PLANT BIOLOGY
ANIMAL BIOLOGY
SUBSCRIPTION
E-SUBSCRIPTION
 
MAP
MAIN PAGE

 

 

 

 

Bogolyubova N.V., Nekrasov R.V., Nikanova D.A., Zelenchenkova A.A., Kolesnik N.S., Rykov R.A., Volkova N.A., Vetokh A.N., Ilina L.A.. Biochemical and molecular genetic indicators of antioxidant protection and immunity in male chicks (Gallus gallus domesticus) of different genotypes. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2023, Vol. 58, № 4, p. 669-684.
(how to cite this article)

doi: 10.15389/agrobiology.2023.4.669eng

UDC: 636.5.033:591.1:571.27

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

BIOCHEMICAL AND MOLECULAR GENETIC INDICATORS OF ANTIOXIDANT PROTECTION AND IMMUNITY IN MALE CHICKS (Gallus gallus domesticus) OF DIFFERENT GENOTYPES

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 652202@mail.ru (✉ corresponding author), nek_roman@mail.ru, dap2189@gmail.com, aly4383@mail.ru, kominisiko@mail.ru, brukw@bk.ru, natavolkova@inbox.ru, anastezuya@mail.ru, ilina@biotrof.ru

ORCID:
Bogolyubova N.V. orcid.org/0000-0002-0520-7022
Rykov R.A. orcid.org/0000-0003-0228-8901
Nekrasov R.V. orcid.org/0000-0003-4242-2239
Volkova N.A. orcid.org/0000-0001-7191-3550
Nikanova D.A. orcid.org/0000-0001-5164-244X
Vetokh A.N. orcid.org/0000-0002-2865-5960
Zelenchenkova A.A. orcid.org/0000-0001-8862-3648
Ilina L.A. orcid.org/0000-0003-2789-4844
Kolesnik N.S. orcid.org/0000-0002-4267-5300

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.

 

