Method for producing immunoactivator for pets

Abstract

To provide a method for producing an immunoactivator for pets, which enables efficient and low-cost production from bamboo as the starting material.SOLUTION: A method for producing an immunoactivator for pets comprises: a pulverization step of pulverizing bamboo to provide bamboo powder; a fermentation step of fermenting the bamboo powder derived from the pulverization step by means of bamboo-derived lactic acid bacteria to provide a fermented bamboo product; an extraction step of subjecting the fermented bamboo product derived from the fermentation step to hot water extraction to provide a bamboo fermentation extract; a filtration step of filtering the bamboo fermentation extract derived from the extraction step; and a drying step of drying the residue formed in the filtration step.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a method for producing an immunopotentiator for pets, which is produced by subjecting a bamboo fermented product obtained by lactic acid fermentation of bamboo to hot water extraction treatment.

Background Art

[0002] Bamboo is a grass plant that is mainly found in Asian regions including Japan. Since it grows quickly and is easy to harvest, it has been widely used as a material for building materials, handicrafts, and daily necessities since ancient times. However, due to the emergence of chemical materials such as synthetic resins, the use of bamboo has decreased, and there has been a problem that unharvested bamboo forests erode mountains and farmland. In addition, even in managed bamboo forests, there has been a problem of high disposal costs because there are no takers for the harvested bamboo.

[0003] Therefore, in order to effectively utilize bamboo, research and development on new uses of bamboo are underway. For example, Patent Document 1 describes a method of pulverizing bamboo to produce bamboo powder, fermenting this bamboo powder with lactic acid bacteria to produce a bamboo fermented product, subjecting this bamboo fermented product to hot water extraction to produce an extract, and producing the filtrate after the hot water extraction treatment as a bamboo fermented extract.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] According to the production method described in Patent Document 1, a bamboo fermented extract having immunopotentiating activity can be produced. This bamboo fermented extract is used as an immunopotentiator for humans and is produced from the filtrate obtained by hot water extraction of the bamboo fermented product and filtering the extract. Therefore, it has a problem that the production efficiency from raw materials is low and the production cost is relatively high.

[0006] The present invention solves the above-mentioned problems, and its object is to provide a method for manufacturing an immunostimulant for pets that can be efficiently and inexpensively manufactured from bamboo as a raw material.

Means for Solving the Problems

[0007] According to the present invention, a method for manufacturing an immunostimulant for pets includes a pulverizing step of pulverizing bamboo to obtain bamboo powder, a fermenting step of fermenting the bamboo powder obtained in the pulverizing step with lactic acid bacteria derived from bamboo to obtain a bamboo fermented product, an extracting step of extracting the bamboo fermented product obtained in the fermenting step with hot water to obtain a bamboo fermented extract, a filtering step of filtering the bamboo fermented extract obtained in the extracting step, and a drying step of drying the residue generated in the filtering step.

[0008] According to the present invention, the immunostimulant for pets is manufactured by drying the residue generated when filtering the bamboo fermented extract obtained by hot water extraction of the bamboo fermented product. In this way, since the residue obtained in large quantities by filtering the bamboo fermented extract is used, and furthermore, since the residue generated by filtering the bamboo fermented extract, which was conventionally discarded, is utilized, the immunostimulant for pets can be efficiently manufactured from bamboo as a raw material. This leads to the ability to provide a sustainable immunostimulant for pets. Moreover, since the discarded residue is utilized, the immunostimulant for pets can be manufactured inexpensively.

[0009] Also, by using the bamboo powder obtained by pulverizing bamboo as a fermentation raw material, the surface area in contact with the bacterial cells increases, and sugars, amino acids, vitamins, etc. contained inside the cells are eluted into the bamboo powder due to pulverization. In addition, the lactic acid bacteria that lived in the fine honeycomb structure-like soft tissue, etc. are also dispersed in the bamboo powder. Therefore, the lactic acid bacteria grow efficiently and fermentation is promoted. And by extracting this bamboo fermented product with hot water, a bamboo fermented extract having an immunostimulating effect is obtained, so it has high safety and can be applied with confidence.

