Preparation method and application of lucid ganoderma liquid fermentation product for animal feed

Abstract

The invention belongs to the technical field of animal feed processing, and discloses a preparation method of a ganoderma lucidum liquid fermentation product for animal feed, which comprises the following steps: S1, filling a fermentation tank with a proper amount of water, heating to a proper temperature, adding a proper amount of fermentation medium, fully stirring, uniformly mixing, supplementing water to a constant volume, and sealing; sterilizing and cooling; s2, 0.6%-1.2% of ganoderma lucidum fermentation strains are rapidly inoculated into the fermentation tank through the inoculation opening, and the inoculation opening is closed; and S3, performing continuous constant-temperature fermentation culture for 7-14 days under proper fermentation culture conditions to obtain the lucid ganoderma liquid fermentation product. The invention also discloses a ganoderma lucidum liquid fermentation product prepared by the preparation method as a calf feed additive. When the lucid ganoderma liquid fermentation product is used as a calf feed additive, the growth and development of calves can be promoted, the proliferation of blood immune cells and the expression of cell factors and immune globulins can be promoted, and the diversity of intestinal microorganisms can be improved.

Description

Technical Field

[0001] The invention relates to the technical field of animal feed processing, and in particular to a preparation method and application of a ganoderma lucidum liquid fermentation product for animal feed. Background Art

[0002] Ganoderma lucidum contains a variety of components, such as glycoproteins, polysaccharides, triterpenes, sesquiterpenes, steroids, alkaloids, benzopyrans, and benzoic acid compounds. It also contains some minerals, such as potassium, calcium, phosphorus, magnesium, selenium, iron, and zinc. Polysaccharides and triterpenes are the main components of Ganoderma lucidum extracts, and their anticancer effects are mainly attributed to these two components. Ganoderma lucidum polysaccharides are composed of (1→3), (1→6)-α / β-glucans, glycoproteins, and water-soluble heteropolysaccharides. Ganoderma lucidum polysaccharides can exert anticancer effects through multiple mechanisms, such as inhibiting tumor growth and metastasis, and enhancing the patient's immune function.

[0003] Wild Ganoderma has very high requirements for the growth environment, so the output of wild Ganoderma is seriously insufficient to meet the demand. Although artificial cultivation of Ganoderma has increased the output of Ganoderma, its output is unstable, the quality of products is uneven, and the cost is high. The Ganoderma mycelium produced by liquid fermentation contains effective ingredients such as Ganoderma polysaccharides that are not lower than the fruiting body. At the same time, the biologically active ingredients of the mycelium are easy to separate and apply, and the effect of gastrointestinal absorption is better than that of the fruiting body and spore powder.

[0004] Calf health is an important part of ensuring the healthy development of the cattle industry. There are many diseases that cause calf death, such as bacterial and viral diseases caused by low immunity, which ultimately lead to animal death. The use of antibiotics now brings negative problems. Antibiotic resistance has become one of the main threats to global public health. Reducing or even avoiding the abuse of antibiotics and regulating the use of antibiotics in animal husbandry is our development strategy.

[0005] To this end, the present invention proposes a feed additive composed of ganoderma lucidum liquid fermentation product capable of enhancing immunity to replace antibiotics. Summary of the invention

[0006] The present invention aims to provide a method for preparing liquid fermented ganoderma lucidum for animal feed. The feed additive composed of liquid fermented ganoderma lucidum is applied to animal feed, which can promote the growth and development of calves, promote the proliferation of blood immune cells, the expression of cytokines and immunoglobulins and improve the diversity of intestinal microorganisms; it is a green and safe feed additive that is non-toxic, has no side effects and does not pollute the environment.

[0007] In order to achieve the above object, the present invention provides the following technical solutions:

[0008] The present invention provides a method for preparing a liquid fermented product of Ganoderma lucidum for animal feed, comprising the following steps:

[0009] S1. Add an appropriate amount of water into the fermentation tank, heat it to boiling, release a small amount of hot water from the liquid outlet, add cold water to a suitable temperature, add an appropriate amount of fermentation medium, stir and mix thoroughly, add water to the fixed volume and then seal; sterilize and cool down;

[0010] S2, quickly inoculating 0.6%-1.2% of the fermented Ganoderma lucidum strain into the fermentation tank through the inoculation port, and closing the inoculation port;

[0011] S3, continuously fermenting and culturing at a constant temperature for 7-14 days under suitable fermentation and culture conditions to obtain a Ganoderma lucidum liquid fermentation product;

[0012] Preferably, in step S3, the constant temperature fermentation culture temperature of the Ganoderma lucidum liquid fermentation product is 28-31°C.

