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A strain of Lactobacillus reuteri and its application in bee breeding

A technology of Lactobacillus reuteri and bees, which is applied in the field of application of Lactobacillus reuteri LP4 strain in the process of bee breeding, can solve the problems affecting the health of consumers, the imbalance of intestinal flora microecology, and the reduction of the quality of bee products, etc. question

Active Publication Date: 2021-04-20
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to control bee infectious diseases, antibiotics are still mainly used for prevention and treatment, and there are phenomena such as blind drug use and excessive use, which not only lead to the imbalance of intestinal flora microecology, enhance the drug resistance of intestinal microorganisms, and make general drugs ineffective, Moreover, antibiotics will remain in bee products, reducing the quality of bee products, affecting the health of consumers, and reducing the economic benefits of bee farms

Method used

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  • A strain of Lactobacillus reuteri and its application in bee breeding
  • A strain of Lactobacillus reuteri and its application in bee breeding
  • A strain of Lactobacillus reuteri and its application in bee breeding

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Experimental program
Comparison scheme
Effect test

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%, a...

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Abstract

The invention belongs to the field of biotechnology and provides a strain of Lactobacillus reuteri and its application in the bee breeding process. Lactobacillus reuteri was preserved on July 2, 2018 in the General Microbiology Center of China Microbiological Culture Collection Management Committee, the preservation number is CGMCC No.16043, and the classification is named Lactobacillus reuteri; the Lactobacillus reuteri After cultured on MRS agar medium, the colony was round, milky white, with neat edges, smooth surface and raised, opaque. The Lactobacillus reuteri LP4 was independently isolated from the intestines of worker bees of Apis mellifera Apis mellifera, which were raised by beekeepers in Luoping County, Yunnan Province, and freely fed on rapeseed nectar in spring. It has good antibacterial activity and is sensitive to various antibiotics. Press Lactobacillus reuteri LP4 by 10 8 The amount of CFU / mL added to the sugar water fed to bees can improve the micro-ecological structure of the flora in the bee intestines, enhance the intestinal immunity of bees, and increase the survival rate of bees.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a strain of Lactobacillus reuteri and its application in the bee breeding process, in particular to the application of a Lactobacillus reuteri LP4 strain in the bee breeding process. Background technique [0002] Honey bee is a common beneficial insect, belonging to Hymenoptera, Thinwaist suborder, Pintail, Apioidea, Apiaceae insects. Its greatest contribution to human beings is that it can pollinate plants and improve the quality of plant fruits and seeds. At the same time, the honey produced, collected pollen, secreted bee venom and propolis have important applications in medicine, agriculture and industry. As a highly social insect, honeybee's infectious diseases are often explosive and mixed, characterized by fast epidemic speed, wide disease area, and high lethality, which poses a great threat to bee colonies. In order to control bee infectious diseases, antibiotics are still mai...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K50/90A23K10/18A23K20/163C12R1/225
CPCA23K10/18A23K20/163A23K50/90C12R2001/225C12N1/205A23V2400/173
Inventor 林秋叶汪思凡曹振辉郭文清潘洪彬雷清芝钱颖艳黄丝艳叶朋飞余行罗程
Owner YUNNAN AGRICULTURAL UNIVERSITY
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