[0010] Furthermore, since the immunostimulant for pets produced by the present invention has the effect of activating the innate immune function, when a pet ingests it as food, the innate immune function, that is, the rapid infection defense against viruses, bacteria, etc. and the attack against malignant tumors, etc. can be activated and enhanced through the intestinal immune mechanism.

[0011] Preferably, the filtration step is a step of filtering the bamboo fermentation extract with a non-woven fabric to extract the residue, or centrifuging the bamboo fermentation extract to extract the residue.

[0012] It is also preferable that the fermentation step uses a culture containing lactic acid bacteria of the genus Weissella as a starter.

[0013] It is also preferable that the fermentation step is carried out under low vacuum or medium vacuum conditions, or in an inert gas atmosphere such as nitrogen gas.

[0014] It is also preferable that the hot water extraction in the extraction step is carried out under the conditions of 80°C to 130°C and 0.1 MPa to 0.3 MPa. These are the temperature and pressure conditions for performing suitable hot water extraction.

[0015] It is also preferable that the bamboo consists only of the branches and leaves of the bamboo. By using only the branches and leaves of the bamboo without using the bamboo trunk, the bamboo powder becomes more soluble in water.

Effects of the Invention

[0016] According to the present invention, since a large amount of residue obtained by filtering the bamboo fermentation extract is used, and furthermore, since the residue generated by filtering the bamboo fermentation extract that was conventionally discarded is utilized, the immunostimulant for pets can be efficiently produced from bamboo as a raw material. This leads to the ability to provide a sustainable immunostimulant for pets. Moreover, since the discarded residue is utilized, the immunostimulant for pets can be manufactured at low cost.

Brief Description of the Drawings

[0017]

Figure 1

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Embodiments for Carrying Out the Invention

[0018] FIG. 1 schematically shows a method for producing an immunostimulant for pets in one embodiment of the present invention.

[0019] As shown in the figure, the method for producing the immunostimulant for pets in this embodiment mainly consists of a step S1 of preparing bamboo as a raw material, a grinding step S2 of grinding the bamboo into bamboo powder, a fermentation step S3 of fermenting the bamboo powder with lactic acid bacteria to obtain a bamboo fermented product, a hot water extraction step S4 of hot water extracting the bamboo fermented product, a filtration step S5 of the hot water extract, and a drying step S6 of the residue remaining by filtration. Hereinafter, each of these steps will be described in detail.

[0020] (Preparation Step S1) In the preparation step S1, bamboo as a raw material is prepared. The bamboo used as the raw material for the bamboo fermentation extract according to the present embodiment generally widely includes plants of the Poaceae family called bamboo, sasaya, and bamboo. Specifically, although not particularly limited, examples include Phyllostachys heterocycla f.pubescence, Phyllostachys bambusoides, Phyllostachys nigra, Tetragonocalamus quadrangularis, Pleioblastus Simonii, Pseudosasa japonica, Sasa veitchii, etc. In particular, Phyllostachys heterocycla f.pubescence is preferably used. Since bamboo can be used as it is, except for washing the soil and the like attached to the bamboo, no pretreatment or the like is performed, and the cut bamboo can be directly used as the raw material for the bamboo fermentation extract of the present embodiment. By pulverizing fresh bamboo into powder or cutting it into chips, lactic acid bacteria existing inside the bamboo stem, particularly in the fine honeycomb-like soft tissue, are exposed and dispersed, and fermentation by lactic acid bacteria derived from bamboo proceeds. Among the bamboo, it is particularly preferable to use only the branches and leaves because they are easily soluble in water.

