Lactobacillus johnsonii, and preparation and application thereof
A technology of Lactobacillus johnsonii and freeze-dried preparations is applied in the field of microorganisms to achieve the effects of improving body immunity, improving intestinal health and reducing the number of harmful bacteria
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Embodiment 1
[0039] Example 1 Isolation, screening and identification of porcine Lactobacillus johnsonii ZLJ010
[0040] From a standardized breeding pig farm in Beijing, fresh feces of healthy late-pregnancy and lactating sows were collected, put into sterile bags, and immediately brought back to the laboratory in a refrigerated box. Weigh 1 g of feces and add it to 99 mL of sterile saline, and mix well. Take 1mL of the suspension and dilute to 10 with sterile normal saline -5 、10 -6 、10 -7 , 0.1 mL each was spread on MRS solid medium, SL solid medium and LBS solid medium, and cultured anaerobically at 37°C. After 48 hours, the milky white colonies suspected of Lactobacillus were picked for anaerobic pure culture. After 48 h, half of the cultured colonies were picked for H 2 o 2 Enzyme test and Gram stain microscopic examination, all Gram stain positive, H 2 o 2 The strains with negative enzyme test and rod-shaped bacteria can be preliminarily identified as Lactobacillus. Then th...
Embodiment 2
[0055] Embodiment 2 measures the biological characteristics of Lactobacillus johnsonii ZLJ010
[0056] 2.1 Growth curve
[0057] Using OD 600nm The growth characteristics of Lactobacillus johnsonii ZLJ010 were determined based on the index and pH value. see results figure 2 Lactobacillus johnsoni ZLJ010 culture process OD 600nm Curve, image 3 The pH change curve of Lactobacillus johnsonii ZLJ010 culture process.
[0058] Depend on figure 2 , image 3 It can be seen that the OD of Lactobacillus johnsonii ZLJ010 was 600nm The pH value increased rapidly, and the pH value decreased rapidly; during 12~16h, the OD of the bacterial solution 600nm and the change of pH value decreased gradually, indicating that the logarithmic growth phase had been reached; OD after 16h 600nm and pH value did not change significantly, indicating that the bacteria had entered a stable growth period.
[0059] 2.2 Acid resistance test
[0060] 2.2.1 Preparation of artificial gastric juice: 0...
Embodiment 3
[0075] Embodiment 3 prepares Lactobacillus johnsonii preparation
[0076] The lactic acid bacteria preparation is added to the pig’s diet in the form of freeze-dried powder, which is effective and convenient. One is because the freeze-dried lactic acid bacteria preparation is easy to store and has stable activity; the other is because the solid preparation is easy to add to the feed, which is good for mixing uniform. In the production process of freeze-dried preparations, the formula of culture medium, freeze-drying protective agent and freeze-drying process are three important technical links, which directly affect the survival rate of viable bacteria, the number of viable bacteria in freeze-dried preparations and the unit production cost in the production process. . The present invention has carried out technological improvement to these three technical links, has obtained remarkable effect.
[0077] 3.1 Improving the culture medium of Lactobacillus johnsonii ZLJ010
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