Impact of Zinc Supplementation on Tharparkar Bull Semen Quality: Comparative Study of Commercial and Lab Extenders

  • ASMATULLAH KAKA 1Department of Animal Reproduction, Faculty of Animal Husbandry and Veterinary Sciences, Sindh Agriculture University, Tandojam, 70060.
  • ABDULLAH CHANNO Pakistan Agricultural Research Council, Arid Zone Research Centre (PARC-AZRC), Umerkot.
  • ASMATULLAH KAKA Department of Animal Reproduction, Faculty of Animal Husbandry and Veterinary Sciences, Sindh Agriculture University, Tandojam
  • HABIBULLAH JANYARO Shaheed Benazir Bhutto University of Veterinary and Animal Sciences, Sakrand.
Keywords: Bull Semen


This study was aimed to evaluate the impact of zinc supplementation on chilled and frozen thawed quality parameters of Tharparkar bull semen by comparing the use of BULLXcell® (Commercial) and tris-based egg yolk (Lab based) extenders impact on semen quality parameters. The research was conducted at the Department of Animal Reproduction, Sindh Agriculture University Tandojam. Total 44 semen ejaculates were collected from four Tharparkar Bulls (A, B, C, and D) aged 4-5 years using an artificial vagina. Macroscopic and Microscopic parameters i.e. color: visual observation, volume: graduated tube, pH: digital pH meter, Progressive Motility (20x magnification), morphology and viability (eosin-nigrosin stain), concentration (haemocytometer method), and membrane integrity (HOST solution), were evaluated. Semen samples with progressive motility, morphology, viability, and membrane integrity exceeding 70% were pooled and processed. The pooled semen samples were further extended by pouring into four groups [A: BullXcell® control, B: BullXcell®+Zinc (0.2 mg/100 ml), C: Tris+Control, D: Tris+Zinc (0.2 mg/100 ml)]. Chilled assessment of progressive motility, morphology, viability, and membrane integrity revealed that Group B (78.65±0.85, 85.23±0.81, 81.22±1.17, 80.51±0.73) (P<0.05) demonstrated improved results compared to Group D (73.82±0.91, 81.66±1.14, 76.51±1.22, 76.27±1.07). Similarly, post-thawed assessment of these parameters indicated superior results for Group B (57.25±0.82, 75.17±1.02, 70.22±1.34, 63.55±1.39) (P<0.05) compared to Group D (47.35±0.66, 63.24±1.47, 65.05±1.63, 57.14±1.02) followed A and C. Based on the findings, it was concluded that Group B [BullXcell®+Zinc (0.2 mg/100 ml)] significantly improved both Chilled and post thawed quality parameters of Tharparkar cattle bull semen and the inclusion of Zinc in extenders is recommended as it improved viability of spermatozoa.


Akhter, S., Ansari, M. S., Rakha, B. A., Andrabi, S. M. H., Iqbal, S., & Ullah, N. (2010). Cryopreservation of buffalo (Bubalus bubalis) semen in Bioxcell® extender. Theriogenology, 74(6), 951-955.

Ansari, A., Aman, A., Siddiqui, N. N., & Iqbal, S. (2012). Bacteriocin (BAC-IB17): screening, isolation and production from Bacillus subtilis KIBGE IB-17. Pakistan Journal of Pharmaceutical Sciences, 25(1).

Apu, A. S., Yahia, K. M., Hussain, S. S., Fakruzzaman, M., & Notter, D. R. (2012). a comparative study of fresh and frozen‐thawed semen quality in relation to fertility of black bengal goats semen from six adult male black bengal goats. Iranian Journal of Applied Animal Sciences, 2:157-161.

Ardon, F., & Suarez, S. S. (2013). Cryopreservation increases coating of bull sperm by seminal plasma binder of sperm proteins BSP1, BSP3, and BSP5. Reproduction, 146(2), 111-117.

Channo, A., Kaka, A., Kalwar, Q., Jamali, I., Jelani, G., Bakhsh, M., Dahri, G. N., & Goil, J. P. (2022). An overview of bovine cystic ovarian disease. Pakistan Journal of Zoology, 54(5), 2437.

Channo, A., Kaka, A., Memon, A. A., Chand, M., Malhi, M. B., Kalwar, Q., Tunio, S. A., & Panhwar, M. I. (2023). Post Thawed Quality of Tharparkar Bull Semen Extended in Bioxcell TM and Tris-Based Egg Yolk Extenders supplemented with Vitamin E (α-Tocopherol). Pakistan Journal of Zoology, 1-6.

Channo, A., kaka, A., Memon, A., Malhi, M., Bakhsh, M., Kalwar, Q., Kalhoro, D., Sethar, A., & Jariko, R. (2024). Effects of α-Tocopherol on chilled quality parameters of Tharparkar bull semen extended in Lecithin and Tris based egg yolk extender. Journal of the Hellenic Veterinary Medical Society, 74(4),6511–6516.

Emamverdi, M., Zhandi, M., Shahneh, A. Z., Sharafi, M., Akhlaghi, A., Motlagh, M. K., Dadkhah, F., & Davachi, N. D. (2014). Flow cytometric and microscopic evaluation of post-thawed ram semen cryopreserved in chemically defined home-made or commercial extenders. Animal Production Science, 55(4), 551-558.

Fouad, W. F., & Ashour, A. F. (2021). Selenium and Zinc as Supplements to Extenders Frozen Semen for Improving Sperm Characteristics during Cryopreservation. Journal of Animal and Poultry Production, 12(2), 71-77.

Gürler, H., Malama, E., Heppelmann, M., Calisici, O., Leiding, C., Kastelic, J. P., & Bollwein, H. (2016). Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm. Theriogenology, 86(2), 562-571.

