Cryoprotective Effects of Natural Honey Supplementation on Post-Thaw Sperm Quality of Nili-Ravi Buffalo (Bubalus bubalis)

Mishal Rubab; Aasma Iqbal

doi:10.61504/

Authors

Mishal Rubab Department of Zoology, University of Lahore, Sargodha Campus, Sargodha, Pakistan Author
Aasma Iqbal Department of Zoology, University of Lahore, Sargodha Campus, Sargodha, Pakistan Author

DOI:

https://doi.org/10.61504/

Keywords:

Cryoprotective, Natural Honey, Sperm Quality, Nili-Ravi Buffalo

Abstract

Cryopreservation is essential for buffalo breeding and artificial
insemination (AI) programs. Freezing-thawing process often induces
cryo-damage, compromising sperm quality. Conventional
cryoprotectants can also exert toxic effects, creating the need for safer,
natural alternatives. This study evaluated the cryoprotective potential
of natural honey supplementation in a Tris–Fructose–Egg-Yolk (TFEY)
extender for Nili-Ravi buffalo (Bubalus bubalis) spermatozoa. Semen
from healthy Nili-Ravi bulls was diluted with TFEY extender containing
0% (control), 2.5%, 5.0%, 7.5%, or 10% (v/v) natural honey, equilibrated,
and cryopreserved in 0.5 mL straws. Sperm quality parameters:
progressive motility, plasma membrane integrity (PMI; via hypo
osmotic swelling test), and livability were evaluated at both post
dilution and post-thaw stages. Data were analyzed using one-way
ANOVA followed by Tukey’s HSD test (p < 0.05. The findings
demonstrate honey’s potential as a natural and biocompatible
cryoprotectant, offering a promising approach to improve AI success
rates and support sustainable buffalo production.