A strain of Enterobacter hallii and its application in the preparation of antibacterial drugs
A technology of Enterobacter hallii and antibacterial drugs, applied in the field of aquaculture, to improve water quality, prevent hazards, and protect human health and safety
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Embodiment 1
[0013] Example 1: Screening and Identification of Enterobacterhormaechei No. 5
[0014] (1) Search and screening of antagonistic bacteria of Vibrio alginolyticus
[0015] (A) Raw material
[0016] Vibrio alginolyticus
[0017] Antagonistic strains to be screened: 311 strains were isolated from Xisha waters, lobster intestines, sea cucumber intestines, etc.
[0018] Medium reagent used: LB medium, purchased from a reagent company.
[0019] LB medium formula: tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L.
[0020] (B) Preparation of Vibrio alginolyticus solution: Vibrio alginolyticus was inoculated into LB medium with an inoculation loop through aseptic operation, and cultivated overnight at 30°C and 200rpm.
[0021] (C) Isolation and screening of antagonistic bacteria of Vibrio alginolyticus
[0022] (1) Bacteria isolation
[0023] Through aseptic operation, the lobster intestines sampled in Xisha were ground with PBS and diluted into a 10-fold gradient, 100ul...
Embodiment 2
[0034] Embodiment 2: Preparation of antagonistic bacteria preparation (fermentation agent)
[0035] According to the biological characteristics of the best Vibrio alginolyticus antagonistic bacteria Enterobacter halleri No. 5 obtained through screening, a suitable medium was screened, and the antagonistic bacteria was prepared into a fermentation broth by adopting appropriate fermentation conditions.
[0036] Concretely adopted fermentation conditions are as follows:
[0037] Fermentation strain: Vibrio alginolyticus antagonistic bacteria Enterobacter hallii No. 5.
[0038] Fermentation medium formula: tryptone 10g, yeast extract 5g, sodium chloride 10g, distilled water 1L.
[0039] Fermentation temperature: 30°C
[0040] Fermentation cycle: 48-72h
[0041] Rotational speed of fermentation shaker: 150-200rpm
[0042] According to the above fermentation conditions, the Vibrio alginolyticus antagonistic bacteria starter developed by fermentation has a pH of 7.0-7.5. Enterob...
Embodiment 3
[0043] Embodiment 3: Artemia application simulation experiment
[0044] Weigh 0.2g of Artemia (brine shrimp) eggs into 500ml of sterile seawater, and hatch overnight at 30°C under the condition of pumping air at a constant temperature. Use a 12-well plate to do the experiment, prepare in advance the Vibrio alginolyticus antagonistic bacteria starter fermented in Example 2, put 2ml sterile seawater and 30 Artemia in each hole, and the experimental group is only added Vibrio alginolyticus (10 8 cfu / ml), only add Vibrio alginolyticus antagonistic bacteria starter (10 8 cfu / ml), and Vibrio alginolyticus (10 8 cfu / ml) and Vibrio alginolyticus antagonistic bacteria starter (10 8 cfu / ml), the control group did not add any bacteria, only Artemia. Three parallels were set up in each group, and the conditions were not changed during the experiment. After 4 days of observation, the mortality rate of each group was counted. The specific results are shown in Table 2 and figure 2 show...
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