Bulletin "Veterinary biotechnology"

Veterynarna biotehnologija – Veterinary biotechnology, 2018, 32(1), 169-174 [in Russian]. https://doi.org/10.31073/vet_biotech32(1)-22

NOVIKOVA O.N., LAMAKAY U.V., BELIANKA D.L.e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

S.N. Vyshelesky Institute of Experimental Veterinary Medicine, Minsk, Republic of Belarus


Mannheimia haemolyticа is one of the main agents responsible for fibrinouse pleuropneumonia in young calves and dairy cattle. Mannheimia haemolyticа leukotoxin (Lkt) is the main virulence factor which to play an important role in the protective immunity. Creation of the new vaccines with Lkt in it is the perspective direction to evaluate the efficacy of their using. In order to obtain data concerning the influence the Lkt immunization on the level of antibodies production in g. pigs we used an indirect enzyme-linked immunosorbent assay (ELISA). The procedure of subcutaneous immunization was conducted in guinea pigs with using of three samples: sample №1 concluded only Lkt (positive control –K+), sample № 2 concluded Lkt and two other bacterial antigens–M. haemolytica and P. multocida (experimental vaccine sample +), sample №3 was as a negative control (50% emulsion of Мontanide ISA 206 VG). Paired serum samples were collected from g. pigs on 14 and 21 p.i. day. The antigen for ELISA was a purified in-house preparation of Lkt. For ELISA wells of plates were coated with 0,9 µg Lkt per well, serum samples were diluted 1:100. The data obtained in ELISA were then сalculation. The validity criterias were ∆ОD (К+/К--) ≥ 2,0 and ∆ОD (Sample+/К-) ≥ 2,0. The data ∆ОD (К+/К-) for paired serum sample (№2) were 4,9 and 5,8; ∆ОD (Sample+/К-) – 3,6 and 4,2. The main criteria for level of postvaccinal antibodies in g. pig,s blood was S/P. The results were positive if S/P≥50%. The results S/P for paired serum sample of g. pigs, immunization with sample of the experimental vaccine, were 63,0% and 67,9%. Quantity determination in g. pigs the level of postvaccinal antibodies to M.haemolyticа Lkt is able to conduct the laboratory control of Lkt immunogenic activity as well as provides particularly standartisation of biotechnology process leukotoxin production for vaccine manufacturing.

Keywords: Leukotoxin M. haemolytica, postvaccinal antibodies, guinea pigs, ELISA


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