Application of bacteroides fragilis in preparation of composition for treating bacterial vaginitis
A technology of Bacteroides fragilis and composition, applied in application, food preparation, antibacterial drugs, etc.
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Embodiment 1
[0017] Example 1 Preparation of Bacteroides fragilis CGMCC No.7280 bacterial liquid
[0018] (1) Isolation of Bacteroides fragilis CGMCC No.7280 from infant feces
[0019] Weigh 0.5 g of stool according to the "normal flora test method", put it in a test tube filled with 4.5 ml of normal saline, and serially dilute it to 10 -1 ~10 -6 , from high concentration to low concentration on the Bd medium plate, placed in an anaerobic gas box and anaerobic gas bag for anaerobic culture. Store at 37°C, open the can for inspection after 2-7 days. A single colony was selected for pure culture, staining microscopy, culture characteristics, and physiological and biochemical testing.
[0020] (2) Bacteroides fragilis CGMCC No.7280 enrichment culture and bacterial liquid preparation
[0021] Select the purified single colony and inoculate it in the improved tryptone broth for enrichment culture. The obtained bacterial solution is centrifuged and precipitated at a speed of 3000r / min for ...
Embodiment 2
[0022] Example 2 Toxicity test of Bacteroides fragilis CGMCC No.7280
[0023] The Bacteroides fragilis bacterium liquid that adopts embodiment 1 preparation carries out following experiment:
[0024] (1) Acute toxicity test in mice
[0025] Sixty C57BR / C mice weighing 22±2 g (half male and half male) were divided into high-dose group, low-dose group and control group, 20 in each group. Each mouse in the high-dose group was intragastrically administered 10 12 cfu / ml bacterial solution 0.5ml, the low dose group was administered orally 10 times a day 9 cfu / ml bacterial solution 0.5ml, and the control group was fed with 0.5ml normal saline every day. Continuous observation for 14 days.
[0026] Experimental results: The mice grew normally, no death was observed, and the LD50 was undetectable.
[0027] (2) Rat toxicity test
[0028] Take 30 SD rats (half male and half female) of 150±20 g, and divide them into 3 groups: high and low dose group and control group. Each rat ...
Embodiment 3
[0033] Example 3 The therapeutic effect of Bacteroides fragilis CGMCC No.7280 on bacterial vaginosis
[0034] Animal model establishment: Escherichia coli (ATCC25922) and Staphylococcus aureus (ATCC25923) strains were recovered and passaged, and prepared with sterile saline to a concentration of 1.8*10 9 / ml of the bacterial solution, mixed in a ratio of 1:1 to form an infectious bacterial solution. First wash the mouse vagina with sterile PBS solution (pH8.5) for 3 times (every interval of 5 minutes), then apply sterile paraffin oil to the No. 5 scalp needle silicone tube, and slowly insert it into the mouse vagina for about 1 cm~1. cm, the injection volume of each bacteria is 0.025ml / 100g, 1 time / day. After the inoculation of bacteria, the vaginal lesions of the mice were observed daily, and vaginal secretions were stained for microscopic examination. When the mouse vagina was obviously congested, red and swollen, accompanied by a large amount of purulent secretions, a ...
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