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Lactobacillus salivarius capable of inhibiting Candida albicans growth, and separation method thereof
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A technology of Lactobacillus salivarius and Candida albicans, applied in the field of microorganisms
Active Publication Date: 2014-02-05
ZHEJIANG UNIV
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However, in many cases, even if the three treatment methods are applied at the same time, the patient cannot completely get rid of the trouble of Candida albicans
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Embodiment 1
[0031] 1) Take 0.5 g of fresh feces samples from different healthy people and add them to 10 ml of enrichment medium respectively, 37 0 C. Under anaerobic conditions, enrich and cultivate for 24 hours; the ingredients of the enrichment medium are: 5 grams of peptone, 5 grams of beef extract, 2 grams of yeast extract, 8 grams of glucose, 2 grams of lactulose, and 0.5 grams of Tween 80 , 2 grams of dipotassium hydrogen phosphate, 2 grams of sodium acetate trihydrate, 2 grams of diammonium citrate, 0.2 grams of magnesiumsulfate heptahydrate, 0.03 grams of manganesesulfatetetrahydrate, 950 ml of distilled water, pH6.2.
[0032] 2) After diluting the above sample enrichment medium 1000 times, take 100 microliters and spread it on the screening plate, 37 0 C anaerobic culture for 48 hours; the composition of the screening plate medium is: 5 grams of yeast extract, 8 grams of tryptone, 10 grams of glucose, 4 grams of potassium dihydrogen phosphate, 0.02 grams of ferroussulfate, 0...
Embodiment 2
[0037] 1) Take 0.2 g of vaginal secretion samples from different healthy women and add them to 5 ml enrichment medium, 37 0 C, under anaerobic conditions, enrich and cultivate for 24 hours; the ingredients of the enrichment medium are: 8 grams of peptone, 6 grams of beef extract, 4 grams of yeast extract, 6 grams of glucose, 3 grams of lactulose, 0.7 grams of Tween 80, Dipotassium hydrogen phosphate 1.5 g, sodium acetate trihydrate 4 g, diammonium citrate 1 g, magnesium sulfate heptahydrate 0.3 g, manganese sulfate tetrahydrate 0.04 g, distilled water 950 ml, pH 6.2.
[0038] 2) Dilute the above sample enriched culture medium by 5000 times, take 200 microliters and spread it on the screening plate, 37 0 C anaerobic culture for 24 hours; the ingredients of the screening plate medium are: 4 grams of yeast extract, 10 tryptones, 8 grams of glucose, 48 grams of potassium dihydrogen phosphate, 0.04 grams of ferrous sulfate, 0.6 grams of magnesium sulfate heptahydrate, three 25 gra...
Embodiment 3
[0043] 1) Take 200 microliters of saliva samples from different healthy people and add them to 5 milliliters of enrichment medium, 37 0 C. Under anaerobic conditions, enrich culture for 48 hours; the enrichment medium components are: 6 grams of peptone, 5.5 grams of beef extract, 3 grams of yeast extract, 7 grams of glucose, 3 grams of lactulose, 0.7 grams of Tween 80, 1.5 grams of dipotassium hydrogen phosphate, 2.5 grams of sodium acetate trihydrate, 1.5 grams of diammonium citrate, 0.3 grams of magnesium sulfate heptahydrate, 0.04 grams of manganese sulfate tetrahydrate, pH 6.5, 960 ml of distilled water.
[0044] 2) Dilute the above sample enriched culture medium by 4000 times, take 100 microliters and spread it on the screening plate, 37 0 C anaerobic culture for 48 hours; the ingredients of the screening plate medium are: 4 grams of yeast extract, 9 grams of tryptone, 10 grams of glucose, 5 grams of potassium dihydrogen phosphate, 0.05 grams of ferrous sulfate, 0.8 grams...
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Abstract
The present invention discloses a strain of Lactobacillus salivarius capable of inhibiting Candida albicans growth, and a separation method thereof, wherein the Latin name is Lactobacillus salivarius LI01, the Lactobacillus salivarius LI01 is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.7045. The present invention discloses morphological characteristics of the Lactobacillus salivarius LI01, wherein the morphological characteristics comprise rod-shaped bacteria, no spore production, no motility, and positive Gramstaining. The present invention discloses main components of the whole-cellfatty acid of the Lactobacillus salivarius LI01, wherein the main components comprise: about 38.48% of 16:0, about 8.29% of 18:1w9c, about 14.68% of 19:0cyclow10c / 19w6, about 5.94% of 18:0, about 12.75% of 19:0cyclow10c / 19w6, about 11.55% of 19:0cyclow8c, about 12.75% of 19:1w6c, and about 14.68% of 18:1w7c. The invention further discloses the whole sequence of 16S ribosomal DNA (16SrDNA) of the Lactobacillus salivarius LI01. The separation screening method of the present invention comprises three steps: carrying out enrichment culture with a MRS nutrient culture medium, adopting a Candida albicans covering layer to carry out primary screening, and adopting liquid co-culture to carry out re-screening. According to the present invention, the method is rapid and efficient, and the obtained Lactobacillus salivarius LI01 has strong Candida albicans resistance ability.
Description
technical field [0001] The invention belongs to the technical field of microorganisms, and relates to a lactobacillus salivarius capable of inhibiting the growth of Candida albicans and an isolation method thereof. Background technique [0002] Candida albicans, also known as Candida Albicans, is an opportunistic pathogenic fungus that widely exists in nature and human skin, oral cavity, upper respiratory tract, intestinal tract and vagina. In a healthy state, the number of Candida albicans on the surface or body of the human body is small and does not cause disease. When the body's immunity is weakened or the microecology is damaged and other special circumstances, Candida albicans grows in large numbers, causing acute, subacute or chronic infections, and is the most common fungal disease. Factors that may induce Candida albicans infection include: Acquired Immunodeficiency Syndrome (AIDS), genetic defects, use of antibiotics and immunosuppressants, tumors and tumor radiot...
Claims
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