doi: 10.15389/agrobiology.2023.4.669eng

UDC: 636.5.033:591.1:571.27

Supported financially from the Russian Science Foundation (project № 22-16-00024)


N.V. Bogolyubova ✉, R.V. Nekrasov, D.A. Nikanova,
A.A. Zelenchenkova, N.S. Kolesnik, R.A. Rykov, N.A. Volkova,
A.N. Vetokh, L.A. Ilina

Ernst Federal Research Center for Animal Husbandry,60, pos. Dubrovitsy, Podolsk District, Moscow Province, 142132 Russia, e-mail (✉ corresponding author),,,,,,,,

Bogolyubova N.V.
Rykov R.A.
Nekrasov R.V.
Volkova N.A.
Nikanova D.A.
Vetokh A.N.
Zelenchenkova A.A.
Ilina L.A.
Kolesnik N.S.

Final revision received June 26, 2023
Accepted July 6, 2023

A comparative study of the relationship between the antioxidant protection (AOP) and immunity in poultry of various genotypes is relevant for clinical and physiological assessment of health status and the search for combinations of genotypes to obtain new crosses. In this work, for the first time, differences in biochemical and molecular genetic indicators of antioxidant protection and immunity were established for the Russian White, Ross 308 cross, and Russian White × Cornish cockerels. Correlations were revealed between the expression of some genes for AOP and immunity enzymes in caecum and liver tissues, and the average daily weight gain. The aim of the work was to assess the factors of immunity and antioxidant status, nonspecific immunity indicators, and gene expression levels for enzymes involved in antioxidant protection and immune response in male chickens (Gallus gallus domesticus) of different genotypes. The studies were carried out in 2022 at physiological yard of the Ernst Federal Research Center for Animal Husbandry. Blood samples were taken from Russian White cockerels (RW, n = 28), Ross 308 cross broilers (n = 9) and Russian White × Cornish cockerels (CORN × RW, n = 128) at slaughter at the age of 9 weeks. The TBA test with thiobarbituric acid to measure the TBA-active products (TBA-AP) was performed with Agat-Med kits (Russia). The activity of ceruloplasmin (CP) was determined by the Revin method, the amount of total water-soluble antioxidants (TAWSA) amperometrially (a TsvetYauza-01-AA with an amperometric detector, Khimavtomatika, Russia), the ratio of TBA-AP to the CP was calculated. TAWSA was evaluated as equivalents to gallic acid using calibration solutions with a mass concentration of 0.2, 0.5, 1.0 and 4.0 mg/dm3 prepared from 100 mg/dm3 gallic acid. A solution of orthophosphoric acid (0.0022 mol/dm3)was used as an eluent. Other indicators of antioxidant status were determined with commercial kits (Elabscience Elabscience Biotechnology, Inc., China). Reduced glutathione (E-BC-K096-M), superoxide dismutase (SOD) (E-BC-K020-M), catalase (E-BC-K031-M) and total antioxidant status (TAS) (E-BC-K219-M) were measured by ELISA test (an Immunochem-2100 microplate photometer, High Technology Inc., USA). Nonspecific immunity (i.e., bactericidal activity BA and lysozyme activity LA) of RW (n = 12), CORN × RW (n = 68) and Ross 308 (n = 9) male chicks were determined (a microbiological analyzer Multiskan FC, ThermoFisher Scientific Inc., Finland). Analysis of relative gene expression was performed using real-time PCR. Tissue samples of the caecum and liver were taken from RW (n = 10), Ross 308 (n = 9), and CORN × RW (n = 11) cockerels, 30 samples of each tissue. The relative expression of the genes responsible for antioxidant protection (catalase CAT, glutathione peroxidase GSH-Gpx, heme oxygenase 1 HO-1, superoxide dismutase SOD, related transcription factor 2, NF-E2 Nrf2) and involved in the immune response (avian beta defensin 9 AvBD9, interleukin 6 IL6, interleukin 8 IL8) was assessed. Total antioxidant status (TAS) of broilers was lower than that of analogues, which was confirmed by the maximum content of TBA-AP, 4.27 vs. 3.04 µmol/l for RW (p < 0.05) and 2.79 µmol/l (p < 0.01) for CORN × RW, with a minimum content of ceruloplasmin (37.78 mg/l), and, accordingly, a higher TBA-AP/CP ratio. In the blood of Ross 308 cross males, the maximum TAWSA was detected (49.78 mg/l at p < 0.001 compared to RW), which was due to the maximum amount of reduced glutathione among analogues (38.26 μmol/l at p < 0.001 compared to RW and p < 0.001 compared to CORN × RW). The blood activity of catalase in broilers was also high (100.50 U/l at p < 0.05 compared to RW and p < 0.01 compared to CORN × RW). However, their antioxidant system must work at the maximum to neutralization of reactive oxygen species (ROS). Our data on the expression of AOP and immunity genes confirmed these conclusions. In the caeca of broilers, the genes CAT and GSH-Gpx expression was 5 times higher compared to egg breed cockerels (p = 0.0007 and p = 0.0008, respectively), HO-1 2 times higher ( p = 0.01), SOD higher by 40 %. In the liver of broilers, there was a decrease in the genes SOD and GSH-Gpx expression by 5-6 times compared to RW (p = 0.005 for both genes), CAT expression increased by 27 %, and HO-1 by 42 times (p = 0.001). In broilers, the blood lysozyme concertation and activity were the highest (0.47 μg/ml and 3.14 AU/TP, p < 0.001) with a decrease in the percentage of lysis (36.1 vs. 45.6-48.7 % in other cockerels, p < 0.05) with the minimum BA among analogues. This is confirmed by the fact that the expression of pro-inflammatory cytokines (primarily IL-8) which inhibit humoral immunity was generally lower in the studied broiler tissues while it increased in males of other genotypes. This could lead to a decrease in the humoral response. The average daily weigh gain of poultry highly correlated with the CAT (r = 0.998 at p = 0.03) and AvBD-9 (r = 0.999 at p = 0.016) expression in the caecum. In the caecum, high correlations were found between the expression of CAT and AvBD-9 (r = 0.999 at p = 0.014), IL6 and HO-1 (r = 0.999 at p = 0.1), which confirms the relationship between AOP and bird health. Ross 308 cross broilers showed a higher accumulation of lipid peroxidation products. This highlights the feasibility of using nutritional factors to reduce oxidative stress and increase the antioxidant potential of the body to improve the quality of poultry products.

Keywords: antioxidant status, immunity, chickens, broilers, genotypes, gene expression, CAT, GSH-Gpx, HO-1, SOD, Nrf2, AvBD9, IL6, IL8.



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