[0013] Preferably, in step S3, the Ganoderma lucidum liquid fermentation product needs to be ventilated during constant temperature fermentation, and the ventilation volume is 6-14 L·min -1 .

[0014] Preferably, in step S3, the pH of the constant temperature fermentation culture of the Ganoderma lucidum liquid fermentation product is 5-6.

[0015] Preferably, in step S1, the carbon source material in the fermentation medium is soluble starch.

[0016] Preferably, in step S1, the nitrogen source in the fermentation medium is soybean powder.

[0017] Preferably, in step S1, the formula of the fermentation medium is 2% soluble starch, 1.8% soybean powder, 0.3% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and 0.005% vitamin B1.

[0018] The invention also provides an application of the fragrant Ganoderma lucidum liquid fermentation product prepared by the method for preparing the Ganoderma lucidum liquid fermentation product for animal feed as a feed additive for livestock animals.

[0019] The present invention also provides a method for preparing a liquid fermented product of Ganoderma lucidum for animal feed, and uses the prepared liquid fermented product of Ganoderma lucidum as a feed additive for calves, wherein the amount added to each calf complete feed is 25 g.d per head of calf. -1 .

[0020] In summary, the present invention has the following beneficial effects:

[0021] The ganoderma liquid fermentation product provided by the invention is used as a feed additive for calves, which is a green and safe feed additive that is non-toxic, has no side effects and does not pollute the environment. It can promote the growth and development of calves, promote the proliferation of blood immune cells, the expression of cytokines and immunoglobulins, and improve the diversity of intestinal microorganisms. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a diagram showing the growth state of the Ganoderma lucidum block cultured on a plate in the present invention;

[0023] Figure 2 The effect of the culture temperature on the dry weight of Ganoderma lucidum mycelium in the present invention;

[0024] Figure 3 This is a graph showing the effect of ventilation volume on the dry weight of Ganoderma lucidum mycelium in the present invention;

[0025] Figure 4 This is a graph showing the effect of pH on the dry weight of Ganoderma lucidum mycelium in the present invention;

[0026] Figure 5 The figure shows the effect of adding the liquid fermented product of Ganoderma lucidum to animal feed on calf cytokines in the present invention (A is the effect on interleukin-2, B is the effect on tumor necrosis factor-a, C is the effect on immunoglobulin A, D is the effect on immunoglobulin G, and E is the effect on immunoglobulin M, where * represents a significant difference (P<0.05) and ** represents an extremely significant difference (P<0.01)). DETAILED DESCRIPTION

[0027] The following embodiments of the technical solution of the present invention are described in detail. The following embodiments are used to more clearly illustrate the technical solution of the present invention, and therefore, are only used as examples and cannot be used to limit the protection scope of the present invention.

[0028] Example

[0029] 1. Screening of Ganoderma lucidum fermentation strains

[0030] The fermentation strains were screened by two methods: plate solid culture method and liquid shake flask culture method. The plate solid culture method used the mycelium growth rate as the criterion, and the faster the growth rate, the better; the liquid shake flask culture method used the mycelium dry weight as the criterion, and the heavier the dry weight, the better.

[0031] Test strains: Yellow-brown Ganoderma lucidum (numbers HD002, HD006, HD008), Black Ganoderma lucidum (numbers CH403, CH406, CH407), Red Ganoderma lucidum (numbers CD102, CD104, CD107); the strains were preserved in this laboratory and donated by related enterprises.

[0032] (1) Plate culture selection method for Ganoderma lucidum strains

[0033] The collected Ganoderma lucidum strains were restored to room temperature for later use. The strains with a diameter of 0.5 cm were selected and placed on PDA solid medium. The growth of the colonies was observed at 28°C for 7 days and the mycelial growth rate was measured. The experimental results are shown in Tables 1 and Figure 1 .

