Lactobacillus sp. W8173 and application thereof

A Lactobacillus and germ technology, applied in the field of microorganisms, can solve problems such as disease deterioration, bee product residues, food safety threats, etc., to achieve the effect of environmental protection of feed and improvement of symptoms

Active Publication Date: 2021-08-06
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The occurrence of European larval rot has obvious seasonality, and the bee colony is prone to onset in early spring and autumn. Although the honeybee larvae mainly infect the honeybee larval stage, the adult worker bees will carry the honeybee larvae, causing the European larval rot to spread even in the bee colony. It is transmitted between different bee colonies and bee farms, and the pathogenic mechanism of Melilococcus apiformis is not yet clear
[0004] Usually the bee farms abuse antibiotics because there is no effective treatment method during treatment, which not only leads to the development of drug resistance of pathogenic bacteria, but also destroys the microecological balance of the bee intestinal tract and further aggravates the disease
At the same time, the application of antibiotics will lead to residues in bee products, affecting product quality and posing a threat to food safety
[0005] At present, there is no relevant report on the use of Lactobacillus in the control of European larval rot disease

Method used

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  • Lactobacillus sp. W8173 and application thereof
  • Lactobacillus sp. W8173 and application thereof
  • Lactobacillus sp. W8173 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Isolation and Identification of Lactobacillus W8173

[0021] Apis mellifera was captured from Jilin, and the complete intestine of the bee was dissected out, and the intestinal wall was ground and broken by manual grinding in 100 μL 25% (v / v) glycerin with a grinding rod, and stored in a -80°C ultra-low temperature medical refrigerator; The samples were lined in three zones on the lactic acid bacteria medium (MRS), and cultured in a 35°C, 5% carbon dioxide incubator for 2-3 days. Observe the shape of the colony, pick a single colony from it and re-mark eight sub-areas on the MRS medium, and purify and cultivate it in a 35 ℃, 5% carbon dioxide incubator; select several single colonies on the medium, number them separately, and select Twenty single colonies were subjected to 16S rRNA sequencing.

[0022] Bacterial total DNA was extracted using the Bacterial Genomic DNA Extraction Kit of Tiangen Biochemical Technology Co., Ltd., using 16S rRNA universal primers...

Embodiment 2

[0024] Embodiment 2: the preparation of freeze-dried bacterial agent

[0025] Lactobacillus melius W8173 was activated with MRS medium, the inoculum size was 3%, the activated strain was transferred to MRS liquid medium according to the 4% inoculum size, cultured at 37°C for 25 h, centrifuged (5000 g, 15 min ) to harvest the thallus and set aside. Add the above bacteria into the protective agent (1% trehalose, 0.5% sodium glutamate, 1% skim milk, 0.25% ascorbic acid, 0.25% dextran, g / 100mL distilled water) according to the volume ratio of 3:1, mix well spare. Before freeze-drying, the samples were pre-frozen in -80°C ultra-low temperature refrigerator for 3 hours, followed by three-stage freeze-drying, which was freeze-dried at 512 Pa for 6 hours, then at 256 Pa for 5 hours, and then dried at 103 Pa. Freeze-drying at Pa until a powder product is obtained. Store the freeze-dried bacteria powder at 4 ℃ and 25 ℃ respectively, measure the number of viable bacteria of each bacte...

Embodiment 3

[0027] Embodiment 3: Lactobacillus W8173 antibacterial experiment in vitro

[0028] (1) The configuration of the aseptic supernatant of Lactobacillus W8173 in the test product: inoculate the lyophilized bacterial preparation of Lactobacillus W8173 in the above experiment into the lactic acid bacteria bacterial culture medium (52 ​​g of MRS broth medium powder, add distilled water to 1 L , sterilized at 121 ℃ for 15 min) for 5 days, centrifuged the culture solution through a low-temperature high-speed centrifuge at 8000 g for 10 min, took the supernatant, and finally filtered the supernatant through a 0.22 μm pore size filter membrane to obtain Lactobacillus W8173 without bacterial supernatant;

[0029] (2) The configuration of the concentrated sterile supernatant of Lactobacillus W8173 of the test product: Concentrate 1 L of the sterile supernatant of Lactobacillus W8173 into 100 mL by rotary evaporator (55 ℃) to obtain concentrated Lactobacillus W8173 sterile supernatant;

...

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Abstract

The invention discloses lactobacillus sp. W8173 which is preserved in the China General Microbiological Culture Collection Center with the preservation number of CGMCC NO. 21787. The strain can alleviate the symptom of European foulbrood of bees, and in an in-vitro bacteriostasis experiment, the strain can directly inhibit melissococcus pluton; in the in-vivo experiment, single-bacterium bees fed with lactobacillus sp. W8173 have a relatively good inhibition effect on melissococcus pluton; and by separating supernant of a lactobacillus sp. W8173 culture solution, it is found that a secretory product of lactobacillus sp. W8173 has an inhibitory effect on melissococcus pluton. The preparation of the present invention is environmentally friendly, free of pollution, free of influence on bee product quality and suitable for industrial production and market popularization and application.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, in particular to a lactobacillus ( lactobacillus sp.) W8173 and its application. Background technique [0002] bee( Apis mellifera ) is an important agricultural pollinator, which not only plays an important role in the construction of ecological agriculture and the supply of bee products, but also plays an important role in maintaining biodiversity and ecological balance through pollination of different vegetation. At present, about 36 kinds of crops such as apple, cotton, rapeseed and soybean in my country rely heavily on the pollination of bees, and the annual pollination value reaches 304.2 billion yuan. However, due to the reduction of honey plant diversity and habitat, interspecific competition, infection of pathogenic microorganisms and parasites in recent years, the number of bee populations in the world has been declining year by year. Among them, the infection of pathogenic m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K10/16A23K10/18A23K50/90A61K35/747A61P31/04C12R1/225
CPCA23K10/16A23K10/18A23K50/90A61K35/747A61P31/04
Inventor 郑浩郎浩宇张雪王小斐胡小松
Owner CHINA AGRI UNIV
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