A kind of Lactobacillus acidophilus la88 for improving vaginal environment and its culture method and application
A technology of Lactobacillus acidophilus and Lactobacillus gasseri, applied in the field of Lactobacillus acidophilus LA88 and its culture, can solve problems affecting the number of inherent probiotics in the vagina of women, bacterial vaginosis, vaginal microecological flora imbalance, etc. To solve the problem of vaginal bacterial infection, relieve inflammation and inhibit pathogenic bacteria
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Embodiment 1
[0042] The present embodiment provides a kind of Lactobacillus acidophilus LA88 that improves vaginal environment, and its separation and screening method is as follows:
[0043] The vaginal secretion samples collected from healthy women were selected, diluted 10 times with normal saline with a mass concentration of 0.9%, diluted 3 times, coated on MRS solid medium, and cultured at 37 °C for 48 h. Different forms were picked out. The 3 strains were streaked and purified on the surface of MRS solid medium, and a single colony was picked and expanded at 37°C in MRS liquid medium, and then preserved with 40% glycerol. The in vitro physiological characteristics of the three preserved single bacteria derived from human vaginal secretions were tested, and a single strain with the best growth ability, acid resistance and bile salt ability (artificial simulation), vaginal epithelial cell adhesion ability and bacteriostatic ability was screened out. strains.
Embodiment 2
[0045] The present embodiment carries out 16S rRNA molecular biology identification to Lactobacillus acidophilus LA88 screened in Example 1, and the steps are as follows:
[0046] The strains stored at -80°C were taken out, inoculated in a centrifuge tube containing 30 mL of MRS liquid medium at a proportion of 2%, cultured at 37°C for 20 h, and then centrifuged at 10,000 rpm for 5 min, and the supernatant was removed. liquid to collect bacteria. The genome of the strain was extracted, and general bacterial primers were added for PCR amplification, and the amplified product was sent to Shanghai Sangon Bioengineering Co., Ltd. for sequencing and identification. The strain was sequenced and analyzed, and its 16S rDNA sequence was shown in SEQ ID No: 1. The sequences obtained by sequencing were compared in GeneBank, and the results showed that the strain was indeed Lactobacillus acidophilus.
[0047] SEQ ID No: 1:
[0048] CGCGTATCTGACTGCTTAGACGGCTCCTTCCCGAAGGTTAGGCCACCGGCTTTG...
Embodiment 3
[0050] The present embodiment optimizes the culture conditions of Lactobacillus acidophilus LA88, and the steps are as follows:
[0051] Lactobacillus acidophilus LA88 was inoculated into MRS liquid medium, and cultured at different temperatures of 10-50 °C for 48 hours. During the cultivation process, the OD of the culture solution was measured by a microplate reader at intervals. 600 numerical value. The results showed that Lactobacillus acidophilus LA88 at 35-42°C, pH at 6.0 The best growth is under 0.2, and the stable growth period can be reached after culturing for 20-25 h.
[0052] According to the API bacterial identification standard, use API 50CHL medium (basal medium, consisting of API 50CH test strips of 48 fermentable carbohydrates) and API 50CH test strips to carry out sugar fermentation reaction on Lactobacillus acidophilus LA88 to detect its carbon source Use capacity. The principle of the method is: use the strain to be tested to make a suspension and inocu...
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