[0034] Table 1 Growth rate of mycelium of nine species of Ganoderma lucidum

[0035]

[0036] According to Table 1 and Figure 1 The results show that the selection results of CH403, CH406, HD008 and HD006 Ganoderma lucidum blocks are better through the plate culture method.

[0037] (2) Method for selecting Ganoderma lucidum strains in liquid shake bottles

[0038] Take a Ganoderma lucidum block with a diameter of about 0.5 cm and add it to a shake flask. Set up 3 replicates for each strain and culture it in a shaking incubator at 28°C with a rotation speed of 160 r / min. Take it out after 7 days and measure the dry weight of the mycelium.

[0039] Method for determining the dry weight of mycelium: take a certain volume of fermentation liquid, centrifuge at 5000r / min for 30min, discard the supernatant, wash the precipitate with distilled water, centrifuge again, repeat 3 times, and dry the obtained precipitate at 60℃ to constant weight. The experimental results are shown in Table 2.

[0040] Table 2 Dry weight of mycelium of nine kinds of Ganoderma lucidum at 7 days

[0041]

[0042] According to the results in Table 2, the results of the shake flask selection method showed that CH403, HD006 and HD008 were the best Ganoderma lucidum clumps.

[0043] 2. Optimize fermentation medium

[0044] The selected strain CH403 was transferred to shake flask culture. The culture medium was optimized by single factor test method, with glucose, sucrose, maltose, and soluble starch as carbon sources, beef extract, peptone, yeast extract, and soybean powder as nitrogen sources, and single factor test was designed to screen out the most suitable carbon and nitrogen sources. The optimized culture medium was used in the subsequent fermentation tank, and the fermentation product obtained by the shake flask was the strain used in the fermentation tank.

[0045] The results of the single factor test of carbon and nitrogen sources of the culture medium obtained by single factor test optimization are shown in Table 3. After comparison, it was found that soybean powder was more suitable as the nitrogen source and soluble starch was more suitable as the carbon source.

[0046] Table 3 Results of single factor test on carbon and nitrogen sources in culture medium

[0047]

[0048] The final culture medium composition was 2% soluble starch, 1.8% soybean powder, 0.3% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and 0.005% vitamin B1.

[0049] 3. Optimize fermentation conditions

[0050] A 120L fermentation tank is filled with 80L of liquid and heated to boiling; a small amount of hot water is released from the liquid outlet, cold water is added to a suitable temperature, culture medium and defoaming agent are added, the mixture is fully stirred and mixed, the mixture is poured into the feed port, water is added to the 100L mark, and the port is sealed; the pressure value is controlled, and sterilization is carried out at 115°C for 60min; after the sterilization is completed, condensed water is introduced into the circulating insulation pipe port to cool down; several bottles of shake flask culture are taken, and the inoculation port is quickly connected to the fermentation tank by the fire ring inoculation method, and the inoculation port is closed; continuous constant temperature culture is carried out for 14d to obtain the Ganoderma lucidum liquid fermentation product.

[0051] Single factor experiment was used to optimize the liquid fermentation conditions. 120L fermentation tank was filled with 80L of liquid, and the dry weight of mycelium per unit volume was used as the judgment standard.

[0052] (1) Investigating the effect of inoculation amount on mycelium dry weight

[0053] The culture temperature was 28°C, pH was 7, and the ventilation volume was 10 L min -1 The inoculation amounts were set to 0.6%, 0.9% and 1.2% respectively. The fermentation lasted for 14 days to determine the optimal inoculation amount. The experimental results are shown in Table 4.

[0054] Table 4 Effect of inoculation amount on dry weight of Ganoderma lucidum mycelium

[0055]

[0056] From the results in Table 4, it can be seen that under the condition of 0.6% inoculation amount, the dry weight of mycelium from 0 to 14 days increased with the fermentation time, but the fermentation culture time was long and the maximum dry weight was low; the Ganoderma mycelium cultured under the conditions of two gradient inoculation amounts of 0.9% and 1.2% showed a trend of first increasing and then decreasing with the increase of time. After 10 days of cultivation, the dry weight of Ganoderma mycelium obtained with an inoculation amount of 0.9% was the largest, reaching 14.92 g / L.

