The Post Thaw Quality, Antioxidant Activity, and In vivo Fertility of Frozen-Thawed Buffalo Bull Semen Frozen-Stored in the Presence of Different Concentrations of Freeze-Dried Clove Powder

Document Type : Original Article

Authors

1 El-Geish Street

2 Department of Theriogenology, Faculty of Veterinary Medicine, Kafrelsheikh University, Egypt

3 Biotechnology Research Department, Animal Production Research Institute, Agriculture Research Centre, Giza, Egypt.

4 Department of Biotechnology, Animal Production Research Institute, Egypt

5 Kafrelsheikh University, Faculty of Veterinary Medicine, Department of Theriogenology, El-Geish Street, Kafrelsheikh 33516, Egypt.

6 Department of Theriogenology, Faculty of Veterinary Medicine, Kafrelsheikh University, Egypt.

Abstract

Artificial insemination (AI) is a critical reproductive technology in the buffalo industry, offering numerous benefits for improving genetic quality by selective breeding and faster genetic progress. The plasma membrane of buffalo bull spermatozoa is rich in polyunsaturated fatty acids and is susceptible to lipid peroxidation during freezing and thawing processes. The addition of herbal extracts to buffalo bull semen extenders has been explored as a potential method to improve semen quality and fertility. The most potent antioxidants in clove bud extract are phenolic compounds, particularly eugenol, which is the major active component. This study aims to minimize the damaging effect of oxidative stress during freezing-thawing procedures, in turn improving the post-thaw quality and in vivo fertility of buffalo bull spermatozoa. Ejaculates were collected from seven healthy Egyptian buffalo bull by artificial vagina twice weekly. Good-quality ejaculates were pooled and dispensed into 4 Aliquots; Each aliquot was diluted with Tris-egg yolk citric acid diluent containing different concentrations of clove buds (Syzygium aromaticum) extract as follows: 1) 0 μg/mL as a control (CTRL), 2) 5 μg/mL (CLOV5), 3) 10 μg/mL (CLOV10), and 4) 15 μg/mL (CLOV15). Diluted semen samples were equilibrated at 4 °C for 4 hours, loaded into French mini straws, sealed, frozen stored in liquid nitrogen. Frozen straws were thawed and examined for sperm characteristics; also, enzymatic antioxidants (SOD, CAT, GPX) as well as the lipid peroxidation marker (MDA) were determined. A total of 84 (21 females/group) mature and healthy buffalo-cows were selected, exposed to estrus synchronization, and inseminated by the prepared frozen-thawed straws to calculate their in vivo fertility rates. The results revealed that CLOV15 and CLOV10 have higher (P < 0.05) proportions of all post-thaw sperm characteristics, and also, CLOV15 yielded the greatest pregnancy rate, 66.66%, in comparison with CLOV5 (33.33%) and CLOV10 (11.11%). Niether SOD nor CAT affected by the clove addition whereas the GPX activity improved in CLOV10 compared with other group. In the same way, MDA content was lower in the higher concentration of clove compared with the low concentration and control groups. In conclusion, enriching cryopreservation extender with 15µg/ml clove buds powder improved almost all sperm characteristics. On the other side, CLOV10 and CLOV5 improved the enzymatic antioxidant, and only CLOV15 improved the in vivo fertility rate of frozen-thawed buffalo bull semen.

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