[0021] (Pulverization step S2) In the pulverization step S2, this bamboo is pulverized to obtain bamboo powder. As a method for pulverizing bamboo, any method may be used as long as the bamboo can be made into powder. When pulverizing the stem part other than the branches and leaves of bamboo, for example, a bamboo pulverizing device equipped with a rotary blade that operates when the cylindrical end face of fresh bamboo abuts can be used. In this type of bamboo pulverizing device, the fresh bamboo is pulverized into bamboo powder of a predetermined size by continuously cutting the cylindrical end face of the fresh bamboo in a direction substantially parallel to the cylindrical end face. By pulverizing the bamboo by shaving off the cylindrical end face of the fresh bamboo with a rotary blade, lactic acid bacteria existing in large numbers in the soft tissue and vascular bundle sheath of the bamboo stem are exposed, and fermentation in the subsequent fermentation step S3 is promoted. Further, by pulverizing the bamboo, the cell wall is broken, and sugars such as glucose, amino acids, and vitamins contained in the cells are also dispersed in the powder, and the nutritional requirements of lactic acid bacteria derived from bamboo in the fermentation step S3 are satisfied, and fermentation proceeds efficiently.

[0022] Although the particle size of the bamboo powder is not particularly limited, from the viewpoints that it becomes difficult to handle if it is too fine and the fermentation rate decreases if it is too large, it is preferably about 50 μm to 1 mm in particle size, and more preferably 200 μm to 600 μm in particle size. Also, it is preferable to introduce the bamboo powder obtained by the pulverization treatment into a metal removal device and adsorb and remove foreign substances due to chipping of the rotary blade of the bamboo powder pulverizing device. Further, the bamboo powder obtained by the pulverization treatment may be introduced into a sieving device to align the particle size range of the obtained bamboo powder or to remove uncut pieces that were not pulverized in the pulverization step S2.

[0023] (Fermentation step S3) Next, the fermentation step S3 will be described. The fermentation of the bamboo powder is carried out by lactic acid bacteria derived from bamboo. Specifically, the bamboo powder obtained by the pulverization step S2 is packed in a fermentation container, for example, a fermentation bag such as a transparent resin bag, in predetermined amounts, and after removing as much air as possible inside the fermentation bag, the mouth side of the fermentation bag is sealed to make the inside of the container in a substantially anaerobic state. This fermentation bag is covered with a light-shielding sheet such as a mat, and left standing in an environment of 20 to 30 °C for about 5 to 20 days, whereby a lactic acid ferment of the bamboo powder can be obtained. The fermentation state can be grasped from the pH value of the bamboo powder. That is, as lactic acid fermentation progresses, lactic acid is generated in the bamboo powder and the pH of the bamboo powder becomes acidic. Therefore, after putting 40 g of bamboo powder into 100 mL of purified water and stirring for 5 minutes, leaving it standing for 2 minutes, and filtering with filter paper, when the pH of the filtrate becomes 4.3 or less, it can be judged that the fermentation has sufficiently progressed, and the fermentation step S3 can be terminated.

[0024] Regarding the bamboo fermented product obtained in the fermentation step S3, when microbial community analysis was performed by denaturing gradient gel electrophoresis (DGGE method), in the initial stage of fermentation, mainly lactic acid bacteria of the genera Leuconostoc and Weissella were observed, and it was confirmed that lactic acid bacteria of the genus Weissella were the main component of the microbial community in the fermented bamboo product after fermentation. From this, it is also preferable to use a culture containing lactic acid bacteria of the genus Weissella as a starter in the fermentation step S3. By doing so, the dominant species of the microbial community in the bamboo powder can be quickly changed to the genus Weissella to promote fermentation, and the fermentation days in the fermentation step S3 can be shortened by about 30%, and can be about 3 to 15 days. Also, although the microbial community and the number of microorganisms vary depending on temperature fluctuations, the water content and type of bamboo, etc., by using a starter, a stable bamboo fermented product can be obtained throughout the year. It is preferable to add a starter containing lactic acid bacteria of the genus Weissella cultured separately in a liquid medium so that it is 0.5 mL to 5 mL per 1 kg of bamboo powder.

