Lactobacillus reuteri and application thereof in bee breeding process
A Lactobacillus reuteri and honeybee technology is applied in the application field of Lactobacillus reuteri LP4 strain in the bee breeding process, which can solve the problem of enhancing the resistance of intestinal microorganisms, the imbalance of intestinal flora microecology, and reducing the maintenance of nutrients. Bee farm economic benefits and other issues, to achieve the effect of enhancing the intestinal immunity of bees, reducing the application and improving the survival rate of bees
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Embodiment 1
[0024] Screening and identification of Lactobacillus reuteri LP4
[0025] (1) Screening of Lactobacillus reuteri LP4
[0026] (1) Collection of samples
[0027] Put the healthy adult worker bees of Apis mellifera randomly picked from Luoping County, Yunnan Province into a 50mL sterilized centrifuge tube, insert carbon dioxide until the bees are comatose, fix the bees in the ultra-clean workbench, use tweezers to take out the intestines of the bees and dry them. Transfer it into a 1.5mL sterilized centrifuge tube and store it in a 4°C refrigerator for later use. All the equipment used in the experiment was sterilized with ultraviolet light and 75% ethanol.
[0028] (2) Screening of lactic acid bacteria
[0029] Under sterile conditions, the gut samples of healthy adult worker bees of Apis mellifera were taken, added to 10 mL of PBS, homogenized for 5 min, and inoculated into MRS broth medium containing 0.1% vitamin C at a ratio of 1%, 37 ℃ anaerobic culture for 18h. The enr...
Embodiment 2
[0038] Prebiotic properties of Lactobacillus reuteri LP4
[0039] (1) Antibacterial properties of Lactobacillus reuteri LP4
[0040] The inhibitory effect of Lactobacillus reuteri LP4 on the growth and reproduction of major intestinal pathogenic bacteria was studied by agar punching diffusion method. Lactobacillus reuteri LP4 was inoculated in MRS broth medium, anaerobically cultured at 37°C for 24 hours, and the number of bacteria was adjusted to 10 with MRS broth medium. 8 CFU / mL; 4°C, 5000rpm, centrifuge for 10min, collect the supernatant; filter the supernatant through a 0.22μm microporous membrane, adjust the pH to 6.2 with sodium hydroxide; incubate 20μL for 24h with a concentration of 1×10 8 CFU / mL indicator bacteria solution (Escherichia coli, Salmonella typhimurium, Shigella flexneri) was added to the LB solid medium sterilized by high temperature and high pressure and cooled to about 45°C, mixed vigorously and quickly poured into a plate In the medium, after the me...
Embodiment 3
[0049] Effects of Lactobacillus reuteri LP4 on growth and intestinal health of adult worker bees of Apis mellifera
[0050] (1) Effect of Lactobacillus reuteri LP4 on the mortality of adult worker bees of Apis mellifera
[0051] 300 healthy adult worker bees of Apis mellifera were captured from the apiary of Yunnan Agricultural University and randomly divided into two groups. 8 CFU / mL sugar water of Lactobacillus reuteri LP4; the control group and the experimental group were reared in two self-made incubators respectively, and sufficient sugar and water supply were ensured during the experiment; On the 9th day, the bee mortality in the control group and the experimental group was counted respectively. Experimental results such as figure 1 As shown, with the increase of feeding age, the mortality rate of bees in the experimental group and the control group gradually increased; but on the 3rd, 6th, and 9th day, the mortality rates of the experimental group were 15.4%, 26.4%, ...
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