematic view of the manufacturing process of the fermented turmeric oat shiitake mycelium of the present invention.
13 is a graph comparing the cytotoxicity according to the concentration of herbal extracts.
14 is a graph comparing cytotoxicity according to the concentration of fermented oriental medicinal shiitake mycelium.
15 is a graph comparing NO production according to the concentration of fermented shiitake mycelium and healthy snack.
16 is a graph comparing the production rate of GM-CSF according to the concentration of shiitake mycelium fermented product and healthy snack.
17 is a graph comparing the IL-1β production rate according to the concentration of fermented shiitake mycelium and healthy snack.
18 is a schematic diagram of the manufacturing process of a dog health snack.
19 is a schematic diagram of the manufacturing process of a cat healthy snack.

In order to achieve the object of the present invention, the present invention

(1) Roasting loquat, adding water to the pulverized loquat for extraction, concentration and freeze-drying to prepare loquat extract powder;

(2) extracting by adding water to the pulverized hwangchil after roasting hwangchil, and concentrating and freeze-drying to prepare hwangchil extract powder;

(3) After adding water to a mixture of the loquat extract powder prepared in step (1) and the hwangchil extract powder prepared in step (2), oats are immersed in it, taken out and sterilized, and shiitake mycelium ( Lentinula edodes (Berk.)) inoculated and cultured, dried and pulverized to prepare a fermented product of Hwangchil loquat oat shiitake mycelium;

(4) extracting by adding water to the original target height and then concentrating to prepare an original target height extract;

(5) roasting astragalus and then adding water to the pulverized astragalus for extraction and concentration to prepare astragalus extract;

(6) roasting bokryeong, adding water to the pulverized bokryeong, extracting it, and then concentrating to prepare a bokryeong extract;

(7) extracting by adding water to the pulverized bamboo leaves and then concentrating to prepare a bamboo leaf extract;

(8) roasting the baekchul and then adding water to the pulverized baekchul for extraction and concentration to prepare a baekchul extract; and

(9) Hwangchil loquat oat shiitake mycelium fermented product in step (3) for meat, Wongogogo extract prepared in step (4), Astragalus extract prepared in step (5), step (6) Provides a method for producing a snack for companion animals, comprising adding and mixing the prepared bokryeong extract, the bamboo leaf extract prepared in the step (7), and the baekchul extract prepared in the step (8) do.

In the method of manufacturing a snack for companion animals of the present invention, the shiitake mycelium ( Lentinula edodes (Berk.)) is the shiitake mycelium ( Lentinula edodes (Berk.)) JMIF-001 (KCTC18874P), and the strain was applied to the Korea Research Institute of Bioscience and Biotechnology. Deposited on November 27, 2020. The deposited specific shiitake mycelium is a mycelium with high fermentation efficiency selected from the herbal medicine extract powder.

In addition, in the method of manufacturing a snack for companion animals of the present invention, the companion animal may be a dog or a cat, but is not limited thereto.

In addition, in the method of manufacturing a snack for companion animals of the present invention, the meat of step (9) may include meat such as duck meat, pork, beef, chicken, lamb, horse meat, or fish such as tuna, not limited

In addition, in the method of manufacturing a snack for companion animals of the present invention, the extract powder of loquat in step (1) is preferably obtained by roasting loquat at 90 to 110° C. for 2 to 4 minutes and then adding water to the pulverized loquat to obtain 70 It can be prepared by extraction at ~90°C for 8-12 hours, concentration and freeze-drying, and more preferably, roasting loquats at 100°C for 3 minutes, adding water to the pulverized loquats, and extracting at 80°C for 10 hours. And it can be prepared by concentration and freeze-drying.

In addition, in the method for producing a snack for companion animals of the present invention, the extract powder of hwangchil in step (2) is preferably obtained by roasting hwangchil at 140-160° C. for 8-12 minutes and then adding water to the crushed hwangchil 80 It can be prepared by extraction at ~90 ℃ for 7-9 hours, concentration and freeze-drying, more preferably, roasting hwangchil at 150℃ for 10 minutes, then adding water to the crushed hwangchil and extracting at 85℃ for 8 hours. And it can be prepared by concentration and freeze-drying.

In addition, in the method for producing a snack for companion animals of the present invention, the fermented product of hwangchil loquat oat shiitake mycelium in step (3) is preferably a mixture of 12 to 18 g of loquat extract powder and 12 to 18 g of hwangchil extract powder. After adding 1.4~1.6 L of water, oats are immersed here for 50~70 minutes, taken out and sterilized, and inoculated with shiitake mycelium ( Lentinula edodes (Berk.)) JMIF-001(KCTC18874P) at 22~26℃ It can be prepared by culturing for 3 to 5 weeks in the , and then drying and pulverizing, more preferably, after adding 1.47 L of water to a mixture of 15 g of loquat extract powder and 15 g of hwangchil extract powder, and then adding oats to it. After being immersed for 60 minutes, taken out and sterilized, inoculated with shiitake mycelium ( Lentinula edodes (Berk.)) JMIF-001 (KCTC18874P), cultured at 24° C. for 4 weeks, and dried and pulverized. It was possible to prepare a fermented product under the conditions as described above, which has excellent mycelial growth, high beta-glucan, polyphenol and amino acid content, and a more efficient calcium intake.

In addition, in the method for producing a snack for companion animals of the present invention, the original target height extract of step (4) is preferably 7-9 at 70-80°C by adding 80-120 L of water to 8-12 kg of the original target height. After extraction for a period of time, it may be prepared by concentration, and more preferably, 100 L of water is added to 10 kg of the original target, extracted at 75° C. for 8 hours, and then concentrated.

In addition, in the method of manufacturing a snack for companion animals of the present invention, the astragalus extract of step (5) is preferably 4 to 6 kg of astragalus roasted at 140 to 160° C. for 3 to 10 minutes and then pulverized astragalus with water 45 It can be prepared by adding ~55 L, extracting at 70~90℃ for 10~14 hours, and then concentrating, more preferably, roasting 5 kg of astragalus at 150℃ for 3~10 minutes, and then adding 50 L of water to the pulverized astragalus. It can be prepared by adding and extracting at 80° C. for 12 hours and then concentrating.

In addition, in the method for manufacturing a snack for companion animals of the present invention, the extract of bokryeong in step (6) is preferably 4 to 6 kg of bokryeong roasted at 110 to 130° C. for 1-2 minutes and then crushed bokryeong with water 30 It can be prepared by adding ~40 L and extracting it at 90~100℃ for 10~14 hours and then concentrating it. More preferably, 5 kg of bokryeong is roasted at 120℃ for 1 minute and then 35L of water is added to the pulverized bokryeong. It can be prepared by extracting at 95° C. for 12 hours and then concentrating.

In addition, in the method for producing a snack for companion animals of the present invention, the bamboo leaf extract in step (7) is preferably obtained by adding 40-60 L of water to 4-6 kg of crushed bamboo leaves at 70-90°C for 10-14 It can be prepared by time extraction, followed by filtration and concentration, and more preferably, 50 L of water is added to 5 kg of pulverized bamboo leaves, followed by extraction at 80° C. for 12 hours, followed by filtration and concentration.

In addition, in the method of manufacturing a snack for companion animals of the present invention, the baekchul extract in step (8) is preferably roasted for 2-4 minutes at 140-160 ° C. of 4-6 kg of baekchul and then crushed baekchul with water It can be prepared by adding ~45 L and extracting at 80~100℃ for 8~12 hours and then concentrating, more preferably, roasting 5 kg of baekchul at 150℃ for 3 minutes and then adding 40L of water to the pulverized baekchul It can be prepared by extracting at 90° C. for 10 hours and then concentrating.

