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Birth of the first mithun (Bos frontalis) calf through artificial insemination

  • A. Dhali (a1), M. Karunakaran (a2), A. Mech (a1), N. Nath (a1), B. Prakash (a1), C. Rajkhowa (a1) and D. P. Mishra (a1)...


The study describes the standardization of a suitable semen cryopreservation protocol for the first time in mithun (Bos frontalis) and birth of the first mithun calf through artificial insemination. The semen samples were collected from adult bulls through the rectal massage method and cryopreserved in liquid nitrogen using tris–egg yolk–glycerol diluent. The diluted semen samples were packaged in 0.50 ml straws and kept at 5°C for 4 h for equilibration. Following the equilibration, the straws were frozen into liquid nitrogen vapour for 10 min and then plunged into liquid nitrogen for storage. It was observed that the progressive motility (%) decreased significantly (P < 0.01) in cryopreserved semen (43.3 ± 4.1) compared with fresh samples (76.6 ± 3.3). The percentages of live spermatozoa (P < 0.01) and spermatozoa with intact acrosome (P < 0.05) also decreased significantly in cryopreserved semen (54.0 ± 3.3 and 64.6 ± 5.3) compared with fresh samples (79.3 ± 2.6 and 85.3 ± 1.8). Simultaneously, the total morphological abnormality (%) was found to be significantly (P < 0.01) higher in cryopreserved samples (15.46 ± 2.68) than in fresh semen (3.85 ± 0.63). A total of three mithun cows were inseminated using the cryopreserved semen. All the cows conceived following insemination and gave birth to healthy calves. The study revealed that mithun semen can be cryopreserved efficiently using tris–egg yolk–glycerol diluent, which can be further used for artificial insemination.


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Bhattacharya, HK, Goswami, BK, Bujarbaruah, KM, Deka, BC, Baishya, N, Sarma, BC 2005. Characteristics of semen collected by massage method in mithun (Bos frontalis) bulls. Indian Journal of Animal Science 75, 11681169.
Dee Leeuw, FE, Dee Leeuw, AM, Dean Daas, JHG, Colenbrander, B, Verkeij, AJ 1993. Effect of various cryoprotective agents and membrane stabilizing compounds on bull sperm membrane integrity after cooling and freezing. Cryobiology 30, 3244.
Fennessy, PF, Mackintosh, CG, Shackell, GH 1990. Artificial insemination of farmed red deer (Cervus elaphus). Animal Production 51, 613621.
Foote, RH, Bratton, RW 1950. The fertility of bovine semen in extenders containing sulfanilamide, penicillin, streptomycin, and polymyxin. Journal of Dairy Science 33, 544547.
Foote, RH, Gray, LC, Young, DC, Dunn, HO 1960. Fertility of bull semen stored up to four days at 5°C in 20% egg yolk extenders. Journal of Dairy Science 43, 13301334.
Garland, P 1989. Artificial insemination of scimitar-horned oryx (Oryx dammah). Bulletin of Zoo Management 27, 2930.
Hammer, CJ, Tyler, HD, Loskutoff, NM, Armstrong, DL, Funk, DJ, Lindsey, BR, Simmons, LG 2001. Compromised development of calves (Bos gaurus) derived from in vitro-generated embryos and transferred interspecifically into domestic cattle (Bos taurus). Theriogenology 55, 14471455.
Holt, WV 2000. Fundamental aspects of sperm cryobiology: the importance of species and individual differences. Theriogenology 53, 4758.
Hopkins, SM, Armstrong, DL, Hummel, S, Junior, S 1988. Successful cryopreservation of gaur (Bos gaurus) epididymal spermatozoa. Journal of Zoo Animal Medicine 19, 195201.
Januskauskas, A, Lukoseviciute, K, Nagy, S, Johannisson, A, Rodriguez-martinez, H 2005. Assessment of the efficacy of Sephadex G-15 filtration of bovine spermatozoa for cryopreservation. Theriogenology 63, 160178.
Johnston, LA, Parrish, JJ, Monson, R, Leibfried-rutledge, L, Susko-parrish, JL, Northey, DL, Rutledge, JJ, Simmons, LG 1994. Oocyte maturation, fertilization and embryo development in vitro and in vivo in the gaur (Bos gaurus). Journal of Reproduction and Fertility 100, 131136.
Karunakaran, M, Dhali, A, Mech, A, Khate, K, Rajkhowa, C, Mishra, DP 2006. Preservation of mithun (Bos frontalis) semen at refrigeration temperature. Animal Reproduction Science 101, 257264.
Kovacs, A, Foote, RH 1992. Viability and acrosome staining of bull, boar and rabbit spermatozoa. Biotechnic and histochemistry 67, 119124.
Mondal, SK, Pal, DT 1999. Mithun: historical perspective. Asian Agri-History 3, 245260.
Nagy, S, Hallap, T, Johannisson, A, Rodriguez-martinez, H 2004. Changes in plasma membrane and acrosome integrity of frozen thawed bovine spermatozoa during a 4 h incubation as measured by multicolor flow cytometry. Animal Reproduction Science 80, 225235.
Salisbury, GW, Fuller, HK, Willett, EL 1941. Preservation of bovine spermatozoa in yolk-citrate diluent and field results from its use. Journal of Dairy Science 24, 905910.
Simoons, FJ 1984. Gayal or mithun. In Evolution of domesticated animals (ed. IL Manson), pp. 3436. Longman, London.
Somfai, T, Bodo, S, Nagy, S, Papp, AB, Ivancsics, J, Baranyai, B, Gocza, E, Kovacs, A 2002. Effect of swim up and Percoll treatment on viability and acrosome integrity of frozen-thawed bull spermatozoa. Reproduction in Domestic Animal 37, 285290.
SPSS 1999. SPSS® user’s guide: release 10.0.1 ed. SPSS Inc., Chicago, IL, USA.
Sukwongs Y, Kamolnorranath S, Lohachit C, Apimeteetumrong M, Yiengvisavakul V and Wolfe B 1998. Successful cryopreservation of gaur (Bibos gaurus) and banteng (Bibos javanicus) spermatozoa and pregnancies after artificial insemination with frozen semen. Proceedings of 36th Veterinary Conference, Kasetsart University, Bangkok, 8pp.
Thun, A, Hurtado, M, Janett, F 2002. Comparison of biociphos-plus® and tris-egg yolk extender for cryopreservation of bull semen. Theriogenology 57, 10871094.
Vyas, S, Mohan, G, Dhami, AJ, Sahni, KL 1992. Effect of filtration through sephadex and glasswool on the quality and freezability of semen of crossbred bulls. Indian Journal of Animal Science 61, 341343.


Birth of the first mithun (Bos frontalis) calf through artificial insemination

  • A. Dhali (a1), M. Karunakaran (a2), A. Mech (a1), N. Nath (a1), B. Prakash (a1), C. Rajkhowa (a1) and D. P. Mishra (a1)...


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