Enterococcus faecalis and its application
A technology of Enterococcus faecalis and cephalosporin, applied to Enterococcus faecalis and its application field, can solve the problem of few types of Enterococcus faecalis and the like
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Embodiment 1
[0035] Example 1 Separation, purification and identification of Enterococcus faecalis
[0036] Pick 5-10 g of the collected fresh feces from children, put them in a glycerol tube and transfer them to a refrigerator with a biological ice pack for later use. In a sterile operating table, 1 g of the anaerobic preserved fecal sample of children was transferred to 9 mL of sterile normal saline and made uniform, and then serially diluted 10 times to 1 × 10 -8 , pipette 1×10 -6 , 1×10 -7 , 1×10 -8 100 μL each of 3 serial dilutions in solid medium MRS-CaCO 3 , 3 plates per dilution. After 48 hours of anaerobic culture at a constant temperature of 37 °C, the colonies with the characteristics of lactic acid bacteria on the surface were picked and inoculated into EC liquid medium for expanded culture, and then Gram staining experiments, catalase experiments, and 16S rRNA sequencing were carried out, and it was determined to be fecal Enterococcus. Among them, MRS-CaCO 3 Peptone 10...
Embodiment 2
[0055] Example 2 Study on acid resistance and bile salt resistance of strains
[0056] 2.1 Cultivation of strains to be tested
[0057] EC medium is used for the cultivation of Enterococcus faecalis strains and commercially available Enterococcus faecalis strains. Accurately weigh according to the formula, add distilled water, stir and mix, adjust the pH to 7.4, and sterilize at 121°C for 20min. 2% of the inoculum was used to inoculate Enterococcus faecalis into sterilized EC medium, and aerobic culture was performed at 180rpm and 37°C.
[0058] 2.2 Preparation method of artificial gastric juice and intestinal juice
[0059] Artificial gastric juice: take 16.4 mL of dilute hydrochloric acid, add about 800 mL of water and 10 g of pepsin, shake well, and add water to dilute to 1000 mL.
[0060] Artificial intestinal juice, namely phosphate buffer (containing pancreatin, pH 6.8): take 6.8 g of potassium dihydrogen phosphate, add 500 mL of water to dissolve, adjust the pH to 6....
Embodiment 3
[0095] Example 3 Safety study of Enterococcus faecalis strains
[0096] 3.1 Drug resistance analysis
[0097] Using the drug sensitive paper diffusion method. Dilute freshly cultured Enterococcus faecalis to 10 with normal saline 7 cfu / ml, evenly spread on the M17 petri dish, placed at room temperature for 3-5min, and then placed the drug sensitive paper, the distance between the centers of the paper is not less than 24mm, and the edge of the paper is not less than 15mm from the edge of the agar. Inverted culture in a 37 ℃ incubator, 24h to measure the diameter of the inhibition ring.
[0098] The drug resistance of Enterococcus faecalis and its 30th generation strains was determined by the disc diffusion method, and the results are shown in Tables 5 and 6 below.
[0099] Table 5 Resistance results of Enterococcus faecalis
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[0102] Note: R=resistance; I=intermediate; S=susceptible; -: no zone of inhibition;
[0103] Table 6 Results of drug resistan...
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