[0057] (2) Investigating the effect of temperature on mycelium dry weight

[0058] pH is 7, ventilation volume is 10L·min -1 The inoculation amount was 0.6%, and the fermentation temperatures were set at 25°C, 28°C, and 31°C, respectively. The fermentation lasted for 14 days to determine the optimal culture temperature. The experimental results are shown in Figure 2 .

[0059] Depend on Figure 2It can be seen that under the culture temperature of 25℃, the dry weight of mycelium from 0 to 14 days showed an increasing trend with the increase of fermentation time, but the fermentation culture time was long and the maximum dry weight was low; the Ganoderma mycelium cultured under the two gradient culture temperature conditions of 28℃ and 31℃ showed a trend of first increasing and then decreasing with the increase of time. The Ganoderma mycelium obtained after 10 days of culture at the culture temperature of 28℃ had the largest dry weight, which could reach 13.86g / L.

[0060] (3) Investigating the effect of ventilation volume on mycelium dry weight

[0061] The culture temperature was 28°C, pH was 7, inoculation volume was 0.6%, and ventilation volumes were set at 6 L·min-1, 10 L·min-1, and 14 L·min-1, respectively. The fermentation lasted for 14 days to determine the optimal ventilation volume. The experimental results are shown in Figure 3 .

[0062] Depend on Figure 3 It can be seen that 6L·min -1 Under the condition of ventilation, the dry weight of mycelium increased with the fermentation time from 0 to 14 days, but the maximum dry weight was low when the fermentation time was long; 9 L·min -1 、12L·min -1 The mycelium of Ganoderma lucidum cultured under two gradient culture temperature conditions showed a trend of first increasing and then decreasing with time. The dry weight of Ganoderma lucidum mycelium obtained at 28℃ for 10 days was the largest, reaching 14.40g / L.

[0063] (4) Investigating the effect of pH on mycelium dry weight

[0064] The culture temperature was 28°C, the ventilation volume was 10 L·min-1, the inoculation amount was 0.6%, and the pH was set to 5, 5.5 and 6 respectively. The fermentation was carried out for 14 days to determine the optimal pH. The experimental results are shown in Figure 4 .

[0065] Depend on Figure 4 It can be seen that when the pH is 5.0, the mycelium dry weight is the smallest, which is 7.34 g / L; when the pH increases to 5.5, the mycelium dry weight increases to 7.59 g / L; when the pH reaches 6.0, the mycelium dry weight is the largest, which can reach 8.23 ​​g / L; when the pH is 6.5, the mycelium dry weight decreases to 8.01 g / L, indicating that the optimal culture pH is 6.5.

[0066] Therefore, the steps of preparing liquid Ganoderma lucidum fermentation product by using the above-mentioned preferred process parameters are as follows:

[0067] S1. Add 100L of water to the middle of the perspective mirror above the fermentation tank in a 120L fermentation tank and heat it to boiling; release a small amount of hot water from the liquid outlet, add cold water to a suitable temperature, add 2200g of fermentation medium and 35g of defoaming agent, stir and mix thoroughly, add water to the feed port to make the volume 100L and seal it; control the pressure value, sterilize at 115℃ for 60min; after the sterilization is completed, condensed water is introduced into the circulating insulation pipe port to cool down to 28℃;

[0068] The formula of the fermentation medium is 2% soluble starch, 1.8% soybean powder, 0.3% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and 0.005% vitamin B1.

[0069] S2. Take HD008 Ganoderma lucidum strains, and use the fire ring inoculation method to quickly inoculate 0.6%-1.2% of Ganoderma lucidum fermentation strains into the fermentation tank through the inoculation port, and close the inoculation port.

[0070] S3, at pH 6, ventilation volume 9 L min -1 The fermentation temperature was 28°C and the culture was continued at a constant temperature for 14 days to obtain the Ganoderma lucidum liquid fermentation product.

[0071] The Ganoderma lucidum liquid fermentation product prepared by the present invention was commissioned to the Kunming Edible Fungi Quality Supervision, Inspection and Testing Center of the All-China Federation of Supply and Marketing Cooperatives to use the "NY / T 1676-2023 Spectrophotometric method for the determination of crude polysaccharides in edible fungi" and "NY / T3676-2020 Spectrophotometric method for the determination of total triterpenes in Ganoderma lucidum" to detect the contents of crude polysaccharides and total triterpenes in the Ganoderma lucidum liquid fermentation product, and the test results are shown in Table 5.