The manufacturing method of the snack for companion animals of the present invention, more specifically,

(1) Roasting loquats at 90-110° C. for 2-4 minutes, adding water to the pulverized loquats, extracting them at 70-90° C. for 8-12 hours, concentrating and freeze-drying to prepare loquat extract powder;

(2) roasting hwangchil at 140-160° C. for 8-12 minutes, adding water to the pulverized hwangchil, extracting it at 80-90° C. for 7-9 hours, and concentrating and freeze-drying to prepare hwangchil extract powder;

(3) After adding 1.4~1.6 L of water to a mixture of 12~18 g of loquat extract powder prepared in step (1) and 12~18 g of hwangchil extract powder prepared in step (2), here After immersing oats for 50~70 minutes, take them out and sterilize them, inoculate with shiitake mycelium ( Lentinula edodes (Berk.)) JMIF-001(KCTC18874P) and incubate at 22~26℃ for 3~5 weeks, then dry and pulverize to prepare a fermented product of hwangchil loquat oat shiitake mycelium;

(4) adding 80~120 L of water to 8~12 kg of original target height, extracting at 70~80℃ for 7~9 hours, and then concentrating to prepare an original target high extract;

(5) Roast 4-6 kg of astragalus at 140-160℃ for 3-10 minutes, add 45-55 L of water to the pulverized astragalus, extract at 70-90℃ for 10-14 hours, and then concentrate to prepare astragalus extract to do;

(6) After roasting 4~6 kg of bokryeong at 110~130℃ for 1~2 minutes, adding 30~40L of water to the pulverized bokryeong, extracting it at 90~100℃ for 10~14 hours, and concentrating to prepare bokryeong extract to do;

(7) adding 40-60 L of water to 4-6 kg of crushed bamboo leaves, extracting at 70-90° C. for 10-14 hours, and concentrating to prepare a bamboo leaf extract;

(8) After roasting 4-6 kg of baekchul at 140-160℃ for 2-4 minutes, adding 35-45 L of water to the pulverized baekchul, extracting at 80-100℃ for 8-12 hours, and concentrating to prepare a baekchul extract to do; and

(9) Hwangchil loquat oat shiitake mycelium fermented product in step (3) for meat, Wongogogo extract prepared in step (4), Astragalus extract prepared in step (5), step (6) It may include adding and mixing the prepared bokryeong extract, the bamboo leaf extract prepared in the step (7), and the baekchul extract prepared in the step (8),

more specifically

(1) Roasting loquats at 100° C. for 3 minutes, adding water to the pulverized loquats, extracting them at 80° C. for 10 hours, concentrating and freeze-drying to prepare a loquat extract powder;

(2) roasting hwangchil at 150° C. for 10 minutes, adding water to the pulverized hwangchil, extracting at 85° C. for 8 hours, and concentrating and freeze-drying to prepare hwangchil extract powder;

(3) After adding 1.47 L of water to a mixture of 15 g of loquat extract powder prepared in step (1) and 15 g of hwangchil extract powder prepared in step (2), add oats to it for 60 minutes After immersion, take out and sterilize, inoculated with shiitake mycelium ( Lentinula edodes (Berk.)) JMIF-001 (KCTC18874P) and cultured at 24° C. for 4 weeks, dried and pulverized to prepare a fermented product of Hwangchil loquat oat shiitake mycelium ;

(4) adding 100 L of water to 10 kg of original target height, extracting at 75° C. for 8 hours, and then concentrating to prepare an original target high extract;

(5) roasting 5 kg of astragalus at 150° C. for 3 to 10 minutes, adding 50 L of water to the pulverized astragalus, extracting at 80° C. for 12 hours, and then concentrating to prepare an astragalus extract;

(6) roasting 5 kg of bokryeong at 120°C for 1 minute, adding 35 L of water to the pulverized bokryeong, extracting at 95°C for 12 hours, and concentrating to prepare a bokryeong extract;

(7) adding 50 L of water to 5 kg of pulverized bamboo leaves, extracting at 80° C. for 12 hours, and concentrating to prepare a bamboo leaf extract;

(8) roasting 5 kg of baekchul at 150° C. for 3 minutes, adding 40 L of water to the pulverized baekchul, extracting at 90° C. for 10 hours, and concentrating to prepare a baekchul extract; and

(9) Hwangchil loquat oat shiitake mycelium fermented product in step (3) for meat, Wongogogo extract prepared in step (4), Astragalus extract prepared in step (5), step (6) It may include adding and mixing the prepared bokryeong extract, the bamboo leaf extract prepared in the step (7), and the baekchul extract prepared in the step (8).

In the method for producing a snack for companion animals of the present invention, in the case of a dog snack when mixing in step (9), 30-40 g of fermented Hwangchil loquat oat shiitake mycelium, preferably 45-55 g of duck meat, 4-6 g of extract, 1.5-2.5 g of Astragalus extract, 2.5-3.5 g of baekbokryeong extract, 2.5-3.5 g of bamboo leaf extract, and 1.5-2.5 g of baekchul extract can be mixed, and more preferably, 50 g of duck meat with Hwangchil loquat. 35 g of fermented oat shiitake mycelium, 5 g of Wongogogo extract, 2 g of Astragalus extract, 3 g of baekbokryeong extract, 3 g of bamboo leaf extract and 2 g of baekchul extract can be mixed.

In addition, in the case of a cat snack when mixing in step (9) above, 55-65 g of tuna, 20-30 g of fermented Hwangchil loquat oat shiitake mycelium, 2.5-3.5 g of Wongogogo extract, 1.5-2.5 g of Astragalus extract, Baekbokryeong 2.5-3.5 g of extract, 1.5-2.5 g of bamboo leaf extract, and 4.5-5.5 g of baekchul extract can be mixed, and more preferably, 25 g of fermented Hwangchil loquat oat shiitake mycelium in 60 g of tuna, 3 g of Wongogogo extract, and Hwanggi 2 g of extract, 3 g of baekbokryeong extract, 2 g of bamboo leaf extract and 5 g of baekchul extract may be mixed.

Mixing under the same conditions as described above is preferred by companion animals, and has a high beta-glucan, polyphenol, amino acid and vitamin content and anti-inflammatory effect, making it a more beneficial snack for companion animals.

The present invention also provides a snack for companion animals prepared by the above method.

Hereinafter, the present invention will be described in detail by way of preparation examples and examples. However, the following Preparation Examples and Examples only illustrate the present invention, the content of the present invention is not limited to the following Preparation Examples and Examples.

Preparation Example 1. Preparation of extract powder of loquat, turmeric and hwangchil

(1) The manufacturing process of the loquat extract powder is as shown in FIG. Loquat was roasted at 100° C. for 3 minutes and then pulverized to 100 mesh using a blender. Purified water was added 10 times (v/w) to the pulverized loquat, extracted at 80° C. for 10 hours, filtered using a nonwoven fabric, and then cooled at room temperature. The loquat extract cooled to room temperature was concentrated to 2 brix and lyophilized to prepare a loquat extract powder.

(2) The manufacturing process of turmeric extract powder is as shown in FIG. After roasting turmeric at 120° C. for 5 minutes, it was pulverized to 120 mesh using a blender. Purified water was added 10 times (v/w) to the pulverized turmeric, extracted at 90° C. for 8 hours, filtered using a nonwoven fabric, and then cooled at room temperature. The turmeric extract cooled to room temperature was concentrated to 1 brix and freeze-dried to prepare a turmeric extract powder.

(3) The manufacturing process of hwangchil extract powder is as shown in FIG. After roasting hwangchil at 150° C. for 10 minutes, it was pulverized to 120 mesh using a blender. Purified water was added 10 times (v/w) to the pulverized hwangchil, extracted at 85° C. for 8 hours, filtered using a nonwoven fabric, and then cooled at room temperature. Hwangchil extract cooled to room temperature was concentrated to 2 brix and freeze-dried to prepare hwangchil extract powder.

Preparation Example 2. Preparation of Wongoal Goji, Bamboo Leaf, Baekchul, Bokryeong, Astragalus Extract

(1) The manufacturing process of the original target height extract is as shown in FIG. 4 . After grinding the original target height of 10 kg to 100 mesh using a blender, 100 L of purified water was added and extracted at 75° C. for 8 hours. The extracted extract was filtered using a nonwoven fabric, and then concentrated to 10 brix to prepare an original target high extract.

(2) The manufacturing process of the bamboo leaf extract is as shown in FIG. 5 . After grinding 5 kg of bamboo leaves to 80 mesh using a blender, 50 L of purified water was added and extracted at 80° C. for 12 hours. The extracted extract was filtered using a nonwoven fabric and concentrated to 5 brix to prepare a bamboo leaf extract.

(3) The manufacturing process of the baekchul extract is as shown in FIG. After roasting 5 kg of baekchul at 150° C. for 3 minutes, it was pulverized to 100 mesh using a blender. 40 L of purified water was added to the pulverized baekchul, extracted at 90° C. for 10 hours, filtered using a nonwoven fabric, and concentrated to 5 brix to prepare a baekchul extract.

(4) The manufacturing process of bokryeong extract is as shown in FIG. After roasting 5 kg of bokryeong at 120° C. for 1 minute, it was pulverized to 120 mesh using a blender. 35 L of purified water was added to the pulverized bokryeong, extracted at 95° C. for 12 hours, filtered using a nonwoven fabric, and concentrated to 5 brix to prepare a bokryeong extract.

