Lactobacillus acidophilus and application thereof and feed additive thereof and premix compound thereof
A technology of Lactobacillus acidophilus and feed additives, applied in the biological field, can solve problems such as decreased immunity, enhanced drug resistance of bacteria, and imbalance of animal intestinal flora, and achieves strong stress resistance, reduced antibiotic addition, and active The effect of high bacterial content
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Embodiment 1
[0023] Example 1 Screening and identification of Lactobacillus acidophilus CGMCC No.5093
[0024] 1. Primary screening
[0025] Take 5g of fresh pig feces samples, soak them in 20ml of sterile 0.85% normal saline, shake the samples to break up the samples, make the samples fully contact with the normal saline, add the centrifuged bacteria sediment into 10ml of artificial gastric juice, and let it stand at 37°C for 2h , and then centrifuged to collect the bacteria, add the precipitate of the bacteria to 10ml of artificial bile salt, let stand at 37°C for 4 hours, collect the bacteria, dilute and spread on LB medium. After culturing in a 37°C incubator for 12-24 hours, the colonies were purified and isolated. The obtained bacteria were preserved and further re-screened.
[0026] Colony morphology: the colony is round, smooth, opaque, milky white, with smooth edges, Gram-positive bacteria; observed under a microscope after Gram staining, a single bacterium is slender rod-shaped...
Embodiment 2
[0032] Example 2 Experiments on the probiotic function and stress resistance of Lactobacillus acidophilus CGMCC No.5093
[0033] The Lactobacillus acidophilus CGMCC No.5093 obtained by using MRS medium was tested for acid resistance, bile salt resistance and antibacterial activity.
[0034] (1) Acid resistance test of Lactobacillus acidophilus
[0035] Preparation of artificial gastric juice: The preparation method of artificial gastric juice is: 1% pepsin, 0.85% sodium chloride, adjust the pH value to 2.0, 3.0, 4.0 with concentrated hydrochloric acid, filter and sterilize through a bacterial filter for later use; the test results are shown in the table 1.
[0036] Table 1 Lactobacillus acidophilus acid resistance test results
[0037]
[0038] (2) Lactobacillus acidophilus bile resistance liquid test: the preparation method of artificial bile salt is: respectively add 0.1%, 0.2%, 0.3% pig bile salt (analytical pure) in liquid broth, 115 ℃ autoclaving backup used; the te...
Embodiment 3
[0046] The preparation of embodiment 3 Lactobacillus acidophilus CGMCC No.5093 feed additive
[0047] 1) Plate culture rejuvenation: Inoculate Lactobacillus acidophilus strains on the MRS plate medium, culture at 37°C for 24 hours, rejuvenate Lactobacillus acidophilus, and form a single colony, pick a single colony on the inoculation medium, 37 Cultivate for 24 hours at ℃;
[0048] 2) Preparation of primary seeds: transfer the Lactobacillus acidophilus cultured in step (1) into a 1L shaker flask filled with 300ml of MRS liquid medium, and culture at 37°C for 24 hours until the late logarithmic period to obtain primary seeds ;
[0049] 3) Preparation of secondary seeds: transfer the primary seeds prepared in step 2) to a 100L seed tank equipped with 80LMRS seed medium, at a temperature of 37°C, turn 60rpm, stir once every 45 minutes, and each stirring time is 15 minutes, cultivated for 20 hours, and obtained the secondary seed solution.
[0050] 4) Preparation of Lactobacill...
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