Lactobacillus paracasei for preventing infant streptococcus infection and application of lactobacillus paracasei
A technology of Lactobacillus and paracheese, applied in medical preparations containing active ingredients, bacteria, antibacterial drugs, etc., can solve the problem of increasing the incidence of necrotizing enteritis, liver and kidney damage and hearing loss in children, and increasing the incidence of hospitalized children. case fatality rate
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Embodiment 1
[0032] Example 1 Screening of Lactobacillus paracasei VHProbi O44
[0033] 1. Bacteria isolation
[0034] Prepare MRS (Man Rogosa Sharpe) broth: pure water 1L, peptone 10g, beef extract 10g, yeast extract 5.0g, sodium acetate 5g, glucose 5g, potassium dihydrogen phosphate 2g, Tween 80 1.0mL, citric acid Diamine 2.0g, calcium carbonate 20g, magnesium sulfate heptahydrate 0.58g, manganese sulfate heptahydrate 0.25g, adjusted pH 6.2-6.5.
[0035] Configure MRS agar medium: add 15g agar to 1LMRS broth.
[0036]According to the 2019 version of the "Human Genetic Resource Bank Ethical Specifications", after signing the project commitment letter and informed consent with the sample provider, according to the standard operating specifications of the biobank, take 1g of fresh fecal samples from healthy infants who have not eaten probiotic preparations within half a year. After being diluted with sterile saline, put it into a sterile sample bag, beat and mix with a homogenizer; take 1...
Embodiment 2
[0046] Embodiment 2, O44 strain identification
[0047] 1. Identification of colony morphology
[0048] The O44 strain was inoculated on the MRS agar medium, and after anaerobic culture at 37°C for 24 hours, the single colony of O44 was milky white, the diameter of the colony was more than 1.0mm, and the surface was smooth. Single or paired, arranged in chains.
[0049] 2. Identification of physiological and biochemical characteristics
[0050] The preparation of the inoculum in this example is as follows: under aseptic conditions, take an appropriate amount of fresh O44 bacterial solution, centrifuge at 5000 rpm / min for 5 min, wash twice with PBS buffer, and then resuspend the bacteria with the same volume of PBS buffer and dilute 50 times as the inoculum.
[0051] 2.1. Salinity tolerance test
[0052] Under sterile conditions, add 190 μL of BSM liquid medium with a salt concentration of 1%, 2%, 3%, 4%, 5%, 6%, 7%, and 8% to a 96-well plate, respectively, for each salt co...
Embodiment 3
[0097] Example 3 Tolerance test of Lactobacillus paracasei VHProbi O44 to artificial gastric juice and artificial intestinal juice
[0098] 1. Preparation of artificial gastric juice
[0099] Weigh 5 g of peptone, 2.5 g of yeast extract, 1 g of glucose and 2 g of NaCl, add 1000 mL of distilled water, adjust pH to 3.0 with dilute hydrochloric acid, and then sterilize at 115 °C for 20 min. Then add 3.2 g of porcine mucosal pepsin before use, shake well to dissolve, and place in a warm water bath for 1 hour in a 37°C water bath shaker to simulate human body temperature.
[0100] 2. Preparation of artificial intestinal juice
[0101] Weigh 5 g of peptone, 2.5 g of yeast extract, 1 g of glucose, KH 2 PO 4 6.8g and 3.0g of bovine bile salt, add 77mL of 0.2mol / L NaOH solution, dilute to 1000mL, adjust pH to 6.8±0.1 with dilute hydrochloric acid or sodium hydroxide solution, and sterilize at 115℃ for 20min. Then add 1g of pancreatin before use, shake well to dissolve, and put it ...
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