Lactobacillus salivarius and application thereof
A technology of Lactobacillus salivarius, which is applied in the direction of Lactobacillus, application, bacteria, etc., can solve problems such as human health and environmental hazards, and achieve the effects of high cure rate, high adhesion, and good surface properties
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Embodiment 1
[0020] Isolation and purification of embodiment one bacterial strain
[0021] 1.1 Sample collection
[0022] Collect 120 parts of fresh feces from healthy pigs, cattle, and sheep in several farms in Pingliang City, Gansu Province, store them in 50 mL sterile centrifuge tubes, add ice packs, and transport them to the laboratory at low temperature.
[0023] 1.2 Isolation and purification of strains
[0024] Weigh 10 g of feces sample, add it to 90 mL sterile PBS (phosphate buffered saline), mix well and filter, and dilute the filtrate to 10 -4 、10 -5 、10 -6 After doubling, 0.1mL was spread on the MRS plate containing calcium, and cultured anaerobically at 37°C for 24 hours; the suspected colonies of lactic acid bacteria that produced calcium-dissolving circles on the plate were picked and stained with Gram, and the Gram-positive bacteria were picked. A single colony of a rod-shaped strain was streaked and purified on the MRS solid plate, and numbered accordingly; a total of ...
Embodiment 2
[0029] Example 2 Screening and Identification of Probiotic Candidate Beads
[0030] 2.1 Screening of highly stress-resistant lactic acid bacteria
[0031] Determination of acid resistance ability: Inoculate 1 mL of the fresh culture solution of the above 1126 strains of newly isolated lactic acid bacteria into sterile PBS with pH = 2.5. -4 、10 -5 , draw 0.1mL of the diluted solution and spread it on the MRS plate, count and measure the number of viable bacteria after anaerobic culture at 37°C for 24 hours, and calculate the survival rate with the number of viable bacteria at 0h as the control.
[0032] Determination of tolerance to bile salts: Inoculate 1 mL of the fresh culture solution of the above-mentioned 1126 isolates into sterile PBS containing 0.3% bile salts, count and determine the number of viable bacteria after enduring at 37°C for 3 hours, and count the number of viable bacteria as 0h The number was used as the control to calculate the survival rate.
[0033] T...
Embodiment 3
[0053] The biological characteristic research of embodiment Lactobacillus trisalivarius ZLp4b
[0054] 3.1 Acid tolerance study of Lactobacillus salivarius ZLp4b
[0055] The fresh ZLp4b bacterial solution was inoculated in sterile PBS with different pH values (pH gradient 2.0, 2.5, 3.0, 3.5, 4.0) with 5% inoculum, and placed at 37°C for static culture for 0, 2, and 4 hours. Each treatment has 3 replicates. Take 1mL bacterial solution in 9mL PBS, mix and serially dilute to make 10 -4 、10 -5 、10 -6 For the dilution solution, take 0.1 mL of the dilution solution and spread it on the MRS solid plate for counting, and record the number of colonies on the plate after incubating at 37°C for 24 hours. The results of the tolerance of strain ZLp4b to acidic environment are shown in Table 2. ZLp4b can grow under different pH conditions. When the pH is 3.0 or above, the number of viable bacteria after 2h and 4h of cultivation is higher than 1.0×10 8 CFU / mL and basically unchanged,...
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