Aug 13, 2020
Salmonella live attenuated vaccine preparation.
- Angel A Justiz-Vaillant1,
- Suzette Curtello2
- 1University of the West Indies St. Augustine;
- 2University of the West Indies. Mona Campus.
- University of the West Indies
- angel.vaillant@sta.uwi.edu
Protocol Citation: Angel A Justiz-Vaillant, Suzette Curtello 2020. Salmonella live attenuated vaccine preparation.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjpdkmi6
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 13, 2020
Last Modified: August 13, 2020
Protocol Integer ID: 40389
A Salmonella cocktail (mixture of five Salmonella strains: S. Montevideo, S. Yeerongpilly, S. Augustenborg, S. Kentucky and S.Typhimurium) is incubated at 37°C in peptone broth with combinations of natural extracts.
Preparation includes clean, sun dried onion and garlic cloves.
After a week of drying, onion and the cloves of garlic is separately grounded using electric blender,
Dilute the plant preparations in 1L distilled water.
Each extract is used individually and in combined concentration.
Five ml of the diluted extract (15 mg/ml) is incorporated in differential and selective Salmonella plating media.
A bacteriogram is carried out to check for bacteria attenuation.
The immunogen contains 5 serovars of Salmonella (10^8 CFU of vaccine strain) per chicken in 1 ml of incomplete Freund’s adjuvant.
It is given intramuscularly at chicken breast
The immunogen (5 serovars) without incomplete Freund’s adjuvant (IFA) is given orally (10^8 CFU per chicken).
All Salmonella isolates were investigated for their antibiotic resistance with the disc diffusion test using the following discs (Difco):
gentamicin (10 μg), kanamycin (30 μg), ampicillin (10 μg), amikacin (30 μg), trimethoprim/sulfamethoxazole (1.25/23.75 μg), chloramphenicol (30 μg), cefazolin (30 μg), cephalothin (30 μg), cefepime (30 μg), cefotaxime (30 μg), streptomycin (10 μg), cefoxitin (30 μg), nalidixic acid (30 μg), ciprofloxacin (5 μg), norfloxacin (10 μg), tetracycline (30 μg ) and imipenem (10 μg).
The five Salmonella serovars were fully attenuated and inoculated in appropriate media and store at -20 degree Celsius.