[0072] Table 5 Results of the detection of crude polysaccharides and total triterpenes in the liquid fermentation product of Ganoderma lucidum

[0073]

[0074] Feeding test example

[0075] 1.1 Experimental animals and groups

[0076] 32 healthy Simmental weaned calves of 4-5 months old and weighing 175±5kg were randomly selected from Inner Mongolia Horqin Beef Cattle Seed Co., Ltd., with similar body conditions. The weaned calves were randomly divided into 4 groups, 8 replicates in each group, 1 calf in each replicate, the control group (Con group) was fed with a basic diet, and the experimental groups were supplemented with different doses of powdered Ganoderma lucidum liquid fermentation product of Example 1 on the basis of the basic diet, with the added doses of 3g.d-1, 5g.d-1, and 10g.d-1 (3g group, 5g group, and 10g group), the pre-feeding period was 15d, and the feeding period was 60d.

[0077] 1.2 Effects of Ganoderma lucidum liquid fermentation product on growth performance of beef calves

[0078] During the feeding period, the total feed weight of each calf was accurately weighed with an electronic scale every day to calculate its average daily feed intake (ADFI). The body size indicators measured included the initial body weight (IBW), final body weight (FBW), body height (BH) and body oblique length (BL) of the calf. BH and BL were measured with a measuring stick at the end of the feeding period to calculate the average daily weight gain (ADG). The results are shown in Table 6.

[0079] Table 6 Effects of Ganoderma lucidum liquid fermentation on growth performance of calves

[0080]

[0081] Note: In the same data, no letter or the same lowercase letter on the shoulder indicates no significance (P>0.05), while different lowercase letters indicate significant difference (P<0.05), and different uppercase letters indicate extremely significant difference (P<0.01), the same as in the following table.

[0082] As shown in Table 6, the addition of Ganoderma lucidum liquid fermentation products to the diet had no significant effect on FBW, ADG, ADFI and BL of calves (P>0.05). Compared with the Con group and the 5g group, the FBW and ADFI of the 3g group and the 10g group showed an upward trend, among which the 5g group had the lowest level; compared with the 5g group, the ADG and BL of the 3g group and the 10g group showed an upward trend, but the difference was not significant (P>0.05); compared with the Con group, the BH of the 10g group was significantly higher than that of the Con group (P<0.05), and compared with the three experimental groups, the BH of the 10g group was significantly higher than that of the 5g group (P<0.05). The above results show that the addition of 3g.d to the diet can improve the calf's FBW, ADG, ADFI and BL. -1 , 10g.d -1 The effect of Ganoderma lucidum liquid fermentation on the growth performance of calves is better than that of the 5g group.

[0083] 1.3 Effects of Ganoderma lucidum liquid fermentation products on blood parameters of beef calves

[0084] On the 60th day of the feeding period, blood was collected from the jugular vein of 5 calves randomly selected from each group, 10 mL of which was placed in an EDTA anticoagulation vacuum tube, and the number of lymphocytes (LYM), the number of neutrophils (GRAN), the median number of cells (MID), the hemoglobin concentration (HGB) and the hematocrit (HCT) were detected using a fully automatic animal blood routine tester; 10 mL was placed in a procoagulant vacuum tube, left to stand at room temperature for 15 min, centrifuged at 3000 rpm for 15 min, the upper serum was placed in a 1.5 mL EP tube, and stored in a -20 ° C refrigerator. The serum indicators interleukin-2 (IL-2), interleukin-4 (IL-4), tumor necrosis factor-α (TNF-α), immunoglobulin A (IgA), immunoglobulin G (IgG) and immunoglobulin M (IgM) were determined using an ELISA kit (purchased from Jiangsu Enzyme-Free Industry Co., Ltd.). The results are shown in Tables 7 and Figure 5 .

