Preparation method for novel adjuvant piglet Escherichia coli inactivated vaccine

[0028] Example 1

[0029] Escherichia coli strains: isolated, identified, screened and conserved by the research team of the Highland Animal Disease Detection Center, College of Animal Science, College of Agriculture and Animal Husbandry, Tibet University.

[0030] Mannitol 2013103101, Chengdu Kelong Chemical; LB medium: Tryptone10g, YeastExtract5g, NaCl10g, with ddH 2 Dilute the volume to 1000ml with O, adjust the pH to 7.0, sterilize by autoclaving, and store at 4°C.

[0031] LB medium plate: Tryptone 10g, YeastExtract 5g, NaCl 10g, agar 20g, fixed to ddH 2 O1000ml, adjust pH to 7.0, autoclave for 20min, pour the dissolved LB medium into a sterilized plate under an alcohol lamp, cool and solidify and store at 4°C for later use.

[0032] Kanamycin (stock solution): Kanamycin 0.3g, dilute to ddH 2 O10ml, filter and sterilize with 0.22um filter, and store at -20℃ for later use.

[0033] Chloramphenicol (storage solution): 0.34g of chloramphenicol, dilute to 10ml of absolute et

[0074] Example 2

[0075] Immunization method:

[0076] The piglets were orally immunized with Escherichia coli inactivated adjuvant vaccine according to the immunization schedule of 0, 14, 28, and 42 days for a total of 4 times. Before each immunization, the vaccine was reconstituted with distilled water and injected intramuscularly: 4ul-40ul per gram of animal body weight The amount of bacteria is 10 6 -10 10 CFU / ml vaccine; oral 1ml-10ml bacteria per kg animal body weight is 10 6 -10 10 CFU / ml vaccine, stirred until the temperature is lower than room temperature and fed to the animals to be immunized.

[0077] Example 3

[0078] (1) Detection of immune effect

[0079] Immunization: Mice were immunized with adjuvanted and unadjuvanted vaccines respectively, and the immunization schedule was performed on 0, 14, 28, and 42 days. Intramuscular injection or oral immunization was used for a total of 4 times. The vaccine was reconstituted with distilled water before each immunization. Immunization by intramuscular injection: 4ul-40ul per gram of animal body weight, and the bacterial volume was 10 6 -10 10 CFU / ml vaccine; Oral immunization: 1-10ml bacterial volume is 10 6 -10 10 The vaccine at CFU / ml was stirred until the temperature was lower than room temperature and fed to the animals to be immunized.