Bacillus and application thereof
A technology of Bacillus coagulans and Bacillus coagulans, applied in the field of microorganisms, can solve the problems that the number of viable bacteria of Clostridium butyricum Bacillus coagulans needs to be increased, the problem of contamination is not easy to solve, and the cooling cost is high, so as to enhance animal immunity and resist The effect of strong reversibility and improved palatability
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Embodiment 1
[0041] The screening of embodiment 1 bacillus coagulans
[0042] 1. The medium (liquid) and its formula (g / L) used for strain screening are as follows:
[0043] Enrichment medium (nutrient broth medium): peptone 10, beef extract 3, sodium chloride 5, pH 7.0,
[0044] Calcium carbonate identification plate medium: glucose 5, peptone 5, yeast extract powder 1, calcium carbonate 0.3, agar powder 10, pH6.8~7.0.
[0045] Lactic acid bacteria proliferation medium (YPG): glucose 5, peptone 5, yeast extract 1, calcium chloride 0.1, pH 6.8-7.0.
[0046] Coagulant spore activation medium (1 / 2YPD medium): glucose 10, yeast extract 5, tryptone 10, pH 6.8-7.0.
[0047] Bacillus coagulans count medium (1 / 2YPD solid medium): glucose 10, yeast extract 5, tryptone 10, distilled water 1000mL, agar powder 15, pH 6.8.
[0048] Bacillus coagulans slant medium: beef extract 5, tryptone 10, sodium chloride 5, pH 7.0±0.1.
[0049] Amylase medium: peptone 10, beef extract 5, sodium chloride 5, sta...
Embodiment 2
[0059] Stress resistance and enzyme production performance research of embodiment 2 bacterial strain
[0060] 1. Simulated gastric juice tolerance study
[0061] Centrifuge the activated candidate strain bacterial liquid at 4000r / min for 10min to collect bacterial precipitates, add the collected bacterial cells to the simulated gastric juice, and culture them statically at 37°C, take samples at 0, 60min, 120min, and 180min to detect the activity Bacteria count and calculate survival rate, the results are shown in Table 2:
[0062] Table 2 Survival rate (%) of different strains after artificial gastric juice resistance treatment
[0063]
[0064] From the results in Table 2, it can be seen that the activity of all strains was lost after being treated with simulated artificial gastric juice, and the survival rate of strain ZC2-1 was the highest after being treated for 120 minutes.
[0065] 2. Research on bile salt tolerance
[0066] Bile salts in the small intestine of ani...
Embodiment 3
[0078] Embodiment 3 strain identification
[0079] In this example, 4 strains ZC2-1, ZA-1, ZB-1 and ZC-9 with good enzyme production, gastric acid resistance, bile salt resistance and heat resistance were identified through morphological and phylogenetic analysis.
[0080] 1. Morphological identification
[0081] The above four strains were activated and inoculated on the YPD solid medium plate, and cultured at 37°C for 24 hours to observe the colony morphology. Then take an appropriate amount of bacterial liquid for methylene blue staining and microscopic examination. The result is as image 3 shown. Depend on image 3 It can be seen that the bacterial strain ZC2-1 forms a white opaque circular colony, and there is a small white protrusion in the middle of the colony ( image 3 a), the thallus is in the shape of a slender rod with a length of 2.5-4.0 μm and a width of 0.4-0.8 μm ( image 3 e). Strains ZA-1 and ZC-9 both formed light milky white round small colonies wit...
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