Preparation method of forage bacillus subtilis powder

Abstract

The invention discloses a preparation method of a forage bacillus subtilis powder. The preparation method comprises the following steps: first, the bacillus subtilis strain is inoculated on a slant culture medium to carry on slant culture, then the slant is flushed with a sterile physiological-saline, and a seed bacteria suspension is obtained; second, the seed-bacteria suspension is inoculated in a seed tank with the inoculation amount of 3% to 5%,by volume, to carry on fermentation culture, and a seed bacteria liquid is obtained; third, the seed bacteria liquid is inoculated into a fermentation cylinder to carry on the fermentation culture using a pressure difference method, and a fermentation broth is obtained; and fourth, bacteria sludge is obtained through bacteria-liquid separation of the fermentation broth, and the forage bacillus subtilis powder is obtained through spray drying of the bacteria sludge. Through the exploring of culture medium formulas and fermentation conditions, the problems that the bacteria powder prepared by conventional methods has low viable bacteria count and low spore transformation rate are well solved, the production efficiency is increased, and the cost of enterprises is reduced. The forage bacillus subtilis powder prepared through the method of the present invention has high viable bacteria count and high spore transformation rate.

Description

technical field

[0001] The invention belongs to the technical field of preparation of feed bacterial powder, and in particular relates to a preparation method of feed bacterial powder of Bacillus subtilis. Background technique

[0002] Bacillus subtilis, a kind of Bacillus, aerobic bacteria, single cell 0.7~0.8×2~3μm, uniform coloring, no capsule, perinatal flagella, able to move, Gram-positive bacteria, spore 0.6~0.9 ×1.0~1.5μm, elliptical or columnar, located in the center of the fungus or slightly off, and the fungus does not expand after spore formation. Bacillus subtilis is widely distributed in soil and decayed organic matter, and it is easy to reproduce in subtilis juice. It can use protein, various sugars and starch to decompose tryptophan to form indole. The application of Bacillus subtilis in feed can effectively regulate the balance of gastrointestinal flora in livestock and poultry, inhibit the reproduction of harmful microorganisms; improve the antibody level a...

Examples

Embodiment 1

[0021] Step 1, inoculate the Bacillus subtilis strains on the slant medium for slant culture, the culture temperature is 37°C, and the culture time is 28h; then rinse the slant with sterile physiological saline to obtain a seed suspension; the slant medium The pH value of the medium is 7.0, and the medium is composed of: beef extract 5.0g / L, glucose 7.0g / L, sodium chloride 5.0g / L, peptone 8.0g / L, agar 20.0g / L, and the balance is water; The thalline concentration of the seed bacteria suspension is 5.0×10 8 CFU / mL;

[0022] Step 2: inoculate the seed bacteria suspension described in step 1 in a seed tank with an inoculation amount of 4% by volume to carry out fermentation and culture to obtain a seed bacteria liquid; the conditions for the fermentation and culture of the seed tank are: culture temperature 34°C, The tank pressure is 0.05MPa, the pH value is controlled at 7.2, the ventilation ratio is 1:1.2 (V / V min), the stirring speed is 180r / min, and the fermentation time is 1...

Embodiment 2

[0027] Step 1, inoculate the Bacillus subtilis strain on the slant medium for slant culture, the culture temperature is 37°C, and the culture time is 22h; then rinse the slant with sterile physiological saline to obtain a seed suspension; the slant medium The pH value of the medium is 6.8, and the medium is composed of: beef extract 4.0g / L, glucose 8.0g / L, sodium chloride 6.0g / L, peptone 10.0g / L, agar 18.0g / L, and the balance is water; The thalline concentration of the seed bacteria suspension is 5.0×10 7 CFU / mL;

[0028] Step 2: inoculate the seed bacteria suspension described in step 1 in a seed tank with an inoculation amount of 5% by volume to carry out fermentation and culture to obtain a seed bacteria liquid; the conditions for the fermentation and culture of the seed tank are: culture temperature 35°C, The tank pressure is 0.06MPa, the pH value is controlled at 7.1, the ventilation ratio is 1:1.2 (V / V min), the stirring speed is 220r / min, and the fermentation time is 1...