(5) The manufacturing process of Astragalus extract is as shown in FIG. After roasting 5 kg of astragalus at 150° C. for 3 to 10 minutes, it was ground to 80 mesh using a blender. 50 L of purified water was added to the pulverized astragalus, extracted at 80° C. for 12 hours, filtered using a nonwoven fabric, and concentrated to 5 brix to prepare an astragalus extract.

Preparation Example 3. Preparation of fermented loquat oat shiitake mycelium, hwangchil oat shiitake mycelium fermented product, hwangchil loquat oat shiitake mycelium fermented product, turmeric oat shiitake mycelium fermented product

(1) 30 g of loquat extract powder (step (1) of Preparation Example 1) was dissolved in 1.47 L of water, 1 kg of dried oats was added thereto, immersed for one hour, and then taken out. The oats were sterilized in an autoclave at 121° C. for 35 minutes in an autoclave at 121° C. to remove contaminants after dividing the oats in a certain amount in mycelial culture bags. After cooling the sterilized oats on a clean bench, the shiitake mycelium (KCTC18874P) grown in PDA (Potato dextrose agar, DifcoTM, USA) medium was aseptically inoculated and inoculated in an incubator at 24° C. for 4 weeks. After culturing was completed, sterilization was performed for 15 minutes in an autoclave at 80° C. for safety of raw materials, the vinyl was removed, and drying was performed in a dry oven at 70° C. Then, the obtained dried product was pulverized to prepare a fermented product of loquat oat shiitake mycelium (FIG. 9).

(2) 30 g of hwangchil extract powder (step (3) of Preparation Example 1) was dissolved in 1.47 L of water, 1 kg of dried oats was added thereto, immersed for one hour, and then taken out. The oats were sterilized in an autoclave at 121° C. for 35 minutes in an autoclave at 121° C. to remove contaminants after dividing the oats in a certain amount in mycelial culture bags. After cooling the sterilized oats on a clean bench, the shiitake mycelium (KCTC18874P) grown in PDA (Potato dextrose agar, DifcoTM, USA) medium was aseptically inoculated and inoculated in an incubator at 24° C. for 4 weeks. After culturing was completed, sterilization was performed for 15 minutes in an autoclave at 80° C. for safety of raw materials, the vinyl was removed, and drying was performed in a dry oven at 70° C. Then, the obtained dried product was pulverized to prepare a fermented product of hwangchil oat shiitake mycelium (FIG. 10).

(3) Dissolve in 1.47 L of water in a mixture of 15 g of loquat extract powder ((1) step of Preparation Example 1) and 15 g of hwangchil extract powder ((3) step of Preparation Example 1), and dried oats here 1 kg was added, immersed for one hour, and then taken out. The oats were sterilized in an autoclave at 121° C. for 35 minutes in an autoclave at 121° C. to remove contaminants after dividing the oats in a certain amount in mycelial culture bags. After cooling the sterilized oats on a clean bench, the shiitake mycelium (KCTC18874P) grown in PDA (Potato dextrose agar, DifcoTM, USA) medium was aseptically inoculated and inoculated in an incubator at 24° C. for 4 weeks. After culturing was completed, sterilization was performed for 15 minutes in an autoclave at 80° C. for safety of raw materials, the vinyl was removed, and drying was performed in a dry oven at 70° C. Then, the obtained dried product was pulverized to prepare a fermented product of hwangchil loquat oat shiitake mycelium (FIG. 11).

(4) 1.485 L of water was dissolved in 15 g of turmeric extract powder (step (2) of Preparation Example 1), 1 kg of dried oats was added thereto, immersed for one hour, and then taken out. The oats were sterilized in an autoclave at 121° C. for 35 minutes in an autoclave at 121° C. to remove contaminants after dividing the oats in a certain amount in mycelial culture bags. After cooling the sterilized oats on a clean bench, the shiitake mycelium (KCTC18874P) grown in PDA (Potato dextrose agar, DifcoTM, USA) medium was aseptically inoculated and inoculated in an incubator at 24° C. for 4 weeks. After culturing was completed, sterilization was performed for 15 minutes in an autoclave at 80° C. for safety of raw materials, the vinyl was removed, and drying was performed in a dry oven at 70° C. Then, the obtained dried product was pulverized to prepare a fermented product of turmeric and oat shiitake mycelium (FIG. 12).

Preparation Example 4. Dog and cat healthy snack snack manufacturing

(1) Dog snack development

For dog snack development, duck tenderloin, Hwangchil loquat oat shiitake mycelium fermented product (Step (3) of Preparation Example 3), Wongogogo extract (Step (1) of Preparation Example 2), Astragalus extract ((5) of Preparation Example 2) step), bokryeong extract (step (4) of Preparation Example 2), bamboo leaf extract (step (2) of Preparation Example 2), and baekchul extract (step (3) of Preparation Example 2) by varying the mixing ratio of dog 1, dog 2, dog 3, dog 4, and dog 5 were produced and sensory evaluation was performed. As a result, duck tenderloin 50 g, Hwangchil loquat oat shiitake mycelium fermented product 35 g, Wongogogo extract 5 g, Astragalus extract 2 g, Dog 3, which was prepared with a mixing ratio of 3 g of baekbokryeong extract, 3 g of bamboo leaf extract, and 2 g of baekchul extract, showed the highest preference, and was selected as the final mixing ratio.

The dog treat development process involves adding 35 g of fermented Hwangchil loquat oat shiitake mycelium to 50 g of ground duck tenderloin, 5 g of Wongogogo extract, 2 g of Astragalus extract, 3 g of Baekbokryeong extract, 3 g of bamboo leaf extract, and 2 g of baekchul extract. Blended for 10 minutes with a stirrer. The formulated contents were injected into a retort pouch, sterilized at 123° C. for 30 minutes, and then allowed to cool at room temperature to prepare dog health snacks (Tables 1 and 2).

Dog health snack prototype mixing ratio (wt%) division dog 1 dog 2 dog 3 dog 4 dog 5 duck tenderloin 30 40 50 60 70 Hwangchil Loquat Oat Shiitake Mycelium Fermented Product 55 45 35 25 15 original target height extract 5 5 5 5 5 Astragalus extract 2 2 2 2 2 bokryeong extract 3 3 3 3 3 bamboo leaf extract 3 3 3 3 3 baekchul extract 2 2 2 2 2 Sum 100 100 100 100 100

Dog health snack prototype sensory test division intake full symbol Comparative example (commercial product) 7.1 7.2 dog 1 6.9 6.8 dog 2 7.3 7.4 dog 3 8.1 8.1 dog 4 7.6 7.5 dog 5 7.1 7.1

Very good 9 points, good 7 points, average 5 points, bad 3 points, very bad 1 point

(2) Cat snack development

For cat snack development, fermented tuna, hwangchil loquat oat shiitake mycelium (step (3) of Preparation Example 3), Wongogogo extract (Step (1) of Preparation Example 2), Astragalus extract (Step (5) of Preparation Example 2) ), bokryeong extract (step (4) of Preparation Example 2), bamboo leaf extract (step (2) of Preparation Example 2), and baekchul extract (step (3) of Preparation Example 2) by varying the mixing ratio of Cat 1 and Cat 2 , Cat 3, Cat 4 and Cat 5 prototypes were produced and sensory evaluation was performed. As a result, 60 g of tuna, 25 g of fermented Hwangchil loquat oat shiitake mycelium, 3g of Wongogogo extract, 2 g of Astragalus extract, 3 g of Baekbokryeong extract , 2 g of bamboo leaf extract, and 5 g of baekchul extract showed the highest preference for Cat 4, which was selected as the final mixing ratio.

In the cat snack development process, 60 g of crushed tuna, 25 g of fermented Hwangchil loquat oat shiitake mycelium, 3 g of Wongogogo extract, 2 g of Astragalus extract, 3 g of Baekbokryeong extract, 2 g of bamboo leaf extract, and 5 g of baekchul extract were added with a stirrer. Blended for 10 minutes. The formulated contents were injected into a retort pouch, sterilized at 123° C. for 30 minutes, and then allowed to cool at room temperature to prepare healthy cat snacks (Tables 3 and 4).

