Preparation method and application of Bacillus subtilis A16
A technology of Bacillus subtilis and bacteria powder, which is applied in the field of probiotics to achieve the effects of simple and fast preparation process, increasing production performance, and improving animal immunity
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Embodiment 1
[0025] The preparation of embodiment 1 Bacillus subtilis A16
[0026] Take 40mL of the overnight cultured Bacillus subtilis A16 and transfer it into 500ml of fermentation medium (initial inoculum size is 8%), ferment at 30°C for 20 hours, the pH value is 7.0, and the rotation speed is 200rpm. After fermentation, the bacteria were spray-dried, and the bacteria powder was stored at 4°C.
[0027] The composition of the fermentation medium is: sucrose 10g / L, peptone 5g / L, beef extract 5g / L, yeast extract powder 1.5g / L, manganese sulfate 0.0004g / L, pH 7.0.
[0028] Plate colony counting method detects that the number of viable bacteria in the powder is about 10 10 CFU / g.
[0029] The operation method of the plate colony counting method is: after the sample to be tested is diluted by an appropriate multiple, the microorganisms in it are fully dispersed into single cells, and a certain amount of diluted sample is applied to the plate, and after cultivation, each single cell grows a...
Embodiment 2
[0032] The antibacterial effect of the preparation of embodiment 2 bacillus subtilis A16
[0033] Thaw the LB solid medium and cool to 45°C. Add the overnight cultured E.coli K88, K99 and Staphylococcus aureus (1 μL bacterial solution per ml medium), shake and mix, pour into a sterile plate, solidify horizontally, gently place 4 bacteria on each agar plate Oxford cups should be equally spaced. Add 200 μL of the bacteria solution to be tested (Bacillus subtilis A16) into each Oxford cup, cover the dish, carefully move it to a 37°C incubator, and place the dish for static culture. After cultivating for 20 hours, open the lid of the dish, remove the Oxford cup, and measure the diameter of the inhibition zone with a caliper (Table 1). The experimental results showed that Bacillus subtilis A16 had strong antibacterial ability.
[0034] pathogenic bacteria Escherichia coli K88 Escherichia coli K99 Staphylococcus aureus Bacillus subtilis A16 (cm) 1.1 1.2 0.8...
Embodiment 3
[0035] The application of embodiment 3 bacillus subtilis A16 as feed additive
[0036] 90 healthy 28±2-day-old weaned piglets (average body weight 7.6±0.15kg) were randomly divided into 5 groups according to the principle of half male and half female, each group had 3 replicates, and each replicated 6 piglets, group 1 was The blank control group was fed the basal ration, group 2 was fed the basal ration with a chlortetracycline addition of 75 mg / kg, group 3, group 4 and group 5 were fed with 10 9 CFU / kg, 10 10 CFU / kg, 10 11 CFU / kg Basal diet of Bacillus subtilis A16 powder. Food and water were provided ad libitum. During the experiment period, feeding and management were carried out according to the conventional method of the pig farm. On the 28th day of the experiment, feeding was stopped at 8:00 pm, and water was supplied as usual. The pigs were weighed on an empty stomach at 8:00 am the next day, and the average daily gain, average daily feed intake and feed conversion ...
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