[0025] Also, in the fermentation step S3, it is also preferable to use a bamboo fermented product obtained by previously fermenting bamboo powder as a starter. In this way, by providing a seed preparation step in advance and using the obtained bamboo fermented product as a starter, the dominant species of the genus Weissella contained in the bamboo fermented product promotes the fermentation of new bamboo powder that has just passed through the grinding step S2, and the fermentation days in the fermentation step S3 can be shortened by about 30%, and can be about 3 to 15 days. Also, by using a starter, a stable bamboo fermented product can be obtained regardless of the climate or the state of the bamboo. In this way, when using the pre-prepared bamboo fermented product as a starter, it is preferable to add it so that it is 5 g to 20 g per 1 kg of new bamboo powder. Furthermore, it is also effective to use a part of the bamboo fermented product obtained in this fermentation step S3 as a starter in the next fermentation step.

[0026] Furthermore, it is also preferable to conduct the fermentation in the fermentation step S3 under low vacuum or medium vacuum conditions, or in an inert gas atmosphere such as nitrogen gas. This can prevent the growth of yeasts such as Hansenula or Pichia in the bamboo powder and prevent the generation of ethyl acetate resulting from acetic acid and ethanol produced by the yeast.

[0027] (Hot water extraction step S4) In the hot water extraction step S4, the bamboo fermented product obtained in the fermentation step S3 is subjected to hot water extraction to obtain a bamboo fermented extract. Examples of the hot water extraction method include reflux heating extraction and pressurized hot water extraction using an autoclave device or the like. Specifically, for example, the bamboo fermented product and water are placed in a reaction vessel, and reflux heat treatment or the like is performed for a predetermined time to extract the bamboo fermented extract. It is preferable to add 4 to 15 liters of water, and more preferably 4 to 10 liters of water, per 1 kg of the bamboo fermented product. Although water is used as the extraction solvent, other components can also be contained within a range that does not interfere with the extraction efficiency and does not affect the action of the obtained bamboo fermented extract. The extraction temperature is preferably 80°C or higher, more preferably 80°C to 130°C, and particularly preferably 100°C to 121°C. The extraction is carried out under atmospheric pressure or under pressure, and as the pressurization conditions, 0.1 MPa to 0.5 MPa is preferable, and 0.1 MPa to 0.3 MPa is more preferable. The extraction time is preferably 10 minutes to 5 hours, and more preferably 20 minutes to 2 hours.

[0028] (Filtration step S5) In the filtration step S5, the bamboo fermented extract obtained in the hot water extraction step S4 is filtered through a cloth or cotton cloth to extract the residue, or this bamboo fermented extract is centrifuged to extract the residue. After the cloth filtration treatment, by squeezing approximately 231 kg of the bamboo powder containing moisture (15 coolings per week with the current equipment) with a manual compression squeezer, a residue with a weight of approximately 47 kg can be obtained. Note that the extract of the bamboo fermented extract obtained in this filtration step S5 is used for the production of an immunopotentiator for humans.

[0029] (Drying step S6) In the drying step S6, the residue obtained in the filtration step S5 is dried, whereby an immunostimulant for pets is obtained. More specifically, in this drying step S6, drying is performed at 35°C for 14 hours using a shelf dryer, whereby about 12 kg of residue is obtained. Thereafter, it is processed to about 35 mesh using a powder machine. Further thereafter, it is sterilized at 120°C for about 120 minutes using a dry heat sterilizer. According to the bacterial count inspection, with dry heat sterilization at 135°C for 43 minutes, the bacterial count is 7.9×10 2 , with dry heat sterilization at 120°C for 60 minutes, the bacterial count is 5.2×10 2 , with dry heat sterilization at 120°C for 90 minutes, the bacterial count is 0, therefore, the sterilization time before final shipment was set to 120°C for 120 minutes.

