A tetracycline-resistant Escherichia coli and its application
A technology of Escherichia coli and tetracycline, applied in the biological field, can solve the problems of easy contamination of strains, long time-consuming, cumbersome operation, etc., and achieve good pathogenicity, short time-consuming, and clear-cut effects
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Embodiment 1
[0033] The isolation and identification of embodiment 1 bacteria
[0034] 1. Isolation of bacteria
[0035] (1) Isolation, culture and morphological observation of bacteria
[0036] Take the dead duck liver tissue and aseptically inoculate it on the MacConkey agar plate medium, culture it at a constant temperature of 37°C for 18-24 hours to observe the growth of the bacteria; pick a single colony that turns pink on the medium for Gram staining microscopy, Then pick the same single colony and inoculate it on the MacConkey agar plate medium for further separation and purification, and re-inoculate the isolated and purified bacteria on the agar slant, coded as A, and save it for future use.
[0037] (2), strain A tetracycline susceptibility test
[0038] Strain A was evenly spread and inoculated on the nutrient agar medium, and the tetracycline susceptibility test was carried out according to the K-B disk diffusion method (Shasha, Song Zhenhui. Animal Microbiology Experiment Tu...
Embodiment 2B3
[0060] Example 2B 3 Strain animal regression test
[0061] Will B 3 The strain was inoculated in the nutrient broth medium, shaken and cultured at 37°C for 12 hours, and the bacterial liquid was subjected to McFarland turbidity to make the bacterial liquid content 1×10 8 .cfu / mL (0.5 McFarland concentration). 10 healthy ducklings in the test were divided into 2 groups, respectively, the test group and the control group, 5 in each group; the test group ducklings were intramuscularly injected with 1mL / of the inoculated bacteria solution, and the control group of ducklings were intramuscularly injected with normal saline, 1mL / , and observed 1 week, the incidence and mortality of ducklings were recorded.
[0062] Sensitive bacteria group A and resistant bacteria B 3 The test ducks in the group all showed depression, loss of appetite, unsteady standing, and thin stool after 24 hours of inoculation, died after 36 hours, and all died after 96 hours. Necropsy of dead ducklings s...
Embodiment 3
[0066] In vitro inhibitory effect of embodiment 3 Chinese medicine on tetracycline-resistant B3 bacteria
[0067] (1) Preparation of the tested Chinese medicine solution: take 4 g of five Chinese medicine extracts (each g extract is equivalent to 10 g of the original medicinal material) of Scutellaria baicalensis, rhubarb, Folium Folium, Radix Isatidis and Forsythia, and put them into different beakers respectively. , add 20ml of distilled water, stir to dissolve, filter, and sterilize the filtrate in a vial by steam under reduced pressure (115°C; 20min) for later use. At this time, the concentration of the medicinal solution was 2 g / ml.
[0068] (2) Bacterial solution preparation: inoculate the bacterial strain B3 preserved in glycerol at -20°C on ordinary agar medium, incubate at 37°C for 24 hours, pick typical colonies and inoculate them in ordinary broth medium, and incubate at 37°C for 18-24 hours ,spare.
[0069] (3) Antibacterial test of traditional Chinese medicine b...
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