REFERENCES

  1. Bogolyubova N.V., Nekrasov R.V., Zelenchenkova A.A. Antioxidant status and quality of poultry and animal meat under stress and its correction with the use of various adaptogens (review). Sel'skokhozyaistvennaya biologiya [Agricultural Biology], 2022, 57(4): 628-663 CrossRef
  2. Xing T., Gao F., Tume R.K., Zhou G., Xu X. Stress effects on meat quality: a mechanistic perspective. Comprehensive Reviews in Food Science and Food Safety, 2019, 18(2): 380-401 CrossRef
  3. Zhang W., Xiao S., Lee E.J., Ahn D.U. Consumption of oxidized oil increases oxidative stress in broilers and affects the quality of breast meat. J. Agric. Food Chem., 2011, 59(3): 969-974 CrossRef
  4. Sihvo H.K., Immonen K., Puolanne E. Myodegeneration with fibrosis and regeneration in the pectoralis major muscle of broilers. Veterinary Pathology, 2014, 51(3): 619-623 CrossRef
  5. Surai P.F., Kochish I.I., Fisinin V.I., Kidd M.T. Antioxidant defence systems and oxidative stress in poultry biology: an update. Antioxidants, 2019, 8(7): 235 CrossRef
  6. Estévez M. Oxidative damage to poultry: from farm to fork. Poultry Science, 2015, 94(6): 1368-1378 CrossRef
  7. Belhadj Slimen I., Najar T., Ghram A., Abdrrabba M.J.O.A.P. Heat stress effects on livestock: molecular, cellular and metabolic aspects, a review. Journal of Animal Physiology and Animal Nutrition, 2016, 100(3): 401-412 CrossRef
  8. Wen C., Chen Y., Leng Z., Ding L., Wang T., Zhou Y. Dietary betaine improves meat quality and oxidative status of broilers under heat stress. Journal of the Science of Food and Agriculture, 2019, 99(2): 620-623 CrossRef
  9. Wein Y., Bar Shira E., Friedman A. Avoiding handling-induced stress in poultry: use of uniform parameters to accurately determine physiological stress. Poultry Science, 2017, 96(1): 65-73 CrossRef
  10. Radi R. Oxygen radicals, nitric oxide, and peroxynitrite: redox pathways in molecular medicine. Proceedings of the National Academy of Sciences, 2018, 115(23): 5839-5848 CrossRef
  11. Ludin A., Gur-Cohen S., Golan K., Kaufmann K.B., Itkin T., Medaglia C., Lu X.J., Ledergor G., Kollet O., Lapidot T. Reactive oxygen species regulate hematopoietic stem cell self-renewal, migration and development, as well as their bone marrow microenvironment. Antioxidants & Redox Signaling, 2014, 21(11): 1605-1619 CrossRef
  12. Mavangira V., Sordillo L.M. Role of lipid mediators in the regulation of oxidative stress and inflammatory responses in dairy cattle. Research in Veterinary Science, 2018, 116: 4-14 CrossRef
  13. Mesa-Garcia M.D., Plaza-Diaz J., Gomez-Llorente C. Molecular basis of oxidative stress and inflammation. In: Obesity. A.M. del Moral, C.M.A. García (eds.). Academic Press, Sandiego, 2018: 41-62.
  14. McCarthy C.G., Saha P., Golonka R.M., Wenceslau C.F., Joe B., Vijay‐Kumar M. Innate immune cells and hypertension: neutrophils and neutrophil extracellular traps (NETs). Comprehensive Physiology, 2021, 11(2): 1575-1589 CrossRef
  15. Ratnam D.V., Ankola D.D., Bhardwaj V., Sahana D.K., Kumar M.N. Role of antioxidants in prophylaxis and therapy: a pharmaceutical perspective. Journal of Controlled Release, 2006, 113(3): 189-207 CrossRef
  16. Kotarev V.I., Alekhin Yu.N., Dolgopolov V.N. Veterinarnyy farmakologicheskiy vestnik, 2017, 1(1): 73-79 (in Russ.).
  17. Mattioli S., Cartoni Mancinelli A., Menchetti L., Dal Bosco A., Madeo L., Guarino Amato M., Moscati L., Cotozzolo E., Ciarelli C., Angelucci E., Castellini C. How the kinetic behavior of organic chickens affects productive performance and blood and meat oxidative status: a study of six poultry genotypes. Poultry Science, 2021, 100(9): 101297 CrossRef
