Swine lactobacillus crispatus and application thereof
A technology of Lactobacillus crispatus, strains, applied in the field of microbiology
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Embodiment 1
[0035] The isolation of embodiment 1 lactic acid bacteria strain
[0036] Collect the fresh feces of suckling piglets, weigh 25g, place it in a glass beaded triangular flask filled with 225ml of sterile normal saline, shake it well, then draw 1.0ml of bacterial liquid into a flask filled with 9.0ml of sterile normal saline In test tubes, this dilution is 10 -1 , repeat the above process, and make 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 of bacteria liquid. choose 10 -4 、10 -5 、10 -6 For three dilutions, draw 0.1ml of the bacterial solution and drop it on the above-mentioned medium plate, culture in an anaerobic tank at 37°C for 24-48 hours, and pick typical colonies for API50 and 16SrDNA identification. According to the lactic acid bacteria fermenting sugar and producing acid to make the medium turn yellow to determine the sugar utilization of the strain, the colonies that produce yellow circles and different shapes are purified, cultured, Gram stained, microscopic examinati...
Embodiment 2
[0037] The screening test of embodiment 2 lactic acid bacteria strains
[0038] After the strain was activated, it was inoculated in MRS liquid medium, and after 24 hours of cultivation, a certain amount of bacterial liquid was taken to measure the OD600nm value, and the growth performance of the strain was observed with no inoculation as the control. At the same time, the strain was inoculated in MRS liquid medium and cultured for 24 hours, 5ml of fermentation broth was taken and centrifuged at 6000rpm for 5 minutes, added to 5ml of liquid medium containing 0.2% bile salt pH=3, incubated for 2h, and then inoculated with 10% Incubate in medium for 4h, and measure OD with culture without any treatment as control 600nm value, and calculate the acid-resistant and bile-resistant performance of the strain.
[0039] Screened by growth, acid and bile salt resistance tests, see figure 1 and figure 2 , confirm that the strain R2 (in the figure) is the target strain (the laboratory ...
Embodiment 3
[0040] The identification of embodiment 3 Lactobacillus crispatus
[0041] The strain R2 (the laboratory number is L.crispatus ZLC020) was sent to the China Industrial Microorganism Culture Collection Management Center for identification, and the physiological and biochemical characteristics (Table 1) and molecular biological identification confirmed that the strain was Lactobacillus crispatus ( Lactobacillus crispatus). The strain was tested on October 22, 2015 in the General Microbiology Center of China Microbiological Culture Collection Management Committee (Address: No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Zip Code: 100101) Preserved, the strain name is Lactobacillus crispatus (Lactobacillus crispatus) ZLC020, and its preservation number is CGMCC No.11531.
[0042] Table 1 Physiological and biochemical characteristics of pig-derived Lactobacillus crispatus
[0043]
[0044] Note: "+" in the ta...
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