[0085] Table 7 Effects of Ganoderma lucidum liquid fermentation on calf whole blood immune indexes

[0086]

[0087] As shown in Table 7, the addition of Ganoderma liquid fermentation product to the diet had no significant effect on the whole blood GRAN, MID and HGB of calves (P>0.05); the LYM of calves in the 10g group was significantly higher than that in the Con group (P<0.05); the HCT of the 10g group was extremely significantly higher than that in the Con group (P<0.01), and significantly higher than that in the 3g group (P<0.05); the HGB of the 10g group and the 3g group was extremely significantly higher than that of the control group (P<0.01).

[0088] Depend on Figure 5 A and Figure 5 D shows that IL-2 and IgG in the 3g and 10g groups were significantly higher than those in the Con group (P<0.01), and the 10g group was significantly higher than the 3g group (P<0.01). Figure 5 B and Figure 5 C shows that TNF-α and IgA in the 10g group were significantly higher than those in the Con group, among which TNF-α in the 3g group was significantly higher than that in the Con group (P<0.05), and IgA in the 10g group was significantly higher than that in the 3g group (P<0.01). Figure 5 As far as we know, the IgM in the 3g group was extremely significantly higher than that in the Con group (P<0.01), and the IgM in the 10g group was significantly higher than that in the Con group (P<0.05).

[0089] In summary, feeding Ganoderma lucidum liquid fermentation product can improve the growth and development of calves and enhance the immunity of calves, and the added dose of 10g is better than 3g.

[0090] The above test results show that the Ganoderma lucidum liquid fermentation product provided by the present invention can be used as a feed additive for calves to improve the growth and development of calves and has the effect of enhancing the immunity of calves.

[0091] The above is only an embodiment of the present invention, and the common knowledge such as the known specific technical solutions or characteristics in the solution is not described in detail here. It should be pointed out that for those skilled in the art, several modifications and improvements can be made without departing from the technical solution of the present invention, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the specification can be used to interpret the content of the claims.

Claims

1. A method for preparing a liquid fermented product of Ganoderma lucidum for animal feed, characterized in that: The steps include: S1. Add an appropriate amount of water into the fermentation tank, heat it to boiling, release a small amount of hot water from the liquid outlet, add cold water to a suitable temperature, add an appropriate amount of fermentation medium, stir and mix thoroughly, add water to the fixed volume and seal it; sterilize and cool down; S2, quickly inoculating 0.6%-1.2% of the fermented Ganoderma lucidum strain into the fermentation tank through the inoculation port, and closing the inoculation port; S3. Continuously perform constant temperature fermentation culture for 7-14 days under suitable fermentation culture conditions to obtain Ganoderma lucidum liquid fermentation product.

2. The method for preparing a liquid fermented product of Ganoderma lucidum for animal feed according to claim 1, characterized in that: In step S3, the temperature of the constant temperature fermentation culture of the Ganoderma lucidum liquid fermentation product is 28-31°C.

3. The method for preparing a liquid fermented product of Ganoderma lucidum for animal feed according to claim 1, characterized in that: In step S3, the liquid fermented product of Ganoderma lucidum needs to be ventilated during constant temperature fermentation, and the ventilation volume is 6-14 L·min -1 .

4. The method for preparing a liquid fermented product of Ganoderma lucidum for animal feed according to claim 1, characterized in that: In step S3, the pH of the constant temperature fermentation culture of the Ganoderma lucidum liquid fermentation product is 5-6.

5. The method for preparing a liquid fermented product of Ganoderma lucidum for animal feed according to claim 1, characterized in that: In step S1, the carbon source material in the fermentation medium is soluble starch.

6. The method for preparing a liquid fermented product of Ganoderma lucidum for animal feed according to claim 5, characterized in that: In step S1, the nitrogen source in the fermentation medium is soybean powder.

7. The method for preparing a liquid fermented product of Ganoderma lucidum for animal feed according to claim 6, characterized in that: In step S1, the fermentation medium comprises 2% soluble starch, 1.8% soybean powder, 0.3% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and 0.005% vitamin B1.

8. Use of the Ganoderma lucidum liquid fermentation product prepared by the method for preparing Ganoderma lucidum liquid fermentation product for animal feed according to any one of claims 1 to 7, characterized in that: Used as feed additive for livestock.

9. The use according to claim 8, characterized in that: The liquid fermented product of Ganoderma lucidum is used as a feed additive for calves, and the amount added to each calf complete feed is 25g / d. -1 .

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