Bulletin "Veterinary biotechnology"

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Veterynarna biotekhnolohiia– Veterinary biotechnology, 2023, 43, 62-77 [in Ukrainian]. https://doi.org/10.31073/vet_biotech43-07

KOLESNIKOV A., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it., STEGNII B., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

National Scientific Center «Institute of Experimental and Clinical Veterinary Medicine»

 

DEVELOPMENT OF THE TECHNOLOGICAL REGULATIONS TO PRODUCE AN INACTIVATED VACCINE AGAINST CHICKEN HEMOPHILIA USING UKRAINIAN EPIZOOTIC-RELEVANT STRAINS

Introduction. Both viruses and bacteria, including Avibacterium paragallinarum, formerly known as Haemophilus paragallinarum, can act as etiological agents of respiratory infections in birds. On the territory of Ukraine, nine foreign-made vaccines against hemophiliasis of chickens are registered, which use strains of the pathogen that are relevant for those countries. Therefore, the development of technological regulations to produce an inactivated vaccine using epizootically relevant strains isolated on the territory of Ukraine is relevant.

The goal of the work. To develop a technology for the production of an inactivated emulsified vaccine against hemophilosis of chickens.

Materials and methods. The production of bacterial mass was carried out using epizootically relevant isolates of Av. paragallinarum (SS 6/20, A; SS 7/20, B; SS 8/20, C) in “Brain Heart Infusion” with the addition of V-growth factor (nicotinamide adenine dinucleotide – NAD) and blood serum. Inactivation was carried out using formalin. The following adjuvants were used: “Montanide ISA 70”; aluminum hydroxide (GOA); GOA + saponin. The type of emulsion was determined by the “water drop test” method, stability – by centrifugation (3000 rpm 20 min), and kinematic viscosity using a capillary glass viscometer. The vaccines safety was tested on white mice, 8 heads per group (three experimental and control).

Results of research and discussion. Nutrient medium with the addition of NAD (20 µg/m3) and blood serum (7%) for 18 hours at 37°C under normal atmospheric conditions provided the accumulation of bacterial mass with a concentration of at least 1.0×109 CFU cm3. The optimal inactivant was formaldehyde at a concentration of 0.5% (t 37°C, 48 hours). Three experimental vaccines were obtained using a mixture of antigens (AG) (1:1:1) and adjuvants: sample №1 – AG + “GOA”; №2 – AG + “GOA + saponin”; №3 – AG + “Montanide ISA 70”. The vaccines were stable – from 4% to 8% (requirement: no more than 10%). Viscosity was from 70 mm2/s to 130 mm2/s (requirement: no more than 200 mm2/s). When inoculating the biological material from experimental mice on a nutrient medium, the growth of cultures of Av. paragallinarum was not detected.

 Conclusions and prospects for further research. The development of technological regulations for the production of inactivated emulsified vaccine against hemophilia in chickens is under development. As a result, experimental series of inactivated vaccine against chicken hemophilia using epizootic-relevant Av. paragallinarum strains (SS 6/20, serotype A; SS 7/20, serotype B; SS 8/20, serotype C) were obtained, isolated in Ukraine. The last one complies with the regulatory documents proposed for inactivated vaccines (sterility, stability, viscosity and safety to mice). It is planned to conduct studies on the determination of antigenic and immunogenic activity in poultry. In order to complete the development of technological regulations for vaccine production, it is planned to conduct a series of experiments on the determination of antigenic and immunogenic activity on poultry.

Keywords:pathegen, vaccine, technology, nutrient environment, safety.

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