Use of ethyl lauroyl arginate as nutrition energy substances of feed
A kind of application and feed technology, which is applied in the application field of lauroyl arginine ethyl ester as a feed nutritional energy substance, can solve the problems of untaught and unstudied LAE's single antibacterial effect, reduce risk and improve the survival rate of ducklings , no effect on survival
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Embodiment 1
[0059] Embodiment one: micro-broth dilution method measures LAE to bacterium minimum inhibitory concentration (MIC)
[0060] Principle and purpose: According to the micro broth dilution method stipulated by CLSI, after co-incubating the drug and bacteria in a 96-well plate for 24 hours, the minimum drug concentration at which bacterial growth is inhibited is the minimum inhibitory concentration of the drug.
[0061] Method: LAE and bacteria (the clinical isolate of Riemerella anatipestifer RA-11, Staphylococcus aureus ATCC29213, and Escherichia coli ATCC25922) were diluted with tryptone soybean broth (TSB) and added to In the 96-well plate, a blank control medium CK1 without bacteria, a medium CK2 supplemented with LAE (1000 μg / ml) and a control medium CK3 containing no drug LAE but normal growth of bacteria were set up. The 96-well plate was placed in a 37° C. incubator and incubated for 24 h, and then the absorbance value at 625 nm of each well was measured. OD of blank con...
Embodiment 2
[0066] Embodiment two: LAE is to the mensuration of effective bactericidal time of bacteria
[0067] Principle and purpose: Drugs and bacteria are co-incubated, and some samples are taken out at regular intervals for bacterial plate counting, so as to obtain the growth curve of bacteria under the action of the drug, which is the time-killing curve of the drug.
[0068] Method: Mix the bacteria in the logarithmic phase with different concentrations of LAE solutions, take samples at 0, 0.5, 1, 1.5, and 2 hours after adding the drug, and place 10-fold serial dilutions on the TSA plate, culture at 37°C for 18- 24h. Calculate the bacterial concentration of each dosing group at each time point by the number of colonies, and draw as follows figure 1 Bacterial survival curves are shown.
[0069] Result analysis: at a concentration of 16 μg / ml, LAE can completely kill Riemerella anatipestifer and Escherichia coli within 30 minutes, and reduce the number of Staphylococcus aureus by 6...
Embodiment 3
[0070] Example 3: LAE-induced bacterial drug resistance experiment
[0071] Principle and purpose: Bacteria can be induced to produce drug-resistant bacteria at sub-inhibitory concentrations, and the drug-induced bacteria can be detected by long-term transfer of bacteria cultured with sub-inhibitory concentrations of drugs to fresh drug-containing media Ability to generate resistance mutations.
[0072] Method: The bacteria in the logarithmic phase were mixed with the LAE solution in a glass test tube, sealed with a cotton plug, and placed in a constant temperature shaker at 37°C and 220 rpm for 24 hours. Afterwards, check the MIC of the drug every 24 hours, and add the bacterial solution with the highest drug concentration (that is, the highest drug concentration lower than the MIC) that can cultivate a turbid bacterial solution to fresh cultures containing different drug concentrations at a ratio of 1:100. In Kiri, culture at 37°C, 220rpm for 24h and last for 30 days. Chlo...
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