Embodiment 3

[0033] Step 1, inoculate the Bacillus subtilis strains on the slant medium for slant culture, the culture temperature is 37°C, and the culture time is 30h; then rinse the slant with sterile physiological saline to obtain a seed suspension; the slant medium The pH value of the medium is 7.2, and the medium is composed of: beef extract 3.0g / L, glucose 9.0g / L, sodium chloride 4.0g / L, peptone 9.0g / L, agar 15.0g / L, and the balance is water; The thalline concentration of the seed bacteria suspension is 6.0×10 8 CFU / mL;

[0034] Step 2: inoculate the seed bacteria suspension described in step 1 in a seed tank with an inoculation amount of 3% by volume to carry out fermentation and culture to obtain a seed bacteria liquid; the conditions for the fermentation and culture of the seed tank are: culture temperature 37°C, The tank pressure is 0.08MPa, the pH value is controlled at 7.0, the ventilation ratio is 1:1.2 (V / V min), the stirring speed is 250r / min, and the fermentation time is 8...

technical field

[0001] The invention belongs to the technical field of preparation of feed bacterial powder, and in particular relates to a preparation method of feed bacterial powder of Bacillus subtilis. Background technique

[0002] Bacillus subtilis, a kind of Bacillus, aerobic bacteria, single cell 0.7~0.8×2~3μm, uniform coloring, no capsule, perinatal flagella, able to move, Gram-positive bacteria, spore 0.6~0.9 ×1.0~1.5μm, elliptical or columnar, located in the center of the fungus or slightly off, and the fungus does not expand after spore formation. Bacillus subtilis is widely distributed in soil and decayed organic matter, and it is easy to reproduce in subtilis juice. It can use protein, various sugars and starch to decompose tryptophan to form indole. The application of Bacillus subtilis in feed can effectively regulate the balance of gastrointestinal flora in livestock and poultry, inhibit the reproduction of harmful microorganisms; improve the antibody level a...

Examples

Embodiment 1

[0021] Step 1, inoculate the Bacillus subtilis strains on the slant medium for slant culture, the culture temperature is 37°C, and the culture time is 28h; then rinse the slant with sterile physiological saline to obtain a seed suspension; the slant medium The pH value of the medium is 7.0, and the medium is composed of: beef extract 5.0g / L, glucose 7.0g / L, sodium chloride 5.0g / L, peptone 8.0g / L, agar 20.0g / L, and the balance is water; The thalline concentration of the seed bacteria suspension is 5.0×10 8 CFU / mL;

[0022] Step 2: inoculate the seed bacteria suspension described in step 1 in a seed tank with an inoculation amount of 4% by volume to carry out fermentation and culture to obtain a seed bacteria liquid; the conditions for the fermentation and culture of the seed tank are: culture temperature 34°C, The tank pressure is 0.05MPa, the pH value is controlled at 7.2, the ventilation ratio is 1:1.2 (V / V min), the stirring speed is 180r / min, and the fermentation time is 1...

Embodiment 2

[0027] Step 1, inoculate the Bacillus subtilis strain on the slant medium for slant culture, the culture temperature is 37°C, and the culture time is 22h; then rinse the slant with sterile physiological saline to obtain a seed suspension; the slant medium The pH value of the medium is 6.8, and the medium is composed of: beef extract 4.0g / L, glucose 8.0g / L, sodium chloride 6.0g / L, peptone 10.0g / L, agar 18.0g / L, and the balance is water; The thalline concentration of the seed bacteria suspension is 5.0×10 7 CFU / mL;

[0028] Step 2: inoculate the seed bacteria suspension described in step 1 in a seed tank with an inoculation amount of 5% by volume to carry out fermentation and culture to obtain a seed bacteria liquid; the conditions for the fermentation and culture of the seed tank are: culture temperature 35°C, The tank pressure is 0.06MPa, the pH value is controlled at 7.1, the ventilation ratio is 1:1.2 (V / V min), the stirring speed is 220r / min, and the fermentation time is 1...

Embodiment 3

[0033] Step 1, inoculate the Bacillus subtilis strains on the slant medium for slant culture, the culture temperature is 37°C, and the culture time is 30h; then rinse the slant with sterile physiological saline to obtain a seed suspension; the slant medium The pH value of the medium is 7.2, and the medium is composed of: beef extract 3.0g / L, glucose 9.0g / L, sodium chloride 4.0g / L, peptone 9.0g / L, agar 15.0g / L, and the balance is water; The thalline concentration of the seed bacteria suspension is 6.0×10 8 CFU / mL;

[0034] Step 2: inoculate the seed bacteria suspension described in step 1 in a seed tank with an inoculation amount of 3% by volume to carry out fermentation and culture to obtain a seed bacteria liquid; the conditions for the fermentation and culture of the seed tank are: culture temperature 37°C, The tank pressure is 0.08MPa, the pH value is controlled at 7.0, the ventilation ratio is 1:1.2 (V / V min), the stirring speed is 250r / min, and the fermentation time is 8...