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Yildirym E.A., Ilyina L.A., Laptev G.Yu., Filippova V.A., Tyurina D.G., Sokolova K.A., Zaikin V.A., Ponomareva E.S., Fisinin V.I., Egorov I.A., Egorova T.A., Manukyan V.A., Lenkova T.N., Degtyareva O.N., Tishenkova M.S., Demidova E.S., Kashporov L.M. , Pashchenko V.E. Changes of gene expression in male and female Smena 9 broiler chickens (Gallus gallus) fed dietary betaine. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2025, Vol. 60, № 6, p. 1097-1112.
(how to cite this article)

doi: 10.15389/agrobiology.2025.6.1097eng

UDC: 619:636.52/.58:636.084:577.2

Acknowledgements:
Supported financially by the Russian Science Foundation, grant No. 22-66-00061, https://rscf.ru/project/22-66-00061/.

 

CHANGES OF GENE EXPRESSION IN MALE AND FEMALE SMENA 9 BROILER CHICKENS (Gallus gallus) FED DIETARY BETAINE

E.A. Yildirym1, 2 ✉, L.A. Ilyina1, 2, G.Yu. Laptev1, V.A. Filippova1, 2,
D.G. Tyurina1, K.A. Sokolova1, 2, V.A. Zaikin1, E.S. Ponomareva1,
V.I. Fisinin3, I.A. Egorov3, T.A. Egorova3, V.A. Manukyan3,
T.N. Lenkova3, O.N. Degtyareva3, M.S. Tishenkova3, E.S. Demidova3,
L.M. Kashporov3, V.E. Pashchenko3

1JSC Biotrof+, 19, korp. 1, Zagrebskii bulv., St. Petersburg, 192284 Russia, e-mail ilina@biotrof.ru, laptev@biotrof.ru,
tiurina@biotrof.ru, dfcx@biotrof.ru, kate@biotrof.ru;
2Saint Petersburg State Agrarian University, 2, Sankt-Peterburgskoe sh., St. Petersburg, 196601 Russia, e-mail deniz@biotrof.ru (✉ corresponding author), ilippova@biotrof.ru, ksenya.k.a@biotrof.ru;
3Federal Scientific Center All-Russian Research and Technological Poultry Institute RAS, 10, ul. Ptitsegradskaya, Sergiev Posad, Moscow Province, 141311 Russia, e-mail olga@vnitip.ru, eta164@yandex.ru, manukyan@vnitip.ru,
dissоvet@vnitip.ru, fncvnitip@mail.ru, tishenkova.m@yandex.ru,
mixalysha@mail.ru, lev-lud2@live.ru, viktoriia_pashchenko@mail.ru

ORCID:
Yildirym E.A. orcid.org/0000-0002-5846-5105
Egorov I.A. orcid.org/0000-0001-9122-9553
Ilyina L.A. orcid.org/0000-0003-2789-4844
Egorova T.A. orcid.org/0000-0002-5102-2248
Laptev G.Yu. orcid.org/0000-0002-8795-6659
Manukyan V.A. orcid.org/0000-0003-4564-4427
Filippova V.A. orcid.org/0000-0001-8789-9837
Lenkova T.N. orcid.org/0000-0001-8391-5000
Tyurina D.G. orcid.org/0000-0001-9001-2432
Degtyareva O.N. orcid.org/0000-0001-7243-7381
Sokolova K.A. orcid.org/0000-0002-9541-6839
Tishenkova M.S. orcid.org/0000-0002-2911-5640
Zaikin V.A. orcid.org/0009-0006-8029-9955
Demidova E.S. orcid.org/0000-0002-0108-2218
Ponomareva E.S. orcid.org/0000-0002-4336-8273
Kashporov L.M. orcid.org/0009-0000-5100-4843
Fisinin V.I. orcid.org/0000-0003-0081-6336
Pashchenko V.E. orcid.org/0000-0001-7484-196X

