Bulletin "Veterinary biotechnology"

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Veterynarna biotehnologija – Veterinary biotechnology, 2016, 28, 284-294 [in Ukrainian].

TARASOV O., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it., SAPEIKO V., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it., BABKINA M., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

Institute of Veterinary Medicine of NAAS

PINCHUK N., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

State Scientific-Control Institute of Biotechnology and Strains of Microorganisms

INVESTIGATION OF SWINE ERYSIPELAS CAUSATIVE AGENT ANTIGENIC PECULIARITIES

Introduction. Swine erysipelas causes significant losses in Ukraine, the EU and other countries. The study of the antigenic characteristics of the causative agent of erysipelas, the formation of specific immunity and its intensity today does not lose relevance because of antigenic variability that has a direct impact on the effectiveness of preventive vaccination.

Goal of study. To investigate the activity of sera obtained using vaccine strains and the vaccine against swine erysipelas to the antigens of 300, 250, 65–67, 40–45, 25 and 12 kDa. To investigate the protein antigenic specificity of whole cell antigen of erysipelas causative agent using serological tests as well.

Materials and methods of research. Surface antigens were obtained by centrifugation of culture at 4,000 x g for 5 minutes to precipitate the cells. The supernatant was decanted, filtered aseptically through a filter with 0.22 microns diameter of pore. The cells were washed twice with sterile buffer (pH 7.6) containing 0.5% detergent and incubated with shaking for 60 minutes at 37.2±0.3°C. Then the samples were centrifuged at 5000 x g for 5 minutes. The supernatant containing the surface antigens was used in further studies.

The protein composition of antigens were determined by polyacrylamide gel electrophoresis using proposed by U.K. Laemmli (1970), specificity of antigen was confirmed by immunoblot with positive and negative sera by standard protocol.

The evaluation of protein quantities was performed using Bradford standard method.

Sera were obtained by standard protocols.

Results of research and discussion. The antigen specific serum with molecular weight of 300, 250, 65–67,40–45, 25 and 12 kDa, as well as whole cell antigen of vaccine strains and field isolates of the E.rhusiopathiae was obtained. The highest activity of sera obtained using alkaline extract of antigens especially to the 65–67 kDa and 40–45  kDa antigens. In serological tests it was confirmed the antigenic specificity of whole cell antigen.

Established that the maximum activity of mice serum obtained to protein with molecular weight 250 kDa (8.7±0.4 1og2), 65 kDa (8.7±0.3 1og2), 300 kDa (8.3±0.3 1og2). The lowest activity was detected using the protein with molecular weight 12 kDa (titer 6.7±0.2 1og2) and 25 kDa (titer 5.0±0.2 1og2).

For immunization of mice it was used the whole cell antigen of swine erysipelas causative agent prepared in the form of ultrasonic homogenate and alkaline hydrolysate. The proteins of 300, 250, 65–67, 25 and 12 kDa were the most active antigens in specific mice serum that actively respond to ELISA with whole cell antigen. All these proteins induce agglutinating antibodies production in mice.

The results of the study could be used for improving vaccines for swine erysipelas control.

Conclusion.  It was investigate the activity of serum obtained with vaccine strains and the commercially available vaccines against swine erysipelas to the antigens of 300, 250, 65–67, 40-45, 25 and 12 kDa. The maximum activity of mice serum obtained to protein with molecular weight 250 kDa (8.7±0.4 1og2), 65 kDa (8.7±0.3 1og2), 300 kDa (8.3±0.3 1og2). The lowest activity was detected using the protein with molecular weight 12 kDa (titer 6.7±0.2 1og2) and 25 kDa (titer 5.0±0.2 1og2).

Keywords: Erysipelothrix rhusiopathiae, protein, ELISA, antigen

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