Supplementation of Baicalin (BC) in extender improves structural and functional characteristics, total antioxidant capacity, and in vivo fertility of ram semen

Anim Reprod Sci. 2026 May:288:108139. doi: 10.1016/j.anireprosci.2026.108139. Epub 2026 Feb 9.

Abstract

This investigation was aimed to evaluate the effects of Baicalin (BC) supplementation on ram semen quality during cooling, post-thaw sperm characteristics, oxidative status, and fertility outcomes. Semen samples were collected twice weekly from four reproductively sound rams using an artificial vagina, yielding a total of 64 ejaculates. The ejaculates were pooled and diluted at a ratio of 1:4 with a Tris-citric acid-fructose-yolk extender (TCFY), enhanced with varying concentrations of BC (0.50, 0.75, or 1 μM). Diluted, samples were loaded into 0.25 mL straws, gradually cooled to 5°C over 90 min, cryopreserved in liquid nitrogen vapor, and thawed after 24 h in a 37 °C water bath for 30 s. Results showed that BC supplementation at 0.50 and 0.75 μM significantly improved sperm quality parameters after cooling and freezing-thawing (P < 0.01), whereas 1 μM BC did not differ from the control. The highest values for progressive motility (43.36 ± 1.54 %), viability (56.14 ± 1.44 %), and membrane integrity (52.34 ± 1.22 %) were observed at 0.75 μM BC. Fertility rates following artificial insemination increased from 20.68 % in the control group to 51.66 % and 52.38 % in the 0.50 and 0.75 μM BC groups, respectively. In conclusion, supplementation of semen extenders with 0.75 μM Baicalin effectively enhances post-thaw sperm quality and fertility in rams.

Keywords: Baicalin; Extender; Fertility; Ram; Reproduction.

MeSH terms

  • Animals
  • Antioxidants* / pharmacology
  • Cryopreservation / veterinary
  • Cryoprotective Agents* / pharmacology
  • Female
  • Fertility* / drug effects
  • Flavonoids* / administration & dosage
  • Flavonoids* / pharmacology
  • Insemination, Artificial / veterinary
  • Male
  • Semen Analysis / veterinary
  • Semen Preservation* / methods
  • Semen Preservation* / veterinary
  • Semen* / drug effects
  • Semen* / physiology
  • Sheep / physiology
  • Sperm Motility / drug effects
  • Spermatozoa / drug effects
  • Spermatozoa / physiology

Substances

  • baicalin
  • Flavonoids
  • Antioxidants
  • Cryoprotective Agents