Final revision received September 26, 2025
Accepted November 03, 2025

Optimizing poultry diets remains a pressing issue in modern poultry farming, given the desire to increase production profitability while minimizing the cost of expensive diet components. This study provides the first results demonstrating the effect of betaine supplementation on the expression of key genes associated with immunity, inflammation, intestinal barrier function, antioxidant activity, and nutrient transport in Smena 9 broiler chickens fed diets with reduced metabolizable energy (by 5%), lysine (by 10 %), and methionine (by 10 %). The work aimed at studying effects of 200-400 g/t betaine on the expression of a number of genes in the caecum tissues in Smena 9 cross broiler chickens/ In addition, we aimed to assess its effect on the meat productivity of poultry fed a diet with a reduced content of metabolizable energy, lysine and methionine. The study was conducted on broiler chickens (Gallus gallus) of the Smena 9 cross (Zagorskoye EPH, the All-Russian Research and Technological Poultry Institute RAS, Sergiev Posad, 2025). Four groups of birds were formed (n = 36 each), equal in males and females. Group I (control) received the basic diet (BD) with a reduced content of metabolizable energy (by 5 %), lysine (by 10 %) and 200 g/t of betaine crystalline powder containing 95 % trimethylglycine (Taian Havay Chemicals Co., Ltd, China). OR in group III was added with 300 g/t of betaine, in group IV with 400 g/t of betaine. Betaine was added by thoroughly mixing it with the compound feed manually. Feeding was ad libitum using crumbled compound feed. The feeding cycle was three-stage, the first stage (up to 14 days of life) with the starter compound feed Start (SGC Zagorskoye EPH, Russia); the second stage (15-21 days) with growth-forming compound feed Rost (SGC Zagorskoe EPH, Russia), and the third stage (22-35 days) with the finishing compound feed Finish (SGC Zagorskoe EPH, Russia). At the end of the experiment, the birds were decapitated and the caeca tissues were collected for gene expression analysis. Gene expression analysis was performed by quantitative real-time polymerase chain reaction (qRT-PCR). Total RNA was isolated from tissue samples using the commercial Aurum™ Total RNA kit (Bio-Rad, USA) according to the manufacturer's protocol. qRT-PCR was performed (a DTlite 4S1 detection amplifier, NPK DNA-technology, Russia) using the SsoAdvanced™ Universal SYBR® Green Supermix kit (Bio-Rad, USA). Primers were used to the genes IL6, IL8L2, PTGS2, encoding proinflammatory cytokines, AvBD1, AvBD2, AvBD9, AvBD10, AvBD11, encoding antimicrobial peptides, Casps6, encoding caspase-6, MUC2, encoding mucin synthesis, SGLT2, encoding sodium-glucose cotransporter 2, SOD1, encoding superoxide dismutase 1. Relative gene expression was calculated by the 2-ΔΔCTmethod. The chickens’ bodyweight, average daily live weight gain, slaughter yield and abdominal fat content in carcasses were assessed individually. The most significant increase in the live weight of cockerels (by 7.2 and 7.3 %, p  ≤ 0.05) was observed at higher doses betaine (300 and 400 g/t), demonstrating the previously known properties of betaine in improving the productivity of Smena 9 broiler chickens. We also revealed a significant decrease in abdominal fat deposition (p ≤ 0.05). Betaine caused a differentiated response characterized by an increase in the expression of the SOD1 gene, a key gene for antioxidant defense. The effect of betaine on the expression of many immune genes depended on the sex of the bird. Thus, in hens, there was a more than 8-fold increase in the expression of the AvBD1 gene for 200 g/t betaine (p ≤ 0.01). At higher doses, the expression of other genes of the AvBD family decreased (p ≤ 0.05). A decrease in the mRNA level of proinflammatory genes IL6, IL8, and PTGS2 (p ≤ 0.05) occurred. The Casp6 mRNA level increased in hens fed 200 g/t betaine by 13.9 times compared to the control (p ≤ 0.01). In cockerels of the group fed 200 g/t betaine, there was a sharp jump in the expression of SGLT2 (sodium-glucose transporter type 2) by 362 times (p ≤ 0.001), with an increase by 39.4 and 13.5 times for 300 and 400 g/t, respectively (p ≤ 0.01). Thus, when metabolizable energy, lysine and methionine in feeds are reduced, the dietary betaine has a positive effect on the production indicators of Smena 9 cross broiler chickens. The results obtained demonstrate complex and sex- and dosage-dependent effects of betaine on the immune system of the birds, as well as its potential role in modulating inflammatory processesand improving glucose metabolism through the regulation of gene expression.

Keywords: betaine, gene expression, meat productivity, broiler chickens, immunity, inflammation, diet.

 

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