[0030] The immunostimulant for pets produced in this way has immunostimulatory activity. Immunostimulatory activity means an action of improving both the functions of innate immunity and acquired immunity. In particular, it has an action of activating the innate immune function and an action of inducing the production of IL-12 and IFN-γ. Thereby, the innate immune function, that is, actions such as rapid infection defense against pathogens and attack against malignant tumors can be activated and enhanced. It also has an action of activating the immune response when the immune function is reduced and an action of stably maintaining the immune function. In addition, since this immunostimulant for pets has an action of inducing the production of IL-12 and IFN-γ, it is expected to not only activate Th1 cells and enhance cell-mediated immunity, but also have an action of suppressing humoral immunity and allergic reactions due to the enhancement of cell-mediated immunity.

[0031] As described in detail above, according to this embodiment, the immunostimulant for pets is produced by drying the residue generated when filtering the bamboo fermentation extract obtained by hot water extraction of the bamboo fermented product. In this way, since the residue obtained in large quantities by filtering the bamboo fermentation extract is used, and furthermore, since the residue generated by filtering the bamboo fermentation extract, which was conventionally discarded, is utilized, the immunostimulant for pets can be efficiently produced from bamboo as the raw material. This leads to the ability to provide a sustainable immunostimulant for pets. Moreover, the immunostimulant for pets can be produced at low cost.

[0032] Examples will be described below. However, the following examples relate to immunostimulants for humans, not immunostimulants for pets. The immune function of the immunostimulant for pets can be inferred from the immune function of the immunostimulant for humans.

Example

[0033] [Example 1] 1. Production of the immunostimulant for humans (1) Moso bamboo with a stem height of about 4 m was harvested, the leaf part was cut off with a saw to obtain only the stem part, and then it was washed with water to remove mud, dirt, etc. About 4 m of the stem part of this Moso bamboo was pulverized using a bamboo pulverizer (Bump - Mill, manufactured by Takaku Alloy Co., Ltd.) to obtain about 10 kg of bamboo powder with a particle size of 500 μm or less. Next, 10 kg of the pulverized bamboo powder was put into a resin bag with a capacity of 15 kg, the outer periphery of the resin bag was pressed to remove as much air inside the bag as possible, and then it was sealed. The resin bag was covered with a light - shielding sheet and left standing in an environment at 25°C for 14 days. When a part of the bamboo fermented product after 14 days was taken and microbial community analysis was performed by denaturing gradient gel electrophoresis (DGGE method), it was confirmed that lactic acid bacteria of the genus Weissella (Weissella cibaria or confusa) were the main components of the microbial community. 1 kg of the obtained bamboo fermented product was put into a reaction vessel for reflux heating, 5 L of water was poured in, and hot water extraction was carried out by heating under reflux. The reaction time was set to 60 minutes. After the reaction was completed, the mixture in the reaction vessel was filtered, and the filtrate (about 4.5 L) was recovered as the extract. To 4.5 L of this extract, 6 L of a 15% ethanol aqueous solution was added to precipitate the insoluble components, and the precipitate was recovered. The recovered precipitate was freeze - dried to obtain about 30 g of an immunopotentiator (1) for humans.

[0034] [Example 2] 2. Production of immunopotentiator (2) for humans The moso bamboo with a stem height of about 4 m was harvested, the leaf part was cut off with a saw, and only the stem part was left. Then it was washed with water to remove mud, dirt, etc. About 4 m of the stem part of this moso bamboo was pulverized using a bamboo pulverizer (Bump - Mill, manufactured by Takaku Alloy Co., Ltd.) to obtain about 10 kg of bamboo powder with a particle size of 500 μm or less. To 10 kg of the bamboo powder, 100 g of the bamboo fermented product obtained in Example 1 was added and mixed well. Next, this bamboo powder was put into a resin bag with a capacity of 15 kg, the outer periphery of the resin bag was pressed to remove as much air inside the bag as possible, and then it was sealed. The resin bag was covered with a light - shielding sheet and left standing in an environment at 25 °C for 9 days. 1 kg of the obtained bamboo fermented product was put into a reaction vessel for reflux heating, 5 L of water was poured in, and hot - water extraction was carried out by heating under reflux. The reaction time was set to 60 minutes. After the reaction was completed, the mixture in the reaction vessel was filtered, and the filtrate (about 4.5 L) was recovered as the extract. To 4.5 L of this extract, 6 L of a 15% ethanol aqueous solution was added to precipitate the insoluble components, and the precipitate was recovered. The recovered precipitate was freeze - dried to obtain about 30 g of the immunopotentiator (2) for human use.

