In Vitro Preparation And Testing Of Anti-Salmonella Vaccine Against Abortion In Sheep

Flore CHIRILA, George Cosmin NADAS, Sorin RAPUNTEAN, Cosmina Maria BOUARI, Septimiu TOADER, Petru BERCE, Andreea SZEKELY, Nicodim Iosif FIT


Abstract: In March 2016, the microbiology laboratory of the Faculty of Veterinary Medicine in Cluj-Napoca, 3 type of antisalmonella vaccine for sheep were prepared. For type 1, 24 hours salmonella on brain heart infusion (BHI) broth culture, heat inactivated for 1 hour at 60°C, then with formaldehyde in a concentration of 3 ‰. Variant 2 - the culture supernatant obtained on solid BHI medium, washed with PBS, frozen-thawed 6 times, centrifuged at 4000 rpm for 15 minutes, filtered through 0.2 μm Millipore filter Orange Scientific. Variant 3 - suspension of cell walls remaining after centrifugation suspended in PBS and inactivated with formaldehyde.

After the bacterial and fungal sterility control, the three types of vaccine were administered in rabbits by the subcutaneous route at a dose of 1 ml/individual along with 1 ml of plant extract adjuvant (decoction of Ewernia prunastri) sterilized by filtration.

There have been two booster inoculations of the initial administration, 7 and 14 days after. Before each dose of vaccine, blood was sampled from the marginal auricular vein in order to control the immunogenicity by anti-somatic ˮOˮ serum antibody (Ab) titration using slow microplate agglutination test (Widal reaction). After three inoculations with the vaccine variant 1, Ab serum titer reached 1/128, and in types 2 and 3, 1/512 after 2 inoculations, decreasing to 1/256 after the second booster administered with no immunomodulator.


Key words: anti-Salmonella vaccine against abortion in sheep, Ewernia prunastri, adjuvant

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