Cat health snack prototype mixing ratio (wt%) division cat 1 cat 2 cat 3 cat 4 cat 5 tuna 30 40 50 60 70 Hwangchil Loquat Oat Shiitake Mycelium Fermented Product 55 45 35 25 15 original target height extract 3 3 3 3 3 Astragalus extract 2 2 2 2 2 bokryeong extract 3 3 3 3 3 bamboo leaf extract 2 2 2 2 2 baekchul extract 5 5 5 5 5 Sum 100 100 100 100 100

Cat health snack prototype sensory test division intake full symbol Comparative example (commercial product) 6.6 6.7 cat 1 6.4 6.5 cat 2 6.8 6.8 cat 3 7.1 7.2 cat 4 7.9 7.8 cat 5 7.0 7.1

Very good 9 points, good 7 points, average 5 points, bad 3 points, very bad 1 point

2. Experimental method

(1) Selection of optimal fermented shiitake mycelium

Table 5 shows the results of measuring the growth of shiitake mycelium by measuring the length of the mycelium grown downward from the inoculation surface in order to measure the fermentation efficiency for each sample and each mushroom mycelium. JMI-10075 mycelium was not fermented in all herbal medicine extract powder addition groups including the control group. JMI-10071 and JMI-10079 hyphae were fermented at a concentration of 10 to 200 g/L of each sample, but the length of the hyphae was 2-3 cm, indicating weak growth. The JMI-10076 mycelium showed a mycelial length of 4 to 6 cm at the concentrations of relatively 10 g/L, 20 g/L, and 50 g/L, indicating good fermentation. KCTC 18874P mycelium showed excellent mycelial fermentation in all samples except for the high concentration of 300 g/L. Therefore, KCTC 18874P strain ( Lentinula edodes (Berk.) JMIF-001) was selected and used for the preparation of fermented shiitake mycelium extracts from herbal medicines.

(2) Selection of optimal shiitake mycelium fermentation temperature

The results of measuring the optimal fermentation temperature of Shiitake hyphae KCTC 18874P are shown in Table 6 below. As a result of temperature treatment from 16°C to 32°C at intervals of 2°C, mycelial fermentation was good at 22°C and 25°C, and mycelial fermentation was excellent at 24°C. Accordingly, the mycelial fermentation temperature was set to 24° C. when preparing fermented shiitake mycelium extracts from herbal medicines.

Shiitake mycelium (KCTC 18874P) optimum fermentation temperature Temperature (℃) Mycelial growth (25 days) 16 - ^a 18 - 20 + 22 ++ 24 +++ 26 ++ 28 + 30 + 32 -

^a Mycelial fermentation (-: no mycelial fermentation, +: mycelial fermentation weak, ++: mycelial fermentation good, +++: mycelial fermentation excellent). Mycelium no fermentation: less than 1 cm, mycelial fermentation weak: 2-3 cm, mycelial fermentation good: 4-6 cm, mycelial fermentation excellent: 7 cm or more

(3) Beta glucan analysis

- Grain Beta Glucan

Accurately weigh 80-120 mg of the crushed sample, put it in a tube, add 0.2 mL of 50% ethanol and 4 mL of 20 mM sodium phosphate buffer, vortex, and immediately incubate for 3 minutes in a water bath at 100°C. Then, after incubation for 5 minutes in a 50 ℃ constant temperature water bath, 0.2 mL of lichenase was added and mixed, the cap was sealed, and incubated in a 50 ℃ constant temperature water bath for 1 hour. Then, 5 mL of 200 mM sodium acetate buffer was added, vortexed, cooled to room temperature, and centrifuged. Immediately take 0.1 mL of the supernatant obtained after centrifugation, add 0.1 mL of β-glucosidase to the reaction tube, add 0.1 mL of 50 mM acetate buffer to the blank reaction tube, incubate at 50°C for 10 minutes, and then 3 mL of GOPOD was added and incubated at 50° C. for 20 minutes, and the absorbance was measured at 510 nm.

- Mushroom Beta Glucan

Beta-glucan of the sample was measured using Megazyme kit (Mushroom and Yeast β-glucan Assay Procedure K-YBGL, Megazyme, Ireland).

First, the total glucan was decomposed by putting 100 mg of a pulverized sample filtered through a 100 mesh sieve into a tube, adding 1.5 mL of 37% HCl, and putting it in a constant temperature water bath at 30° C. for 45 minutes. Then, 10 mL of distilled water was added, vortexed, and incubated at 100° C. for 2 hours. Then, while cooling at room temperature, 10 mL of 2N KOH was added, adjusted to 100 mL with 200 mM sodium acetate buffer, and then thoroughly mixed. After that, exo-1,3-β-glucanase plus β-glucosidase was dissolved in 200 mM sodium acetate buffer in 0.1 mL of the supernatant. Add 0.1 mL of reagent blank, add 0.2 mL of acetate buffer, and for D-glucose standard, add 0.1 mL of D-glucose standard and 0.1 mL of acetate buffer, mix, incubated for minutes. Then, 3 mL of GOPOD (glucose oxidase/peroxidase mixture) was added and incubated at 40° C. for 20 minutes, and absorbance was measured at 510 nm.

For α-glucan (α-Glucan), 100 mg of a pulverized sample sieved through a 100 mesh sieve was put into a tube, 2 mL of 2M KOH was added, and mixed for 20 minutes. After adding 8 mL of 1.2M sodium acetate buffer and mixing, 0.2 mL of amyloglucosidase plus invertase was added, mixed well, and incubated in a constant temperature water bath at 40° C. for 30 minutes. In 0.1 mL of the supernatant, 0.1 mL of 200 mM sodium acetate buffer and 3 mL of GOPOD were added, incubated at 40° C. for 20 minutes, and the absorbance was measured at 510 nm. The calculation quantified the content of β-glucan by subtracting the α-glucan content from the total glucan content.

(4) Determination of total polyphenol content

Analysis of total polyphenol content was tested by Folin-Denis method. After diluting the sample 1000 times, 0.025 mL of Folin reagent was added, and after 6 minutes, 0.1 mL of a saturated Na ₂ CO ₃ solution was added and mixed to develop color. After 1 hour, the absorbance of the colored reagent was measured at 765 nm, and the standard material was treated with gallic acid at 100, 200, 300, 400 and 500 μg/mL, and then a standard curve showing the relationship between absorbance and concentration was prepared. Total polyphenol content is shown.

(5) amino acids

For amino acid analysis, put 0.5 g of the sample into a test tube, add 10 mL of 6 N-HCl, melt the tip of the test tube with fire to make an ampoule, seal it, and then hydrolyze it in an autoclave at 110° C. for 24 hours. mL, concentrated under reduced pressure, and adjusted with 5 mL of 20 mM HCl. A certain amount of the filtrate obtained by filtration through a 0.45 μm membrane filter was derivatized using AccQ-Tag reagent and analyzed by HPLC. The analysis conditions are shown in Table 7.

Amino Acid Analysis Conditions Item Analysis conditions equipment Agilent Technologies 1200 Series detector Agilent Technologies 1200 Series FLDs column AccQ-Tag™
(Waters Co., 150 mm L × 3.9 mm ID) column temperature 37℃ buffer solution A: AccQ-Tag Eluent A (acetate-phosphate buffer)
B: AccQ-Tag Eluent B (100% acetonitrile)
C: DW hour A B C 0 100 0 0 0.5 99 One 0 18 95 5 0 19 91 9 0 26 86.7 13.3 0 30 84 16 0 32 83 17 0 36 0 60 40 39 100 0 0 48 100 0 0 flow rate 1.0 mL/min infusion dose 5 μl

(6) Vitamin B1, B2

Weigh 0.5 g of the sample accurately, add 2 mL of distilled water, vortex for 10 minutes, add 6 mL of acetonitrile, mix for 20 minutes, and centrifuge at 4000 rpm for 10 minutes. The supernatant was taken and dried under reduced pressure, followed by quantification with 10 mL of distilled water. The total amount of the solution was filtered through a 0.45 μm filter, and 20 μl of it was injected into HPLC (Table 8).

Vitamin B1, B2 Analysis Conditions Item Analysis conditions Instrument Agilent Technologies 1200 Series Column Agilent XDB-C ₁₈ (Method Development Kit)
(4.6 × 150 mm, 5 um) mobile phase A: 20 mM KH ₂ PO ₄ (pH 2.95)
B: 20 mM KH ₂ PO ₄ (pH 2.95)/ACN = 50/50 (v/v) fuel flow 0.8~1.0 mL/min Column temp. 30℃ Wavelength (nm) 275 nm Injection volume 20 μl gradient program Time Flow (mL/min) A (%) B(%) 0.0 0.8 100 0 10.0 0.8 100 0 12.0 1.0 100 0 60.0 1.0 50 50

(7) Calcium analysis

Calcium in the sample is pretreated and analyzed by wet decomposition, and the analysis conditions are shown in Table 9 below. That is, add 10 mL of concentrated nitric acid to 0.5 g of the sample, heat it to a low temperature at first, and decompose it while heating to a high temperature gradually (Buchi distillation Unit B-324). After adjusting to mL, the filtrate was used as a sample for analysis. Quantification of calcium was analyzed by preparing a standard calibration curve by preparing standard solution concentrations of each element at 1, 3, and 5 ppm with an atomic absorption spectrophotometer (Perkin Elmer Analyst 400, USA).

