Preparation method of biological feed additive

[0024] Preparation Example 1

[0025] 1. Select three traditional Chinese medicines of pine needles, astragalus, and hawthorn to be mixed with Ginkgo biloba leaves for fermentation, and select the appropriate Chinese medicine and its ratio with Ginkgo biloba leaves and bran.

[0026] 2. The experiment adopts a two-factor completely randomized design, with a control, a total of 10 treatments, repeated 3 times, the experimental design is shown in Table 1.

[0027] Table 1 Two-factor completely random design table

[0028]

[0029] Table 2 Fermentation comparison of different Chinese medicine-ginkgo biloba combinations

[0030]

[0031] Note: Different lowercase letters indicate significant differences (P<0.05), and different capital letters indicate extremely significant differences (P<0.01).

[0032] 3. The test results show that the fermentation effect of using astragalus-ginkgo biloba-bran as the fermentation raw material is better (Table 2). The fermented product particles of pine needles-ginkgo biloba-bran are coarser and have a strong rosin taste. The yeast form is fuller, but the number is not large; the product particles after fermentation of astragalus-ginkgo biloba-bran are finer, Chinese medicine has a delicate fragrance. The yeasts are large and small, and grow vigorously; the product after the fermentation of hawthorn-ginkgo biloba-bran has small particles, thick, and strong hawthorn flavor, with small yeasts; and the fermentation product particles of ginkgo biloba are relatively small. It is fine, with a wine aroma, and the yeast is full of form. The results of multiple comparisons showed that the number of yeasts produced by the mixed fermentation of Astragalus-Ginkgo biloba in a ratio of 1:4 reached a very significant level with other combinations. Compared with pine needles-ginkgo biloba and hawthorn-ginkgo biloba, the crude protein content in the culture of Astragalus-Ginkgo biloba fermentation reached a very significant level.

[0033] 4. The results of analysis of variance showed that the type of Chinese medicine has a very significant influence on the number of yeasts and the content of crude protein in the yeast culture, while the ratio of Chinese medicine to ginkgo biloba and bran has no significant influence on the two (Table 3). From the above results, it can be concluded that it is more appropriate to use astragalus-ginkgo biloba as the fermentation substrate for fermentation, and the ratio of astragalus-ginkgo biloba-bran is 1:2 to 4:1. From an economic point of view, astragalus can also be considered- The ratio of ginkgo leaf to bran is 1:4:1.

[0034] Table 3 Variance analysis table of different Chinese medicine-ginkgo biloba combined fermentation

[0035]

[0036] Preparation Example 2

[0037] 1. Transfer the Candida Ruanan strain to the slant medium and culture at 28°C for 20 hours. The above slant medium is a PDA medium. The preparation process is as follows: 200g potatoes are boiled in water and 20g sucrose and 10g agar are added. , Mix well, then add water to make the volume up to 1000mL.

[0038] 2. The Candida utilis obtained in step (1) was inoculated into the liquid seed culture medium on a clean bench, and cultured on a shaker (28° C., 200 rpm) for 24 hours. The preparation process of the liquid seed culture medium is as follows: Dissolve 20 g of glucose, 10 g of peptone, and 10 g of yeast extract in water, and then dilute to 1000 mL with water. The liquid medium was sterilized at 121°C for 20 minutes. The content of yeast in the obtained seed culture solution is 9×10 9 Pieces/mL, store in a refrigerator at 4°C, which can be stored for 1 month.

[0039] 3. Dry and grind astragalus, ginkgo biloba, and bran into fine powder, sieve 40 mesh, and mix evenly according to the ratio of astragalus, ginkgo biloba, and bran to 1:4:1 to obtain a mixture of astragalus-ginkgo biloba-bran material.

[0040] 4. Weigh 3g of sucrose, (NH4) in proportion 2 SO 4 4g, KH 2 PO 4 2g, MgSO 4·7H 2 O 0.5g, dissolved in water, constant volume, adjusted with acetic acid to pH 5.5 to 6.0, to obtain 100ml of nutrient solution.

[0041] 5. Add 10.2ml nutrient solution for every 6g astragalus-ginkgo biloba-bran mixture to mix the above two ratios, stir evenly, and sterilize at 121°C for 30min. Among them, the volume of 1g of astragalus-ginkgo biloba-bran mixture is 3mL, and its own water content is about 10%. After adding the nutrient solution, 18ml of solid medium with a water content of 60% (g/mL) is obtained.

[0042] 6. On the aseptic workbench, insert the Candida rudimentum seed solution into the sterilized solid medium at a volume ratio of 7.43% for fermentation, control the temperature at 28°C, humidity at 95%, and cultivate for 60 hours.

[0043] 7. Dry and crush the fermented product at 40°C, which is the Astragalus-Ginkgo biloba biological feed additive.

[0044] Preparation Example 3

[0045] 1. Transfer the Candida Ruanan strain to the slant medium and culture for 24 hours at 25°C. The above slant medium is a PDA medium. The preparation process is as follows: Cut 200g potatoes into small pieces and boil them in water Filter the potato juice with double gauze for 20-30 minutes, add 15g of sucrose and stir to dissolve, then add 20g of agar, continue heating, make the agar completely melt and become transparent, and finally add water to make the volume to 1000mL.

[0046] 2. The Candida utilis obtained in step (1) was inoculated into the liquid seed culture medium on the ultra-clean workbench, and cultured on a shaker (25° C., 200 rpm) for 30 hours. The liquid seed culture medium formula is: 20g glucose, 6g peptone, 10g yeast extract, dissolved in distilled water to a constant volume to 1000mL. The liquid medium was sterilized at 121°C for 20 minutes. The content of yeast in the obtained seed culture solution is 6×10 9 Pieces/mL, put in the refrigerator at 4℃, can be stored for 1 month.

[0047] 3. Dry and grind astragalus, ginkgo biloba, and bran into fine powder, sieve 40 meshes, and mix evenly according to the ratio of astragalus, ginkgo biloba, and bran to 1:2:1 to obtain a mixture of astragalus, ginkgo biloba and bran. material.

[0048] 4. Weigh 3g of sucrose, (NH4) in proportion 2 SO 4· 3.5g, KH 2 PO 4 2g, MgSO 4 ·7H 2 0.5g of O, dissolved in water to a constant volume, adjust the pH to 5.5-6.0 with acetic acid to obtain 100ml of nutrient solution.

[0049] 5. Add 11.1ml nutrient solution for every 6g of astragalus-ginkgo biloba-bran to mix the above two ratios, stir evenly, and sterilize at 121°C for 30min. Among them, the volume of 1 g of astragalus-ginkgo-bran mixture is 3 mL, and its water content is about 10%. After adding the nutrient solution, 18 mL of solid medium with a water content of 65% (g/mL) is obtained.

[0050] 6. In a sterile workbench, insert the Candida ruanana seed solution into a sterilized solid medium at a volume ratio of 10% for fermentation, control the temperature at 30°C, and the humidity at 90%, and cultivate for 48 hours.

[0051] 7. Dry and crush the fermented product at 40°C, which is the Astragalus-Ginkgo biloba biological feed additive.