Kaka, A., 2015.,pp,88-118.

Kaka, A., Haron, W., Yimer, N., Channo, A., Jahejo, A. R., Ebrahimi, M., & Kalhoro, D. H. (2023). Enhancement of frozen-thawed quality of bull semen after enrichment of docosahexaenoic acid extended in tris-extender. Journal of Animal Health and Production, 11(1), 7-13.

Kaka, A., M. U. Samo, T. H. Rahoo, Z. U. Rehman, Z. Shah, M. Mushtaq, U. Kaka, and A. A. Behan. (2012). "Study on post thawing quality of kundhi buffalo semen." Journal of Animal and Plant Sciences, 22, 2: 59-62.

Kaka, A., Memon, A. A., Khatri, P., Kumbhar, H. K., Kalhoro, D. H., Tariq, M., Pirzado, S. A., & Tunio, A. N. (2016). Determination of quality characteristics of Khundi buffalo bull semen. Journal of Basic and Applied Sciences, 12, 394-397.

Kaka, A., Wahid, H., Rosnina, Y., Yimer, N., Khumran, A. M., Sarsaifi, K., Behan, A. A., & Ebrahimi, M. (2015b). α-Linolenic acid supplementation in BioXcell® extender can improve the quality of post-cooling and frozen-thawed bovine sperm. Animal reproduction science, 153, 1-7.

Kukovics, S., Gyoker, E., Németh, T., & Gergátz, E. (2011). Artificial insemination of sheep–possibilities, realities and techniques at the farm level. Artificial insemination in farm animals.

Kumar, N., Verma, R. P., Singh, L. P., Varshney, V. P., & Dass, R. S. (2006). Effect of different levels and sources of zinc supplementation on quantitative and qualitative semen attributes and serum testosterone level in crossbred cattle (Bos indicus $bftimes $ Bos taurus) bulls. Reproduction Nutrition Development, 46(6), 663-675.

Leboeuf, B., Restall, B., & Salamon, S. (2000). Production and storage of goat semen for artificial insemination. Animal reproduction science, 62(1-3), 113-141.

Memon, A. S., Kaka, A., Channo, A., Bakhsh, M., Kalwar, Q., & Sethar, A. (2022). Effect of Soybean Milk and Tris Egg Yolk Based Extender on Semen Quality Parameters of Tharparkar Bull. University of Sindh Journal of Animal Sciences (USJAS), 6(03).

Motemani, M., Chamani, M., Sharafi, M., & Masoudi, R. (2017). Alpha-tocopherol improves frozen-thawed sperm quality by reducing hydrogen peroxide during cryopreservation of bull semen. Spanish journal of agricultural research, 15(1), 15.

Najafi, A., Zhandi, M., Towhidi, A., Sharafi, M., Sharif, A. A., Motlagh, M. K., & Martinez-Pastor, F. (2013). Trehalose and glycerol have a dose-dependent synergistic effect on the post-thawing quality of ram semen cryopreserved in a soybean lecithin-based extender. Cryobiology, 66(3), 275-282.

Panhwar, M. I., Kaka, A., Memon, A. A., Channo, A., & Kolachi, H. A. (2023). In-vitro impacts of Triladyl® extender on semen quality parameters of Tharparkar bull. Pure and Applied Biology. 13(2),134-140.

Pytlík, J., Savvulidi, F. G., Ducháček, J., Codl, R., Vrhel, M., Nagy, S., & Stádník, L. (2022). Effect of extender on the quality and incubation resilience of cryopreserved Holstein bull semen. Czech Journal of Animal Science, 67(3), 75-86.

Rasul, Z., Anzar, M., Jalali, S., & Ahmad, N. (2000). Effect of buffering systems on post-thaw motion characteristics, plasma membrane integrity, and acrosome morphology of buffalo spermatozoa. Animal reproduction science, 59(1-2), 31-41.

Raval, R. J., & Dhami, A. J. (2010). Effect of additives on various spermatozoal attributes of fresh, frozen-thawed and refrigerated semen. The Indian Journal of Animal Reproduction, 31(1), 33-36.

Rehman, F., Zhao, C., Shah, M. A., Qureshi, M. S., & Wang, X. (2013). Semen extenders and artificial insemination in ruminants. Veterinaria, 1(1), 1-8.

Shah, S. H., Memon, A. A., Kaka, A., Tunio, A. N., Channo, A., Kalwar, Q., Panhwar, M. I., Malak, K. A., & Rahoo, B. A. (2023). Evaluation of Ovsynch Estrus Synchronization in Holstein Friesian and Holstein Friesian-Jersey Crossbred Cows at District Thatta, Sindh, Pakistan. Pakistan Journal of Zoology. 1-8.

Singh, M. A. H. A. K., RAMTEKE, S. S., GHOSH, S. K., Prasad, J. K., & Rajoriya, J. S. (2013). Efficacy of egg yolk from three avian species on semen freezability of Tharparkar bull. The Indian Journal of Animal Reproduction, 34(2), 25-28.

Towhidi, A., & Parks, J. E. (2012). Effect of n-3 fatty acids and α-tocopherol on post-thaw parameters and fatty acid composition of bovine sperm. Journal of assisted reproduction and genetics, 29, 1051-1056.

Yadav, D., Singh, V., Yadav, S. S., Patel, A., Kumar, A., Sonkar, V., Kumar, A., Yadav, S., Kumar, B., Yadav, B., & Saxena, A. (2019). Effect of glutathione on viability and progressive motility of Hariana bull spermatozoa during cryopreservation in semi-arid region. Journal of pharmaceutical innovations, 8(6), 8-876.