Calcium Analysis Item Analysis conditions Instrument Atomic Absorption Spectrophotometer
(Perkin Elmer Analyst 400) fuel flow C ₂ H ₂ , 2.0/min Oxidant flow Air, 10.0 L/min Wavelength (nm) Ca: 422.67

(8) Analysis of phosphorus (P) content

For the phosphorus component, 1 g of the sample was incinerated at 600°C to obtain white ash, 10 mL of concentrated hydrochloric acid diluted 2 times the ash was added, filtered, evaporated to dryness in a water bath, 10 mL of hydrochloric acid diluted 4 times was added, and distilled water was added. was added and adjusted to 100 mL. To a predetermined amount of the pretreated sample, 1 mL of an ammonium metavanadate solution as a coloring agent was added, and 10 mL of distilled water was used. After standing for 30 minutes, absorbance was measured at 470 nm with a UV spectrophotometer and quantified.

(9) heavy metals

Heavy metals were pretreated and analyzed by dry painting, and the analysis conditions are shown in Table 10. That is, 0.5 g of the sample was incinerated at 600°C to obtain white ash, 10 mL of concentrated hydrochloric acid diluted 2 times was added, filtered, evaporated to dryness in a water bath, 10 mL of hydrochloric acid diluted 4 times was added, and 100% using distilled water. The filtrate, which was aliquoted in mL, was used as an analysis sample. Quantification of each inorganic component was analyzed by preparing a standard calibration curve by preparing standard solution concentrations of each element at 0.5, 1.0, 3.0 and 5.0 mg/kg with an atomic absorbance spectrophotometer (Perkin Elmer AAnalyst 400).

Atomic Absorption Colorimetric Analysis Conditions for Heavy Metal Analysis Content Analysis condition Instrument Atomic Absorption spectrophotometer
(Perkin Elmer AAnalyst 400) fuel flow C ₂ H ₂ , 2.0 mL/min Oxidant flow Air, 10.0 mL/min Wavelength (nm) Pb:283.31, Cd:228.80

(10) cell culture

RAW 264.7 macrophages cell culture was performed using a cell culture medium supplemented with DMEM (Dulbecco Modified Eagle Medium, Welgene, Gyeongsan, Korea) with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin. C incubated in a humidified CO ₂ incubator (5% CO ₂ /95% air). When the cells were about 80% full of the culture dish, the monolayer of the cells was washed with PBS (phosphate-buffered saline, pH 7.4), subcultured with 0.25% trypsin-2.65 mM EDTA, and the medium was changed every 2 days.

(11) Confirmation of cell viability

Cytotoxicity was measured by MTT assay. Each cell was cultured in a 96-well plate at a concentration of 1 × 10 ⁵ cell/mL and used in the experiment. After each sample was treated at a concentration of 100, 500, and 1000 μg/mL for 24 hours, the medium was removed, and 10 μl MTS was added to the new medium, reacted for 4 hours, and absorbance was measured at 540 nm. Cell viability of each sample was calculated relative to the sample untreated group as 100%.

(12) Measurement of NO (nitric oxide) production

Raw 264.7 cells were aliquoted to a 96-well plate at 1×10 ⁵ cell/mL per well and cultured for 24 hours, followed by treatment with the sample and LPS (lipopolysaccharide). LPS was treated at a concentration of 1 μg/mL, respectively, and each sample was diluted to a concentration of 100, 500, and 1000 μg/mL and added to the cells. After 24 hours, the cell culture medium was recovered and the NO production inhibitory ability was measured using a NO assay using the Griess reagent system. The NO production rate of each sample was calculated relative to the LPS-treated group as 100%.

(13) Measurement of inflammatory cytokines (GM-CSF, IL-1β) production

The amount of inflammatory cytokine production in the cell culture was measured using a mouse enzyme-linked immnunosorbent assay (ELISA) kit. Raw 264.7 cells were adjusted to 5×10 ⁵ cells/ml using DMEM medium, and then inoculated into a 6-well plate, and cultured for 24 hours in a 5% CO ₂ incubator. Cells were treated with 1 μg/mL of LPS, and after 1 hour, each sample was treated by concentration and cultured for 24 hours. The pro-inflammatory cytokine content of the supernatant obtained after incubation was measured, and quantification was quantified using an ELISA kit, and the standard curve R ² value for the standard was 0.99 or more.

(14) Sensory evaluation

In the sensory test, the amount of intake consumed by the companion animal was compared, the higher the intake, the higher the score, and the overall preference was scored by comparing the number of times the companion animal chose among the five snacks and the degree of preference. Scoring criteria were very good 9 points, good 7 points, average 5 points, bad 3 points, and very bad 1 point.

(15) Statistical processing

The average value and standard deviation between each experimental group were calculated using the SPSS statistical analysis program (SPSS, ver. 16.0, USA) for the results of this experiment, and the statistical significance was p <0.05. was verified with

Example 1. Beta Glucan

The beta-glucan content of each herbal medicine fermented shiitake mycelium is shown in Table 11. (1-3)(1-4) β-glucan is mainly found in cereals, and (1-3)(1-6) β-glucan is detected in yeast and mushrooms. Only (1-3)(1-6) β-glucan is known to have anticancer and immune enhancing actions. (1-3)(1-4) β-glucan showed the content of 1.07~1.81%, and the content of turmeric oat shiitake mycelium fermented product was 1.81%, showing the highest content. (1-3)(1-6) β-glucan showed the content of 43.25~48.58%, and the content of Hwangchil loquat oat shiitake mycelium fermented product was 48.58%, showing the highest content.

Fermented product beta-glucan content (%) sample (1-3)(1-4) β-glucan (1-3)(1-6) β-glucan Total Beta Glucans Loquat oat shiitake mycelium fermented product 1.07 43.25 44.32 Hwangchil oat shiitake mycelium fermented product 1.16 43.64 44.8 Hwangchil Loquat Oat Shiitake Mycelium Fermented Product 1.55 48.58 50.13 Fermented product of turmeric oat shiitake mycelium 1.81 47.08 48.89

Table 12 shows the contents of (1-3)(1-4) β-glucan and (1-3)(1-6) β-glucan of the dog prototype to which the fermented herbal medicine extract mycelium was added. The contents of (1-3)(1-4) β-glucan and (1-3)(1-6) β-glucan in the existing marketed dog treat products were 0.16 and 8.34%, respectively, and the total beta-glucan content was 8.5%. it was The contents of (1-3)(1-4) β-glucan and (1-3)(1-6) β-glucan in the dog prototype were 0.97 to 1.36% and 11.74 to 24.72%, respectively, which were significantly different from the existing commercial products. showed an increase in (1-3)(1-4) β-glucan content is 1.36%, 50 g of duck tenderloin, 35 g of fermented Hwangchil loquat oat shiitake mycelium, 5 g of Wongogogo extract, 2 g of Astragalus extract, 3 g of Baekbokryeong extract, bamboo leaf Dog 3 prepared with a mixing ratio of 3 g of extract and 2 g of baekchul extract showed the highest level, and (1-3)(1-6) β-glucan content also showed the highest level in Dog 3. Therefore, dog 3 was selected as the final dog prototype.

Dog health snack beta-glucan content (%) sample (1-3)(1-4) β-glucan (1-3)(1-6) β-glucan Total Beta Glucans Comparative example (commercial product) 0.16 8.34 8.5 dog 1 0.97 21.27 22.24 dog 2 1.07 20.53 21.6 dog 3 1.36 24.72 26.08 dog 4 1.05 17.31 18.36 dog 5 1.09 11.74 12.83

Table 13 shows the contents of (1-3)(1-4) β-glucan and (1-3)(1-6) β-glucan in the cat prototype to which the oriental medicine Shiitake mycelium fermented product was added. The content of (1-3)(1-4) β-glucan and (1-3)(1-6) β-glucan in the previously marketed cat snack products was 0.71 and 8.51%, respectively, and the total beta-glucan content was 9.22 was %. The contents of (1-3)(1-4) β-glucan and (1-3)(1-6) β-glucan of the cat prototype were 1.25-2.12 and 15.24-21.98%, respectively, which were significantly different than those of the existing commercial products. showed an increase. (1-3)(1-4) β-glucan content is 2.12%, 60 g of tuna, 25 g of fermented turmeric oat shiitake mycelium, 3 g of Wongogo extract, 2 g of Astragalus extract, 3 g of Baekbokryeong extract, 2 bamboo leaf extract g and 5 g of baekchul extract showed the highest level in Cat 4, and the (1-3)(1-6) β-glucan content was 21.98%, which was the highest in Cat 4. Therefore, cat 4 was selected as the final cat prototype.