[0035] [Example 3] 3. Production of the immunopotentiator (3) for human use for the immunological function activity test The bamboo fermented product obtained in Example 1 was suspended in sterilized physiological saline (0.9% NaCl) to a sample concentration of 100 mg / mL. This suspension was subjected to hot - water extraction treatment of the bamboo fermented product at 121 °C for 20 minutes in an autoclave. The hot - water extract was centrifuged to recover the supernatant, and the immunopotentiator (3) for human use was obtained. This supernatant was used for the immunopotentiating test. Also, 1 mL of the recovered supernatant was subjected to centrifugal evaporation to dryness, and the concentration of the immunopotentiator (3) for human use in the supernatant was calculated from the obtained dry weight.

[0036] [Example 4] 4. Production of the immunopotentiator (4) for human use for the immunological function activity test The bamboo ferment obtained in Example 2 was suspended in sterilized physiological saline (0.9% NaCl) to a sample concentration of 100 mg / mL. This suspension was subjected to hot water extraction treatment of the bamboo ferment at 121°C for 20 minutes in an autoclave. The hot water extract was centrifuged to recover the supernatant, and the immunopotentiator (4) for humans was obtained. This supernatant was used for the immunopotentiation test. Also, 1 mL of the recovered supernatant was subjected to centrifugal evaporation to dryness, and the concentration of the immunopotentiator (4) for humans in the supernatant was calculated from the obtained dry weight.

[0037] [Example 5] [5. Immunological function activity test] As the immunological function activity test, a natural immunity promotion activity test using silkworm muscle contraction as an index was conducted. That is, in this test method, when a test sample is administered to the larvae of a holometabolous insect such as a silkworm, rapid muscle contraction occurs if a neurotransmitter is present in the test sample, but when a substance having a natural immunity function activation effect is administered to the larvae of a holometabolous insect, slow muscle contraction (tardive muscle contraction) that takes about 10 minutes to complete occurs based on the finding. The immunopotentiator (3) for humans obtained in Example 3 and the immunopotentiator (4) for humans obtained in Example 4 were used as test samples for the test.

[0038] The measurement of the slow muscle contraction of silkworms was performed as follows (see Hamamoto H., Kamimura M., Sekimizu K., J. Biol. Chem., 2008, 283(4), pp. 2185 - 91). 50 μL of the test sample was injected into the body cavity of the decapitated muscle specimens of 5-day-old silkworms. The body length of the silkworms was measured, and the maximum value (after about 10 minutes) of the muscle contraction value (Contraction Value, C value = ([length before contraction (before injection)] - [length after contraction (after injection)]) / [length before contraction]) was determined. The test sample was serially diluted to a predetermined concentration with 0.9% physiological saline as shown in Table 1, and the C value was measured for each dilution. When the C value = 0.15, the slow muscle contraction activity was defined as 1 unit. The amount of the test sample that gave a C value = 0.15 was determined from the obtained dose-response curve, and the activity per 1 mg of the test sample was calculated. Also, 50 μL of physiological saline was used as a negative control, and 0.2 mL of air was used as a positive control. The results are shown in Table 1 and Figure 2. Figure 2(a) shows the results of the immunopotentiator (3) for humans, and Figure 2(b) shows the results of the immunopotentiator (4) for humans.

