Cite: Zhovnir O.M., Tarasov O.A., Kravtsova O.L., Shaposhnik V.M. Antyhenna kompozytsiia ta laboratornyi kontrol eksperymentalnoho zrazka vaktsyny proty bakterioziv ovets [Antigenic composition and laboratory control of experimental sample of vaccine against sheep bacteriosis]. Veterynarna biotekhnolohiia – Veterinary biotechnology, 46, 60-71. https://doi.org/10.31073/vet_biotech46-04 [in Ukrainian].
Zhovnir O.M. 1, e-mail This email address is being protected from spambots. You need JavaScript enabled to view it.
Tarasov O.A. 1, e-mail This email address is being protected from spambots. You need JavaScript enabled to view it.
Kravtsova O.L. 2, e-mail This email address is being protected from spambots. You need JavaScript enabled to view it.
Shaposhnik V.M. 3, e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.
1 Institute of Veterinary Medicine, NAAS of Ukraine, Kyiv, Ukraine
2 State Research Institute for Laboratory Diagnostics and Veterinary and Sanitary Expertise, Kyiv, Ukraine
3 Expert center for diagnostics and laboratory support Biolights LLC
ANTIGENIC COMPOSITION AND LABORATORY CONTROL OF EXPERIMENTAL SAMPLE OF VACCINE AGAINST SHEEP BACTERIOSIS
The presented materials highlight the detailed process of selecting pathogen strains, selecting and establishing their optimal concentrations and determining the ratio between aerobic and anaerobic antigens when creating an experimental sample of an inactivated associated concentrated vaccine. It is known from published data that the main factor in the development of infectious pathology is the pathogenic microbiome of different composition and genesis.
Based on the study of the etiologic structure of infectious disease pathogens, vaccines have been developed in many countries of the world and are used to combat associated infections in many countries, including Ukraine.
The aim of the study was to select a composition of antigens for the design of a prototype of the associated vaccine and to conduct its laboratory control for harmlessness and to determine antigenic activity and immunogenic efficacy.
Materials and methods. It wasused 8 strains of sheep bacteriosis pathogens that were stored in the museum of microorganism strains of the laboratory, namely: C. perfringens type A, C. perfringens type B, C. perfringens type C, C. perfringens type D, C. novyi, C. septicum, P. multocida, Mannheimia haemolytica. All used cultures of microorganisms were previously checked for compliance with morphological, cultural, biochemical and biological properties according to generally accepted methods. After cultivation for 48 hours, the monocultures of pathogens were inactivated with formalin solution and checked for complete inactivation. 2-3 hours before the end of the planned clostridial cultivation period, the cultures were checked for the following indicators: hydrogen ion concentration (pH), growth purity, and the presence of spores. Before the end of the inactivation period, samples were taken from the anacultures and tested for sterility and non-toxicity to white mice.
Laboratory animals were used in the experiment, were treated in accordance with current guidelines and international bioethical standards to minimize animal discomfort and suffering. All procedures met the requirements for the protection of animals used for scientific purposes.
Results of research and discussion. Results of the study and discussion. 8 strains of pathogenic pathogens were selected, consisting of six representatives of anaerobic and 2 aerobic microorganisms. Laboratory studies confirmed: complete inactivation of monocultures of C. perfringens types A, B, C, D, P. multocida, M. haemolytica, the absence of their toxicity, and proven harmlessness for white mice. The concentration of monocultures of pathogens was determined, and the total concentration of microbial cells in the vaccine was established, which was 3.0 × 109 m. cells / cm3. After 10 days of observation, the results of studies of antigenic activity and immunogenic efficacy showed that when the pathogen Cl. perfringens type B was administered to laboratory animals, the survival rate among vaccinated mice was 80.0%, while the death rate among unvaccinated animals was 100.0% of white mice; Cl. perfringens type C – 80.0% survival with a mortality of 100.0%; unvaccinated animals; C. novyi – 100.0% survival with a mortality of 90.0%;
Conclusions and prospects for further research. The completeness of inactivation of monocultures included in the experimental sample of the preparation was confirmed, and their absence of toxicity and harmlessness for white mice and guinea pigs was established. The quantitative content of bacterial cells in 1.0 cm3 of the experimental sample of the preparation was: C. perfringens type A was at the level of 0.54 × 109 m. c.; C. perfringens type B was at the level of 0.44 × 109 m. c.; C. perfringens type C – 0.44 × 109 m. c.; C. perfringens type D was at the level of 0.46 × 109 m. c.; C. novyi – 0.3 × 109 m. c.; C. septicum – 0.37 × 109 m. c.; P. multocida – 0.23 × 109 m. c.; M. haemolytica – 0.22 × 109 m. c. The immunogenic efficacy of the preparation was also proven, since after infection of vaccinated white mice with daily cultures of pathogens C. perfringens type B, C. perfringens type C, C. novyi strain "C - 96", Clostridium septicum strain "Ch - 97", and Pasteurella multocida strain "Polonsky", their survival was 88.0%, while 94.0% of unvaccinated laboratory animals died. The prospect of further research is to verify the efficacy of the experimental vaccine sample on target animals.
Keywords: sheep bacteriosis, vaccine, antigen, laboratory control.
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