Cat health snack beta-glucan content (%) sample (1-3) (1-4) β-glucan (1-3) (1-6) β-glucan Total Beta Glucans Comparative example (commercial product) 0.71 8.51 9.22 cat 1 2.01 15.24 17.25 cat 2 1.89 16.11 18 cat 3 1.64 18.41 20.05 cat 4 2.12 21.98 24.1 cat 5 1.25 17.86 19.11

Example 2. Total polyphenol content

The total polyphenol content of oat fermented products containing loquat, hwangchil, and turmeric extracts fermented with shiitake mycelium was high in turmeric oat shiitake mycelium and Hwangchil loquat oat shiitake mycelium (Table 14).

Fermentation Total Polyphenol Content (mg GAE/g) sample total polyphenols Loquat oat shiitake mycelium fermented product 34.8±2.0 Hwangchil oat shiitake mycelium fermented product 48.5±1.5 Hwangchil Loquat Oat Shiitake Mycelium Fermented Product 52.9±0.6 Fermented product of turmeric oat shiitake mycelium 53.3±2.2

Table 15 shows the total polyphenol content of the dog prototype to which the oriental medicine Shiitake mycelium fermented product was added. The total polyphenol content of previously marketed dog health snack products was found to be 5.8 mg/g, but in dogs 1 to 5 prepared in the present invention, the content was 16.8 to 21.4 mg/g. Dog 3 showed the highest total polyphenol content at 21.4 mg/g.

Dog health snack Total polyphenol content (mg GAE/g) sample total polyphenols Comparative example (commercial product) 5.8±0.9 dog 1 17.8±1.4 dog 2 18.8±1.2 dog 3 21.4±1.1 dog 4 16.8±1.7 dog 5 17.2±1.4

Table 16 below shows the total polyphenol content of the cat prototype to which the fermented oriental herbal extract mycelium was added. As a result of this study, it was confirmed that the total polyphenol content of the prepared cat healthy snacks was 21.7~31.2 mg/g, which was more than twice higher than the total polyphenol content of 10.1 mg/g of the previously marketed cat snack products. In particular, cat 4 showed a higher level of total polyphenol content than other prototypes, so cat 4 was selected as the final cat prototype.

Cat Healthy Snack Total Polyphenol Content (mg GAE/g) sample total polyphenols Comparative example (commercial product) 10.1±0.7 cat 1 22.4±1.6 cat 2 21.7±1.5 cat 3 26.3±1.2 cat 4 31.2±1.7 cat 5 27.2±1.4

Example 3. Amino Acid Content

The amino acid content of each herbal medicine shiitake mycelium fermented product is shown in Table 17 below. The total amino acid content of each fermented product was in the range of 66.12 ~ 83.79 mg/g, showing differences according to the added herbal medicine. Among them, Hwangchil loquat oat shiitake mycelium fermented product and turmeric oat shiitake mycelium fermented product showed the highest amino acid content in that order.

Fermentation essential amino acid content (mg/g, dry basis) amino acids Loquat oat shiitake mycelium fermented product Hwangchil oat shiitake mycelium fermented product Hwangchil Loquat Oat Shiitake Mycelium Fermented Product Fermented product of turmeric oat shiitake mycelium Arginine 10.21 11.34 14.23 13.48 Histidine 6.21 7.04 9.72 9.07 Isoleucine 8.73 8.42 10.02 9.34 Leucine 14.46 16.82 19.34 18.02 Lysine 6.12 6.34 6.87 6.14 Methionine 3.21 4.02 3.87 3.78 Phenylalanine 7.01 6.87 8.01 7.98 Threonine 3.84 3.41 4.02 3.74 Tryptophan 1.02 1.32 1.47 1.28 Valine 5.31 4.89 6.24 5.73 Sum 66.12 70.47 83.79 78.56

The amino acid content of the dog prototype to which fermented oriental medicinal shiitake mycelium was added is shown in Table 18 below. The amino acid content of existing commercially available dog snack products was 31.01 mg/g, and the dog products manufactured in the present invention showed a content of 65.93 to 74.85 mg/g, indicating a higher content of essential amino acids for dogs than commercial prototypes. . Essential amino acid content is 50 g of duck tenderloin, 35 g of fermented Hwangchil loquat oat shiitake mycelium, 5 g of Wongogogo extract, 2 g of Astragalus extract, 3 g of Baekbokryeong extract, 3 g of bamboo leaf extract, and 2 g of baekchul extract. 3 showed the highest value.

Essential amino acid content for dog health snacks (mg/g, dry basis) Samples comparative example
(commercial product) dog 1 dog 2 dog 3 dog 4 dog 5 Arginine 3.24 6.32 7.23 8.32 8.24 7.01 Histidine 3.44 4.26 4.61 4.26 5.21 6.23 Isoleucine 4.71 5.21 6.32 7.21 7.61 6.46 Leucine 5.26 10.14 9.14 12.14 8.21 9.06 Lysine 1.78 9.34 10.34 11.34 9.36 8.27 Methionine 4.45 4.26 5.26 6.34 6.28 5.97 Phenylalanine 3.46 7.35 6.73 6.35 6.14 5.87 Threonine 2.21 6.71 5.42 5.71 5.41 6.01 Tryptophan 0.34 5.02 5.15 5.84 6.32 6.14 Valine 2.12 7.32 6.93 7.34 7.01 6.78 Sum 31.01 65.93 67.13 74.85 69.79 67.8

The amino acid content of the cat prototype to which the oriental medicinal shiitake mycelium fermented product was added is shown in Table 19 below. The amino acid content of conventionally marketed cat snack products was 37.55 mg/g, and the cat products manufactured in the present invention showed a content of 65.2 to 97.33 mg/g, indicating a higher content of essential amino acids for dogs than commercial prototypes. . The essential amino acid content was found in cat 4 prepared by mixing 60 g of tuna, 25 g of fermented turmeric oat shiitake mycelium, 3 g of Wongogogo extract, 2 g of Astragalus extract, 3 g of Baekbokryeong extract, 2 g of bamboo leaf extract, and 5 g of baekchul extract. It showed the highest level, and the content of taurine, known as an important element among essential amino acids in cats, was also highest in cat 4.

Essential amino acid content of healthy cat snack (mg/g, dry basis) Samples comparative example
(commercial product) cat 1 cat 2 cat 3 cat 4 cat 5 Arginine 4.32 8.34 8.42 10.46 11.43 12.34 Histidine 3.87 5.07 6.02 8.21 9.47 13.01 Isoleucine 3.65 4.62 4.97 7.01 7.98 11.03 Leucine 4.21 12.03 14.25 17.63 17.24 7.21 Lysine 3.98 7.78 11.03 16.31 14.35 11.02 Methionine 3.21 4.21 3.26 2.31 4.15 5.21 Phenylalanine 1.71 6.87 6.43 6.24 7.73 6.21 Threonine 3.23 5.01 6.04 3.01 7.42 5.24 Tryptophan 1.04 1.07 1.36 1.04 2.53 3.21 Valine 6.27 6.42 7.46 7.72 10.21 11.71 Taurine 2.06 3.78 4.17 4.35 4.82 4.26 Sum 37.55 65.2 73.41 84.29 97.33 90.45

Example 4. Vitamin B1, B2 content

The vitamin B1 and B2 contents of each herbal extract mycelium fermented product are shown in Table 20 below. The content of vitamin B1 in the fermented product ranged from 6.16 to 11.21 mg/kg, showing differences according to the herbal medicines. Among them, the fermented turmeric oat shiitake mycelium showed the significantly highest content, followed by Hwangchil loquat oat shiitake mycelium fermented product and loquat oat shiitake mycelium fermented product. The total content of vitamin B2 in the fermented mycelium according to the herbal extracts was in the range of 26.27 to 31.78 mg/kg, and, like the content of B1, the highest content in the fermented turmeric oat shiitake mycelium was shown.

