doi: 10.15389/agrobiology.2021.2.315eng

UDC: 636.52.58:619:579.62:615.371



M.S. Firsova, А.V. Potekhin, Val.А. Evgrafova, О.V. Pruntova ,
V.S. Rusaleyev, R.V. Yashin

Federal Center for Animal Health Control, FGBU VNIIZZh, mkr. Yurievets, Vladimir, 600901, e-mail,, (✉ corresponding author),,,

Firsova M.S.
Pruntova О.V.
Potekhin А.V.
Rusaleyev V.S.
Evgrafova Val.А.
Yashin R.V.Х

Received September 10, 2020

Avian infectious coryza is caused by the bacteria Avibacterium paragallinarum and occurs throughout the world in countries with a well-developed poultry industry, causing significant economic losses to the poultry industry. The specific prevention is the main link in combating infectious rhinitis of chicken. Vaccination of birds provides expressed immunity due to generation of anti-hemagglutinating antibodies. The presented research study is the first to report on immunobiological properties of two formulations of a developed experimental vaccine against avian infectious coryza which contains the formaldehyde-inactivated antigen of a new A. paragallinarum strain No. 5111 (serogroup B). The aim of the work was to evaluate the safety, antigenic and protective properties of the absorbed and emulsion-based formulations of a vaccine against chicken infectious rhinitis based on the A. paragallinarum strain No. 5111. A whole-cell antigen of the A. paragallinarum strain No. 5111 (serotype B-1) inactivated with formaldehyde was used to produce experimental samples of the vaccine for trials. A dose for immunization (0,5 cm3) contained 109 inactivated microbial cells and 3.75 mg of aluminum hydroxide for the absorbed vaccine formulation or oil adjuvant Montanide ISA 70 VG («SEPPIC», France, 70 % wgt) for the emulsion-based formulation. The immunobiological properties of the vaccine were tested on 125 Haysex brown chickens (Gallus gallus L.) of 1.5-2.0 months of age which were seronegative to A. paragallinarum. The safety of the vaccine samples was tested by injecting chickens in a 2-fold dose (1.0 cm3). Each sample was injected subcutaneously in the middle third of the neck and intramuscularly in the chest using 5 chickens per each formulation. The clinical status of the birds was observed daily for 42 days. At the end of the experiment, the chickens were slaughtered and the incision of the injection site was visually examined. Three groups of chickens (25 birds each, 75 chickens in total) were assigned to determine protective properties of the vaccine. The birds of group I were immunized with the absorbed formulation of the vaccine, the chickens of group II were injected with the emulsion-based formulation. The birds were injected subcutaneously into the middle third of the neck at a dose of 0.5 cm3 twice with a 20-day interval. Unvaccinated chickens of group III were used as a control. In 15 days after revaccination, the chickens of groups I, II, and III were infected with a 1-day broth culture of the A. paragallinarum strain No. 5111 with a concentration of 5 units according to the optical standard of bacterial suspension turbidity. The clinical status of the chickens was observed during 7 days after infection. The post-mortem examination was performed with a bacteriological analysis of the contents of the nasal sinuses during the experiment and at the end of the experiment. The vaccine antigenicity and the duration of immunity were determined on 30 birds (three groups of 10 birds each). The chickens of group I were immunized with the absorbed vaccine sample, group II — with an emulsion-based sample, and unvaccinated birds of group III served as a control. The vaccine antigenicity was assessed based on humoral antibody level using the hemagglutination inhibition test (HI test). The mild to moderate tissue lesions were observed at the injection site without an obvious inflammatory reaction. For the absorbed formulation, slight subcutaneous swellings and hyperemia were observed in some chickens at the injection site. For the emulsion-based formulation, the formation of connective tissue granules with the vaccine residues without necrotic lesions and an obvious inflammatory reaction of the surrounding tissues occurred at the injection site in all birds. No significant differences in the condition of chicks from vaccinated groups were observed (p > 0.05), but there was a significant difference between the birds of the test and control groups (p < 0.05). The level of protection of chickens after double immunization with the adsorbed vaccine and the emulsion-based vaccine was 92 % and 88 %, respectively. Twenty days after the first vaccination with absorbed and emulsion-based formulations, the average antibody titers were below the threshold level (p > 0.05). Increased antibody titers in chicken sera were observed only at day 15 post the second immunization. At day 60 post vaccination, the antibody levels in the chicken sera reached their maximum, i.e., 7.5±0.8 log2 in poultry immunized with the adsorbed vaccine and 8.9±0.7 log2 in birds immunized with the emulsion-based vaccine (p > 0.05). In chickens vaccinated with the vaccine containing aluminum hydroxide gel, a decrease in the antibody titer to 5.5±0.7 log2 was observed at day 240 while in birds immunized with the emulsion-based vaccine the titer remained at the level of 8.7±0.8 log2 (p > 0.05). No specific antibodies to the causative agent of infectious coryza were detected in chickens of the control group during the entire observation period, including the diseased and convalescence period. Thus, our findings show that the adsorbed and emulsion-based experimental formulations of the developed vaccine against avian infectious coryza are safe and demonstrate high antigenicity and immunogenicity after double administration.

Keywords: infectious coryza in chicken, Avibacterium paragallinarum strains, antigen, adjuvant, sample of vaccine.



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