Glycine-glucose-exopolysaccharide compound feed additive
A feed additive and exopolysaccharide technology, which is applied in animal feed, pharmaceutical formula, additional food elements, etc., to achieve the effect of improving sensitivity, improving immunity and body function
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Embodiment 1
[0047]Determination of drug resistance of Edwardsiella tarda EIB202
[0048] Edwardsiella tarda is a Gram-negative short bacillus. It was first reported by Hoshina (1 96 2a), and it is related to reddisease of Japanese eel. From the first report to now, the fungus has caused diseases in more than 20 kinds of fish, such as eel, flounder, tilapia, Chinese soft-shelled turtle, carp, etc., causing huge losses to aquaculture. Edwardsiella tarda is also a common pathogenic bacteria in humans and fish, which directly poses a threat to human health.
[0049] Firstly, the minimum inhibitory concentration of Edwardsiella tarda to various antibiotics was determined. The results showed that the minimum inhibitory concentration of Edwardsiella tarda EIB202 to kanamycin was 12.5 μg / mg, the minimum inhibitory concentration to tetracycline was 125 μg / mg, and the minimum inhibitory concentration to chloramphenicol was 50 μg / mg, indicating that Edwardsiella tarda EIB202 is a multidrug resist...
Embodiment 2
[0051] Glycine increases the susceptibility of Edwardsiella tarda to kanamycin
[0052] 1. Preparation of test samples: Pick a single colony of Edwardsiella tarda EIB202 from the LB plate and inoculate it in 5 mg of LB medium, culture at 200 rpm at 30°C for 24 hours to reach saturation. Collect the bacterial liquid by centrifugation, centrifuge at 8000rpm for 5min, remove the supernatant and wash the bacterial cells with 0.85% normal saline, and finally suspend the bacterial cells with 1×M9 (containing 10mM acetate), adjust the OD value of the bacterial solution to 0.2, and then pack them separately 5mL in a test tube for later use.
[0053] 2. Glycine can increase the sensitivity of Edwardsiella tarda EIB202 to kanamycin
[0054] In order to know whether glycine contributes to the bactericidal effect of bacteria, the experiment of adding glycine under the premise of adding kanamycin was carried out. To the above prepared bacterial samples, first add 40 μg / mL kanamycin, then...
Embodiment 3
[0056] Glucose synergizes with glycine to increase susceptibility of Edwardsiella tarda
[0057] It has been reported in the literature that glucose can increase the sensitivity of aminoglycosides to Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) persistent drug-resistant bacteria (persistence), and it has been proved that this phenomenon is the Unique to glycoside antibiotics. We therefore also investigated the susceptibility of bacteria to kanamycin for glucose, and glycine and glucose.
[0058] 1. Glucose and glycine can synergistically increase the susceptibility of Edwardsiella tarda to kanamycin
[0059] In order to study whether glucose and glucose can cooperate with glycine to increase the sensitivity of bacteria to kanamycin, the experiment was divided into three groups: one group only added 40 μg / mL kanamycin in the prepared samples as a control, and the other two groups On the basis of adding 40 μg / mL kanamycin, 10 mM glucose, 10 mM glu...
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