Fermented product vitamin B1, B2 content (mg/kg) sample vitamin B1 vitamin B2 Sum Loquat oat shiitake mycelium fermented product 7.24 27.84 35.08 Hwangchil Oat Shiitake Mycelium Fermented Product 6.16 26.27 32.43 Hwangchil Loquat Oat Shiitake Mycelium Fermented Product 8.35 29.84 38.19 Fermented product of turmeric oat shiitake mycelium 11.21 31.78 42.99

Table 21 shows the contents of vitamins B1 and B2 in the dog prototype to which the oriental medicinal herb extract mycelium fermented product was added. The content of vitamins B1 and B2 in the previously marketed dog treats was 2.21 and 11.93 mg/kg, respectively. The contents of vitamin B1 and B2 in the dog prototype were 8.41 to 10.26 and 23.11 to 33.42 mg/kg, respectively, which showed a significant increase compared to the existing commercial products. Vitamin B1 content is 10.26 mg/kg, containing 50 g of duck tenderloin, 35 g of fermented Hwangchil loquat oat shiitake mycelium, 5 g of Wongogogo extract, 2 g of Astragalus extract, 3 g of Baekbokryeong extract, 3 g of bamboo leaf extract, and 2 g of baekchul extract. The highest level was shown in Dog 3 prepared with a mixed ratio, and the highest level of vitamin B2 in Dog 3 was 33.42 mg/kg. Based on this data, the final dog prototype was selected as Dog 3. Among water-soluble vitamins, vitamin B1 is essential in food metabolism and is involved in energy generation and nerve impulse transmission for cell function. And it is effective in strengthening the resistance to disease.

Dog health snack Vitamin B1, B2 content (mg/kg) sample vitamin B1 vitamin B2 Sum Comparative example (commercial product) 2.21 11.93 14.14 dog 1 8.41 29.01 37.42 dog 2 9.41 30.04 39.45 dog 3 10.26 33.42 43.68 dog 4 9.53 28.31 37.84 dog 5 8.67 23.11 31.78

Table 22 shows the contents of vitamins B1 and B2 in the cat prototype to which the oriental medicinal herb extract mycelium fermented product was added. The content of vitamins B1 and B2 in previously marketed cat snack products was 3.16 and 9.34 mg/kg, respectively. The content of vitamins B1 and B2 in the cat prototype was 7.26 to 10.07 and 28.11 to 34.42 mg/kg, respectively, indicating a significant increase compared to the existing commercial products. The vitamin B1 content is 10.07 mg/kg, with a mixing ratio of 60 g of tuna, 25 g of fermented turmeric oat shiitake mycelium, 3 g of Wongogogo extract, 2 g of Astragalus extract, 3 g of baekbokryeong extract, 2 g of bamboo leaf extract, and 5 g of baekchul extract. Manufactured cat 4 showed the highest level, and vitamin B2 also showed the highest level at 34.42 mg/kg in cat 4. Based on this data, the prototype for the final cat was selected as Cat 4.

Cat snack vitamin B1, B2 content (mg/kg) sample vitamin B1 vitamin B2 Sum Comparative example (commercial product) 3.16 9.34 12.5 cat 1 7.26 31.34 38.6 cat 2 8.34 29.34 37.68 cat 3 8.26 31.41 39.67 cat 4 10.07 34.42 44.49 cat 5 9.01 28.11 37.12

Example 5. Inorganic component content

The content of calcium and phosphorus in each herbal medicine extract mycelium fermented product is shown in Table 23. The calcium content of the fermented product ranged from 160.16 to 450.21 mg/kg, showing a difference according to the herbal medicines. Among them, the fermented loquat oat shiitake mycelium showed significantly the highest content, followed by the Hwangchil loquat oat shiitake mycelium fermented product and the turmeric oat shiitake mycelium fermented product. The total vitamin phosphorus content of the mycelium fermented product according to the herbal extracts was in the range of 98.65~257.96 mg/kg, and, like the calcium content, the highest content was shown in the fermented loquat oat shiitake mycelium.

Fermented product calcium and phosphorus content (mg/kg) sample calcium sign ratio Loquat Oat Shiitake Mycelium Fermented Product 450.21 257.96 1:0.57 Hwangchil Oat Shiitake Mycelium Fermented Product 160.16 98.65 1:0.62 Hwangchil Loquat Oat Shiitake Mycelium Fermented Product 380.35 246.80 1:0.64 Fermented product of turmeric oat shiitake mycelium 302.38 202.88 1:0.67

Table 24 shows the calcium and phosphorus content of the dog prototype to which the oriental medicinal herb extract mycelium fermented product was added. The calcium and phosphorus content of the previously marketed dog treat products were 12.12 and 243.70 mg/kg, respectively, and the ratio was 1:2.02. The calcium and phosphorus contents of the dog samples using the fermented oriental herbal extract mycelium were 231.41~293.41 and 159.46~198.24 mg/kg, respectively, which showed no significant difference compared to the existing commercial products, but showed a significant difference in the ratio of calcium and phosphorus. The calcium content was 293.41 mg/kg, which was the highest in Dog 3, and the phosphorus content was the highest in Dog 1, at 198.24 mg/kg. Since the ratio of calcium and phosphorus is important for the absorption of calcium in the body, in order to increase the nutritional value, it is necessary to increase the content of calcium and phosphorus while maintaining a ratio suitable for calcium absorption (calcium/phosphorus = 2~1). Phosphorus is contained in many foods, so there is rarely a deficiency, but if the intake of phosphorus is too high, the absorption of calcium is inhibited and the loss of calcium increases. It was selected because it minimizes loss and can ingest a large amount of calcium.

Dog health snack calcium, phosphorus content (mg/kg) sample calcium sign ratio Comparative example (commercial product) 120.12 243.70 1:2.02 dog 1 270.31 198.24 1:0.73 dog 2 280.12 176.34 1:0.63 dog 3 293.41 168.21 1:0.57 dog 4 231.41 159.46 1:0.69 dog 5 241.21 182.32 1:0.76

Table 25 shows the calcium and phosphorus contents of the cat samples to which the oriental medicine shiitake mycelium fermented product was added. The calcium and phosphorus content of the previously marketed cat snack products were 124.86 and 231.46 mg/kg, respectively, and the ratio was converted to 1:1.85. The calcium and phosphorus contents of the cat samples using the herbal extract mycelium fermented product were 198.72~230.12 and 150.39~183.71 mg/kg, respectively, which showed no significant difference compared to the existing commercial products, but showed a significant difference in the ratio of calcium and phosphorus. The calcium content was 225.26 mg/kg, which was the highest in Cat 4, and the phosphorus content was the highest in Cat 1, 183.71 mg/kg. Among the following prototypes, the calcium and phosphorus ratio of 1:0.75 was selected because cat 4 minimizes calcium loss and can ingest a large amount of calcium when ingested by cats.

Cat snack calcium and phosphorus content (mg/kg) sample calcium sign ratio Comparative example (commercial product) 124.86 231.46 1:1.85 cat 1 230.12 183.71 1:0.80 cat 2 224.86 172.43 1:0.77 cat 3 210.71 168.34 1:0.80 cat 4 225.26 169.26 1:0.75 cat 5 198.72 150.39 1:0.76

Example 6. Heavy metal analysis result

As a result of heavy metal analysis, it was confirmed that heavy metals harmful to the health of dogs and cats were not detected (Table 26).

Fermented products and companion animal health snacks heavy metals sample Heavy metal (mg%) Pb CD Loquat oat shiitake mycelium fermented product N.D. N.D. Hwangchil oat shiitake mycelium fermented product N.D. N.D. Hwangchil Loquat Oat Shiitake Mycelium Fermented Product N.D. N.D. Fermented product of turmeric oat shiitake mycelium N.D. N.D. dog healthy snack ND ^* N.D. Cat Healthy Snacks N.D. N.D.

^* ND: not detected

Example 7. Cytotoxicity analysis results for Raw 264.7

1) Results of cytotoxicity analysis of herbal extracts

In order to confirm the cytotoxicity according to the concentration of the herbal extract, MTT assay was performed, and the results are shown in FIG. 13 . Loquat extract and Hwangchil extract showed toxicity at a concentration of 3000 μg/mL, respectively, with cell viability of 69.9% and 61.2%, and turmeric extract was toxic at a concentration of 2000 μg/mL with a cell viability of 69.5%. Therefore, it was finally set so that the loquat extract and hwangchil extract were added to the fermented herbal medicine extract at a concentration of 2000 μg/mL and the turmeric extract at a concentration of 1000 μg/mL.

2) Results of cytotoxicity analysis of fermented extracts of oriental medicine mycelia

MTT assay was performed to confirm the cytotoxicity of the fermented herbal extract mycelium against RAW264.7 cells, which are mouse macrophages. Cells for RAW264.7 cell of oat shiitake mycelium fermented product, turmeric oat shiitake mycelium fermented product, loquat oat shiitake mycelium fermented product, hwangchil oat shiitake mycelium fermented product, hwangchil loquat oat shiitake mycelium fermented product, cat health snack, dog health snack The toxicity analysis results are shown in FIG. 14 . As a result of confirming cytotoxicity at concentrations of 100, 500, and 1000 μg/mL, respectively, cytotoxicity was not observed in all samples and concentrations.