[0039] [Table 1]

[0040] [Comparative Example 1] Production and immunological function activity test of immunopotentiator for humans for immunological function activity test 1 kg of the bamboo fermented product obtained in Example 1 was placed in a 10-L plastic container, 20 g of bamboo charcoal powder and 6 g of papain enzyme were added, and they were mixed well. Further, 8 L of water was poured, the lid was closed, and it was left standing at 25°C for 14 days. After 14 days, the mixture in the container was filtered, and the filtrate (about 8 L) was recovered as lactic acid-fermented bamboo liquid. 10 mL of this lactic acid-fermented bamboo liquid was taken, the pH was examined with pH test paper and neutralized with NaOH. Also, 1 mL of the lactic acid-fermented bamboo liquid was subjected to a centrifugal evaporator to be dried, and the concentration of the components contained in the lactic acid-fermented bamboo liquid was calculated from the obtained dry weight. An immunological function activity test similar to Example 5 was performed using this lactic acid-fermented bamboo liquid as the test sample. The results are shown in Table 2 and Figure 3.

[0041]

Table 2

[0042] [Example 6] 7. Production of Immunopotentiator (5) for Human Use 10 g of the bamboo fermented product obtained in Example 1 was suspended in 100 mL of sterilized physiological saline (0.9% NaCl). This suspension was subjected to hot water extraction treatment of the bamboo fermented product in an autoclave at 121 °C and 2 atm for 20 minutes. The hot water extract was centrifuged to recover the supernatant, which was then dried using a centrifugal evaporator to obtain 440 mg of the powdered immunopotentiator (5) for human use. In addition, when the amount of saccharides contained in the obtained immunopotentiator (5) for human use was quantified by the phenol-sulfuric acid method, it was found that 7% by weight of the immunopotentiator (5) for human use was saccharides (see Table 4).

[0043] [Example 7] 8. Production of Immunopotentiator (6) for Human Use 10 g of the bamboo fermented product obtained in Example 1 was suspended in 100 mL of sterilized physiological saline (0.9% NaCl). This suspension was subjected to hot water extraction treatment of the bamboo fermented product in an autoclave at 121°C and 2 atm for 20 minutes. The hot water extract was centrifuged to recover the supernatant. 95% ethanol was added to the recovered supernatant, and ethanol was added until the final concentration of ethanol reached 68%. The generated precipitate was recovered and dried to obtain 27 mg of an immunopotentiator (6) for humans in powder form. Further, when the amount of saccharides contained in the obtained immunopotentiator (6) for humans was quantified by the phenol-sulfuric acid method, it was found that 52% by weight of the immunopotentiator (6) for humans was saccharides (see Table 4).

[0044] [Example 8] 9. Immunological function activity test for immunopotentiator (5) for humans and immunopotentiator (6) for humans

[0045] [Table 3] ​When measuring the immunological function activity of the human immunopotentiator obtained in Example 6 and Example 7, a dose-response curve as shown in Fig. 4 was observed, and it was found that this human immunopotentiator has natural immunity promoting activity. The value of the specific activity obtained from this dose-response curve was 7.3 units / mg for the human immunopotentiator (5), and 230 units / mg for the human immunopotentiator (6) that had undergone the ethanol precipitation formation process (see Table 4). From this, it was revealed that the bamboo fermentation extract obtained by hot water extraction of the bamboo fermented product has an immunopotentiating effect, that the components having the immunopotentiating effect can be concentrated or selectively obtained by ethanol precipitation, and that the human immunopotentiator (6) that had undergone the precipitation formation process exhibits a higher immunopotentiating effect. Also, as shown in Table 4 below, the content of saccharides in the human immunopotentiator (6) has increased to 52% by weight. Thus, since the specific activity also increases with the increase in the content of saccharides, it is considered that the polysaccharides contained in the bamboo fermentation extract are one of the active components having the immunopotentiating effect.