  18. Lengkidworraphiphat P., Wongpoomchai R., Taya S., Jaturasitha S. Effect of genotypes on macronutrients and antioxidant capacity of chicken breast meat. Asian-Australasian Journal of Animal Sciences, 2020, 33(11): 1817-1823 CrossRef
  19. Kondrakhin I.P., Arkhipov A.V., Levchenko V.I., Talanov G.A., Frolova L.A., Novikov V.Е. Metody veterinarnoy klinicheskoy laboratornoy diagnostiki [Methods of veterinary clinical laboratory diagnostics]. Moscow, 2004 (in Russ.).
  20. Voronin E.S., Petrov A.M., Serykh M.M., Devrishov D.A. Immunologiya /Pod redaktsiey E.S. Voronina [Immunology. E.S. Voronin (ed.)]. Moscow, 2002 (in Russ.).
  21. Solovykh G.N., Minakova V.V., Korobov V.P., Lemkina L.M., Karnaukhova I.V., Ryabtseva E.A., Kanunikova E.A. Sposob opredeleniya lizotsimnoy aktivnosti biologicheskikh ob’ektov. Patent RU 2294373C2. 2294373C2 (RF) GOU VO «Orenburgskaya gosudarstvennaya meditsinskaya akademiya» (RF). № 2005103265/13A. Zayavl. 08.02.2005. Opubl. 27.02.2007 [Method for determining lysozyme activity of biological objects. Patent RU 2294373C2. 2294373C2 (RF) State Educational Institution of Higher Education "Orenburg State Medical Academy" (RF) № 2005103265/13A. MPK G01N33/48 A61D99/00. FGBOU VO «SPBGAVM» (RF). № 2010153464/13. Appl. 08.02.2005. Publ. 27.02.2007. Bull. № 1] (in Russ.).
  22. Meza Cerda M.-I., Gray R., Higgins D.P. Cytokine RT-qPCR and ddPCR for immunological investigations of the endangered Australian sea lion (Neophoca cinerea) and other mammals. PeerJ, 2020, 8: e10306 CrossRef
  23. Laptev G.Y., Filippova V.A., Kochish I.I., Yildirim E.A., Ilina L.A., Dubrovin A. V, Brazhnik E.A., Novikova N.I., Novikova O.B., Dmitrieva M.E., Smolensky V.I., Surai P.F., Griffin D.K., Romanov M.N. Examination of the expression of immunity genes and bacterial profiles in the caecum of growing chickens infected with Salmonella enteritidis and fed a phytobiotic. Animals, 2019, 9(9): 615 CrossRef
  24. Livak K.J., Schmittgen T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods, 2001, 25(4): 402-408 CrossRef
  25. Vladimirov Yu.A., Archakov A.I. Perekisnoe okislenie lipidov v biologicheskikh membranakh [Lipid peroxidation in biological membranes]. Moscow, 1972 (in Russ.).
  26. Kapuy O., Papp D., Vellai T., Banhegyi G., Korcsmaros T. Systems-level feedbacks of NRF2 controlling autophagy upon oxidative stress response. Antioxidants, 2018, 7(3): 39 CrossRef
  27. Makhanova R.S. Izvestiya Orenburgskogo gosudarstvennogo agrarnogo universiteta, 2011, 1(29-1): 231-234 (in Russ.).
  28. Całyniuk B., Grochowska-Niedworok E., Walkiewicz K.W., Kawecka S., Popiołek E., Fatyga E. Malondialdehyde (MDA)—product of lipid peroxidation as marker of homeostasis disorders and aging. Annales Academiae Medicae Silesiensis — Śląski Uniwersytet Medyczny w Katowicach, 2016, 70: 224-228 CrossRef
  29. Linder M.C. Ceruloplasmin and other copper binding components of blood plasma and their functions: an update. Metallomics, 2016, 8(9): 887-905 CrossRef
  30. Wu R., Feng J., Yang Y., Dai C., Lu A., Li J., Liao Y., Xiang M., Huang Q., Wang D., Du X. Significance of serum total oxidant/antioxidant status in patients with colorectal cancer. PLoS ONE, 2017, 12(1): e0170003 CrossRef
  31. Gorlov I.F., Tikhonov S.L., Tikhonova N.V. Industriya pitaniya, 2016, 1: 44-53.
  32. Pigeolet E., Corbisier P., Houbion A., Lambert D., Michiels C., Raes M., Zachary M.D., Remacle J. Glutathione peroxidase, superoxide dismutase, and catalase inactivation by peroxides and oxygen derived free radicals. Mechanisms of Ageing and Development, 1990, 51(3): 283-297 CrossRef