Example 8. NO (nitric oxide) production inhibitory effect analysis result

NO generated in the inflammatory state is known to play an important role in the human immune inflammatory response, such as promoting inflammatory reactions such as vascular permeability and edema, as well as promoting biosynthesis of inflammatory mediators. The results of the analysis of NO inhibitory effect of oat shiitake mycelium fermented product, turmeric oat shiitake mycelium fermented product, loquat oat shiitake mycelium fermented product, hwangchil oat shiitake mycelium fermented product, Hwangchil loquat oat shiitake mycelium fermented product, cat health snack, and dog health snack Same as 15. When the NO production rate of the LPS-treated group is 100%, at a concentration of 1000 μg/mL, fermented oat shiitake mycelium is 85.2%, turmeric oat shiitake mycelium fermented product is 61.4%, loquat oat shiitake mycelium fermented product is 88.2%, and hwangchil oats The fermented shiitake mycelium produced 84.7%, Hwangchil loquat oat shiitake mycelium fermented product 68.8%, cat healthy snack 78.3%, and dog health snack 81.6% NO production. It was found that the NO production rate was low in the order of loquat oat shiitake mycelium fermented product, cat healthy snack, and dog health snack.

Example 9. Measurement of inflammatory cytokine production

1) GM-CSF inhibitory effect analysis result

Analysis of GM-CSF inhibitory effect of oat shiitake mycelium fermented product, turmeric oat shiitake mycelium fermented product, loquat oat shiitake mycelium fermented product, hwangchil oat shiitake mycelium fermented product, hwangchil loquat oat shiitake mycelium fermented product, cat health snack, and dog health snack is shown in FIG. 16 . When the NO production rate of the LPS-treated group is 100%, at a concentration of 500 μg/mL, fermented oat shiitake mycelium is 96.2%, turmeric oat shiitake mycelium fermented product is 56.1%, loquat oat shiitake mycelium fermented product is 98.1%, and Hwangchil oats The fermented shiitake mycelium produced 89.3%, the fermented product of Hwangchil loquat oat shiitake mycelium 79.5%, the cat healthy snack 83.1%, and the dog health snack 86.1% GM-CSF production rate.

2) IL-1β inhibitory effect analysis result

The main function of IL-1β (Interleukin-1β) is as a mediator of the host inflammatory response to infection or other stimuli, similar to TNF. IL-1β is a relatively diverse cell such as monocytes, macrophages, keratinocytes, NK cells, T cells, B cells, endothelial cells, mast cells, neutrophils, fibroblasts, and neurons through stimulation of various production inducers including microorganisms. are being created in Its functions are also diverse, so it plays a major role in T cells, B cells, monocytes, macrophages, and NK cells. Analysis of IL-1β inhibitory effect of oat shiitake mycelium ferment, turmeric oat shiitake mycelium ferment, loquat oat shiitake mycelium ferment, hwangchil oat shiitake mycelium ferment, hwangchil loquat oat shiitake mycelium fermented product, cat health snack, and dog health snack is shown in FIG. 17 . When the NO production rate of the LPS-treated group is 100%, at a concentration of 500 μg/mL, fermented oat shiitake mycelium was 87.2%, turmeric oat shiitake mycelium fermented product was 35.9%, loquat oat shiitake mycelium fermented product was 85.5%, and Hwangchil oat The production rate of shiitake mycelium was 80.8%, the fermented product of Hwangchil loquat oat shiitake mycelium 40.7%, the healthy snack for cats 55.5%, and the healthy snack for dogs 60.7% of IL-1β production.

Korea Institute of Bioscience and Biotechnology KCTC18874P 20201127

Claims (7)

(1) Roasting loquat, adding water to the pulverized loquat for extraction, concentration and freeze-drying to prepare loquat extract powder;
(2) extracting by adding water to the pulverized hwangchil after roasting hwangchil, and concentrating and freeze-drying to prepare hwangchil extract powder;
(3) After adding water to a mixture of the loquat extract powder prepared in step (1) and the hwangchil extract powder prepared in step (2), oats are immersed in it, taken out and sterilized, and shiitake mycelium ( Lentinula edodes (Berk.)) inoculated and cultured, dried and pulverized to prepare a fermented product of Hwangchil loquat oat shiitake mycelium;
(4) extracting by adding water to the original target height and then concentrating to prepare an original target height extract;
(5) roasting astragalus and then adding water to the pulverized astragalus for extraction and concentration to prepare an astragalus extract;
(6) roasting bokryeong, adding water to the pulverized bokryeong, extracting it, and then concentrating to prepare a bokryeong extract;
(7) extracting by adding water to the pulverized bamboo leaves and then concentrating to prepare a bamboo leaf extract;
(8) roasting the baekchul and then adding water to the pulverized baekchul for extraction and concentration to prepare a baekchul extract; and
(9) Hwangchil loquat oat shiitake mycelium fermented product in step (3) for meat, Wongogogo extract prepared in step (4), Astragalus extract prepared in step (5), step (6) A method for producing a snack for companion animals, comprising adding and mixing the prepared bokryeong extract, the bamboo leaf extract prepared in the step (7), and the baekchul extract prepared in the step (8). The method of claim 1, wherein the shiitake mycelium ( Lentinula edodes (Berk.)) is a shiitake mycelium ( Lentinula edodes (Berk.)) JMIF-001 (KCTC18874P). The method of claim 1, wherein the companion animal is a dog or a cat. 3. The method of claim 2,
(1) Roasting loquats at 90-110° C. for 2-4 minutes, adding water to the pulverized loquats, extracting them at 70-90° C. for 8-12 hours, concentrating and freeze-drying to prepare loquat extract powder;
(2) roasting hwangchil at 140-160° C. for 8-12 minutes, adding water to the pulverized hwangchil, extracting it at 80-90° C. for 7-9 hours, and concentrating and freeze-drying to prepare hwangchil extract powder;
(3) After adding 1.4~1.6 L of water to a mixture of 12~18 g of loquat extract powder prepared in step (1) and 12~18 g of hwangchil extract powder prepared in step (2), here After immersing oats for 50~70 minutes, take them out and sterilize them, inoculate with shiitake mycelium ( Lentinula edodes (Berk.)) JMIF-001(KCTC18874P) and incubate at 22~26℃ for 3~5 weeks, then dry and pulverize to prepare a fermented product of hwangchil loquat oat shiitake mycelium;
(4) adding 80~120 L of water to 8~12 kg of original target height, extracting at 70~80℃ for 7~9 hours, and then concentrating to prepare an original target high extract;
(5) Roast 4-6 kg of astragalus at 140-160℃ for 3-10 minutes, add 45-55 L of water to the pulverized astragalus, extract at 70-90℃ for 10-14 hours, and then concentrate to prepare astragalus extract to do;
(6) After roasting 4~6 kg of bokryeong at 110~130℃ for 1~2 minutes, adding 30~40L of water to the pulverized bokryeong, extracting it at 90~100℃ for 10~14 hours, and concentrating to prepare bokryeong extract to do;
(7) adding 40-60 L of water to 4-6 kg of crushed bamboo leaves, extracting at 70-90° C. for 10-14 hours, and concentrating to prepare a bamboo leaf extract;
(8) After roasting 4-6 kg of baekchul at 140-160℃ for 2-4 minutes, adding 35-45 L of water to the pulverized baekchul, extracting at 80-100℃ for 8-12 hours, and concentrating to prepare a baekchul extract to do; and
(9) Hwangchil loquat oat shiitake mycelium fermented product in step (3) for meat, Wongogogo extract prepared in step (4), Astragalus extract prepared in step (5), step (6) A method for producing a snack for companion animals, comprising adding and mixing the prepared bokryeong extract, the bamboo leaf extract prepared in the step (7), and the baekchul extract prepared in the step (8). The method of claim 4, wherein the mixing in step (9) is 45-55 g of meat, 30-40 g of fermented Hwangchil loquat oat shiitake mycelium, 4-6 g of Wongogogo extract, 1.5-2.5 g of Astragalus extract, 2.5 g of Baekbokryeong extract. ~3.5 g, a method of manufacturing a snack for companion animals, characterized in that the mixture of 2.5-3.5 g of bamboo leaf extract and 1.5-2.5 g of baekchul extract. 5. The method of claim 4, wherein the mixing in step (9) is 55 to 65 g of meat, 20-30 g of fermented Hwangchil loquat oat shiitake mycelium, 2.5-3.5 g of Wongogogo extract, 1.5-2.5 g of Astragalus extract, 2.5 g of Baekbokryeong extract -3.5 g, 1.5-2.5 g of bamboo leaf extract, and 4.5-5.5 g of baekchul extract. A snack for companion animals manufactured by the method of any one of claims 1 to 6.

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