Rough type Brucella melitensis attenuated strain and vaccine thereof
A kind of technology of Brucella sheep and rough type, applied in the field of rough type Brucella sheep attenuated strain and vaccine field
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Embodiment 1
[0065] Pancreatic Soy Broth (TSB, BD Company, USA) or other suitable culture medium is used as the culture medium for the rough Brucella vaccine strain (RA343). Inject 1% to 2% of the amount into the seed liquid of Brucella melitansis RM57 strain, gradually increase the ventilation at 36-37°C, and ferment and cultivate for 36 hours according to the conventional method. During the cultivation process, you can add For 50% glucose solution, the amount added each time is 1% to 2% of the total amount of the medium.
[0066] After the cultivation is completed, try to use pancreatic soybean agar medium for sampling to conduct a pure test according to the method stipulated in the "Chinese Veterinary Pharmacopoeia", and it is pure. At the same time, take samples and count live bacteria with TSA for reference during concentration.
[0067] Add 0.1% to 0.2% of the total amount of sodium carboxymethyl cellulose to the bacteria liquid that has passed the pure inspection to precipitate the...
Embodiment 2
[0072] Product testing
[0073] The relevant inspection methods involved in the present invention are carried out according to the method of "Chinese Veterinary Pharmacopoeia"
[0074] (1) Physical properties The vaccine should be loose and fast, and can dissolve rapidly within 1 minute after adding the diluent;
[0075] (2) The purely tested vaccines are purely free of other bacteria;
[0076] (3) Safety inspection Dilute the vaccine to contain 1×10 per 1ml 9 CFU live bacteria, subcutaneously inoculate 5 Kunming mice of 18-20 g in the groin, each 0.25 ml / only, observe for 6 days, all of them are healthy and alive (results are shown in Table 1);
[0077] Table 1 Safety inspection results of live brucellosis vaccine (RM57 strain)
[0078]
[0079] -: Healthy survival, no abnormality at the injection site; +: Death or morbidity.
[0080] (4) Viable bacteria count Use TSA medium to count the viable bacteria to count the viable bacteria of the vaccine, and the number of via...
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