  33. Madkour M., Aboelazab O., Abd El-Azeem N., Younis E., Shourrap M. Growth performance and hepatic antioxidants responses to early thermal conditioning in broiler chickens. Journal of Animal Physiology and Animal Nutrition, 2023, 107(1): 182-191 CrossRef
  34. Ostrenko K.S., Galochkina V.P. Veterinariya, 2020, 11: 53-58 (in Russ.).
  35. Stoyanovskyy V.G., Krogh A.O., Kolomiіets I.A. Adaptation of the status of non-specific resistance of the ducks organism in stress conditions inclusion in the ration of probiotical additives. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies, 2018, 20(87): 32-37 CrossRef
  36. Madej J.P., Skonieczna J., Siwek M., Kowalczykc A., Łukaszewicz E., Slawinska A. Genotype-dependent development of cellular and humoral immunity in the spleen and cecal tonsils of chickens stimulated in ovo with bioactive compounds. Poultry Science, 2020, 99(9): 4343-4350 CrossRef
  37. Van Phi L. Transcriptional activation of the chicken lysozyme gene by NF-kappa Bp65 (RelA) and c-Rel, but not by NF-kappa Bp50. Biochem. J., 1996, 313(1): 39-44 CrossRef
  38. Ovsyannikov V.G., Alekseev V.V., Boychenko A.E., Labushkina A.V., Alekseeva N.S., Abramova M.V., Alekseeva N.A. Zhurnal fundamental’noy meditsiny i biologii, 2015, 4: 4-13 (in Russ.).
  39. Gulati K., Guhathakurta S., Joshi J., Rai N., Ray A. Cytokines and their role in health and disease: a brief overview. MOJImmunol., 2016, 4(2): 00121 CrossRef
  40. Ershov F.I. Vestnik RAMN, 2006, 9-10: 45-50 (in Russ.).
  41. Fisinin V.I., Mityushnikov V., Kravchenko N. Ptitsevodstvo, 1977, 7: 28-30 (in Russ.).
  42. Song B., Tang D., Yan S., Fan H., Li G., Shahid M.S., Mahmood T., Guo Y. Effects of age on immune function in broiler chickens. J. Animal Sci. Biotechnol., 2021, 12: 1-12 CrossRef
  43. Surai P.F., Fisinin V.I. Vitagenes in poultry production: Part 1. Technological and environmental stresses. World's Poultry Science Journal, 2016, 72(4): 721-733 CrossRef
  44. Miftakhutdinov A.V. Experimental approaches to stress diagnostics in poultry (review). Sel'skokhozyaistvennaya biologiya [Agricultural Biology], 2014, 2: 20-30 CrossRef
  45. Surai P.F. Antioxidant systems in poultry biology: superoxide dismutase. Journal of Animal Research and Nutrition, 2016, 1(1): 8 CrossRef
  46. Maamoun H., Benameur T., Pintus G., Munusamy S., Agouni A. Crosstalk between oxidative stress and endoplasmic reticulum (ER) stress in endothelial dysfunction and aberrant angiogenesis associated with diabetes: a focus on the protective roles of heme oxygenase (HO)-1. Front. Physiol., 2019, 10: 70 CrossRef
  47. Waza A.A., Hamid Z., Ali S., Bhat S.A., Bhat M.A. A review on heme oxygenase-1 induction: Is it a necessary evil. Inflamm. Res., 2018, 67: 579-588 CrossRef
  48. He F., Ru X., Wen T. NRF2, a transcription factor for stress response and beyond. International Journal of Molecular Sciences, 2020, 21(13): 4777 CrossRef
  49. Sarapul’tsev P.A., Sarapul’tsev A.P. Stress i immunnaya sistema. Tsitokiny i vospalenie, 2014, 3(4): 5-10 (in Russ.).
  50. Li Y., Ma Q.-G., Zhao L.-H., Wei H., Duan G.-X., Zhang J.-Y., Ji C. Effects of lipoic acid on immune function, the antioxidant defense system, and inflammation-related genes expression of broiler chickens fed aflatoxin contaminated diets. InternationalJournalofMolecularSciences, 2014, 15(4): 5649-5662 CrossRef
  51. Narushin V.G., Selina M.V., Romanov M.N. MaterialyMezhdunarodnoynauchno-prakticheskoykonferentsii «Molekulyarno-geneticheskie tekhnologii dlya analiza еkspressii genov produktivnosti i ustoychivosti k zabolevaniyam zhivotnykh» [Proc. Int. Conf. «Molecular genetic analysis of expression of genes for productivity and resistance to animal diseases»]. Moscow, 2019: 67-82 (in Russ.).

 

back