Lactobacillus plantarum HL-16 and application thereof
A technology of Lactobacillus plantarum and HL-16, which is applied in the field of Lactobacillus plantarum HL-16, can solve the problems of poor comprehensive performance of probiotics, decreased growth performance, and increased breeding costs, and achieve good tolerance and strong acid production ability , the effect of promoting animal growth
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preparation example Construction
[0055] (1) Preparation of test bacteria:
[0056] Inoculate the test bacteria (Escherichia coli, Staphylococcus aureus, Salmonella suis, Shigella dysenteriae) stored in a -80°C refrigerator into LB liquid medium (each 1000mL medium contains tryptone 10g, sodium chloride 10g , yeast powder 5g, the balance is water), the reference substance is LB liquid medium without the test bacteria (each 1000mL medium contains tryptone 10g, sodium chloride 10g, yeast powder 5g, the balance is water) , are placed in 38 ℃, 180r / min constant temperature shaker culture. Use a UV spectrophotometer (OD600) to measure the absorbance of the bacterial solution every four hours, so that the bacterial content of the bacterial solution is 10 6 -10 8 CFU / mL;
[0057] (2) Preparation of antibacterial agent:
[0058] Weigh 1.0 g of solid sample, add 10 mL of absolute ethanol, mix well, and filter with filter paper. Draw 1 mL of sample and dilute three gradients of 10 with absolute ethanol -1 、10 -2 ...
Embodiment 1
[0062] Example 1: illustrates how to obtain Lactobacillus plantarum HL-16 by screening.
[0063] (1) Plate primary screening and purification: add 10g of chicken manure from a farm in Luoyang to 90mL sterile water, shake well. After shaking well, take 1mL sample and add it to 9mL sterile water, mix it on a vortex mixer, then take 1mL sample and add it to 9mL sterile water, and so on. Select three appropriate concentration gradients to coat solid culture plates in sequence (each 1000 mL medium contains 10 g peptone, 15 g yeast powder, 20 g glucose, 3 g skimmed milk powder, 1.2 g Vc, 1.8 g cysteine, 5 g beef extract, 5 g CH 3 COONa, 2g triammonium citrate, 2gKH 2 PO 4 , 0.25gMnSO 4 , 0.58gMgSO 4 , 1mL Tween 80, 2% agar powder, 1% calcium carbonate, the balance is water, adjust the pH to about 7), culture at 36°C for 36-48h, select the strain that produces a transparent circle on the plate, and set aside. Purify the initially screened bacterial strains on a solid medium plat...
Embodiment 2
[0067] Example 2: illustrates how to identify Lactobacillus plantarum HL-16.
[0068] (1) Separation and purification of a single colony slant, sent to Sangon Biotech (Shanghai) Co., Ltd. for sequencing, the results are shown in the 16S rDNA sequence table of the strain HL-16, and the MEGA7.0 software was used to construct the strain HL-16 Phylogenetic tree (see image 3 ).
[0069] (2) According to the morphological characteristics, physiological and biochemical characteristics and 16S rDNA sequence of the strain HL-16, it is determined that HL-16 is Lactobacillus plantarum ( Lactiplantibacillus plantarum ).
[0070] (3) Preserve the strain HL-16 identified as Lactobacillus plantarum. The preservation unit is the General Microbiology Center of the China Committee for the Preservation of Microbial Strains, and the preservation place is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. Institute, CGMCC N...
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