[0046] [Table 4]

[0047] [Example 9] 10. Production of the human immunopotentiator (7) 1 kg of the bamboo fermented product obtained in Example 1 was placed in a reaction vessel for reflux heating, 10 L of water was poured in, and hot water extraction was carried out at 100 °C by heating under reflux. The reaction time was 120 minutes. After completion of the reaction, the mixture in the reaction vessel was filtered, and about 9 L of the filtrate was recovered. This filtrate was concentrated to about 10 times to obtain a concentrated solution. To this concentrated solution, 95% ethanol was added, and ethanol was added until the final concentration of ethanol reached 70%. The precipitate formed by filtering this mixed solution was recovered and dried to obtain a powdered human immunopotentiator (7).

[0048] [Example 10] 11. Immunoresponse test using mouse spleen cells Spleen cells were collected from C57B6 mice, and the spleen cells were in RPMI-1640 liquid medium at 5×10 6It was adjusted to be present at [number of cells] / mL. As the test sample, the human immunopotentiator (7) produced in Example 9 was used. Also, since natural product-derived extracts often contain LPS, as a comparative sample, lipopolysaccharide (LPS) was used to confirm whether the production of cytokines by this human immunopotentiator was due to LPS. Spleen cells of C57B6 mice ([5×10 6 cells / mL) were added with the human immunopotentiator (7) so that the final concentrations were 2 μg / mL, 5 μg / mL, and 10 μg / mL, and cultured for 48 hours. Similarly, LPS was added to spleen cells of C57B6 mice ([5×10 6 cells / mL) so that the final concentration was 0.1 μg / mL, and cultured for 48 hours. After culturing for 48 hours, the production amounts of IL-12 (p70) and IFN-γ in the medium were measured by the ELISA method. The results of the IL-12 (p70) production amount are shown in Figure 5, and the results of the IFN-γ production amount are shown in Figure 6. As shown in Figures 5 and 6, it was found that this bamboo fermentation extract induced the production of IL-12 and IFN-γ. Also, since no production of either IL-12 or IFN-γ was observed from the spleen cells to which LPS was added, it was considered that the production of these cytokines by this bamboo fermentation extract was not due to LPS, but was an action by components other than LPS contained in the bamboo fermentation extract.

[0049] All of the above-described embodiments and examples are illustrative of the present invention and not restrictive, and the present invention can be implemented in various other modified and changed forms. Therefore, the scope of the present invention is defined only by the scope of the claims and its equivalent scope.

Claims

1. A method for producing an immunopotentiator for pets, comprising: a pulverizing step of pulverizing bamboo to obtain bamboo powder; a fermenting step of fermenting the bamboo powder obtained by the pulverizing step with lactic acid bacteria derived from bamboo to obtain a bamboo fermented product; an extracting step of extracting the bamboo fermented product obtained by the fermenting step with hot water to obtain a bamboo fermented extract; a filtering step of filtering the bamboo fermented extract obtained by the extracting step; and a drying step of drying the residue generated in the filtering step.

2. The method for producing an immunopotentiator for pets according to claim 1, wherein the filtering step is a step of filtering the bamboo fermented extract with a non-woven fabric to extract the residue, or a step of centrifuging the bamboo fermented extract to extract the residue.

3. The method for producing an immunopotentiator for pets according to claim 1, wherein the fermenting step uses a culture containing lactic acid bacteria of the genus Weissella as a starter.

4. The method for producing an immunopotentiator for pets according to claim 1, wherein the fermenting step is performed under low vacuum or medium vacuum conditions, or in an inert gas atmosphere such as nitrogen gas.

5. The method for producing an immunopotentiator for pets according to claim 1, wherein the hot water extraction in the extracting step is performed under the conditions of 80°C to 130°C and 0.1 MPa to 0.3 MPa.

6. The method for producing an immunopotentiator for pets according to claim 1, wherein the bamboo consists only of the branches and leaves of bamboo.

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