Sex ratio of calves generated from artificial insemination using frozen-thawed epididymal spermatozoa of spotted buffalo

Paper Details

Research Paper 01/02/2017
Views (423) Download (17)

Sex ratio of calves generated from artificial insemination using frozen-thawed epididymal spermatozoa of spotted buffalo

Muhammad Rizal, Yulnawati Yusnizar, Jane M. Morrell, Arief Boediono
Int. J. Biosci.10( 2), 6-11, February 2017.
Certificate: IJB 2017 [Generate Certificate]


Utilization of the cauda epididymis as an alternative spermatozoa source for assisted reproductive technology (ART) has been adopted in many species.  This technique is useful to preserve genetic diversity in endangered species, and from animals experiencing sudden death or pathology resulting in an inability to ejaculate.  This technique has been used for several decades, but data about the distribution of the sex ratio of the offspring produced is limited.  In a preliminary artificial insemination (AI) study concerning using frozen-thawed epididymal spermatozoa from the spotted buffalo, a skewed sex ratio was observed.  Results of this study showed that mean progressive motility, viability, and membrane integrity of fresh epididymal spermatozoa were 73.3%, 85.4%, and 86.2%, decreasing to 43.3%, 66.6%, and 66.9% after thawing, respectively. We got high and with a significantly higher number of female (12; 85.7%) compare to male (2; 14.3%) offspring (P<0.05).  These interesting preliminary results indicate that a further study is needed.


Akhter S, Ansari MS, Rakha BA, Andrabi SMH, Iqbal S, Ullah N. 2010. Cryopreservation of buffalo (Bubalus bubalis) semen in Bioxcell® extender. Theriogenology 74, 951-955.

Akhter S, Ansari MS, Rakha BA, Andrabi SMH, Ullah N, Khalid M. 2012. Soya-lecithin in extender improves the freezability and fertility of buffalo (Bubalus bubalis) bull spermatozoa. Reproduction in Domestic Animals 47, 815-819.

Álvarez M, Tamayo-Canul J, Martínez-Rodríguez C, López-Urueña E, Gomes-Alves S, Anel L. 2012. Specificity of the extender used for freezing ram sperm depends of the spermatozoa source (ejaculate, electroejaculate or epididymis). Animal Reproduction Science 132, 145-154.

Blash S, Melican D, Gavin W. 2000. Cryopreservation of epididymal sperm obtained at necropsy from goats. Theriogenology 5, 899-905.

Checa ML, Dunner S, Canon J. 2002. Prediction of X and Y chromosome content in bovine sperm by using DNA pools through capillary electrophoresis. Theriogenology 58, 1579-1586.

Gañán N, Gomendio M, Roldan ERS. 2009. Effect of storage of domestic cat (Felis catus) epididymides at 5 ºC on sperm quality and cryopreservation. Theriogenology 72, 1268-1277.

Gloria A, Contri A, de Amicis I, Robbe D, Carluccio A. 2011. Differences between epididymal and ejaculated sperm characteristics in donkey. Animal Reproduction Science 128, 117-122.

Guerrero CA, Gentry G, Saenz J, Bondioli KR, Godke RA. 2008. Birth of calves after artificial insemination with cryopreserved bovine epididymal spermatozoa harvested from postmortem bulls [abstract]. Reproduction, Fertility and Development 21, 105.

Heise A, Kähn W, Volkmann DH, Thompson PN, Gerber D. 2010. Influence of seminal plasma on fertility of fresh and frozen-thawed stallion epididymal spermatozoa. Animal Reproduction Science 118, 48-53.

Hermansson U, Axnér E. 2007. Epididymal and ejaculated cat spermatozoa are resistant to cold shock but egg yolk promotes sperm longevity during cold storage at 4 ºC. Theriogenology 67, 1239-1248.

Kochhar HPS, Peippo J, King WA. 2001. Sex related embryo development. Theriogenology 55, 3-14.

Kochhar HPS, Kochhar KP, Basrur PK, King WA. 2003. Influence of the duration of gamete interaction on cleavage, growth rate and sex distribution of in vitro produced bovine embryos. Animal Reproduction Science 77, 33-49.

Kozdrowski R, Niżański W, Dubiel A, Olech W. 2011. Possibilities of using the European bison (Bison bonasus) epididymal spermatozoa collected post-mortem for cryopreservation and artificial insemination: a pilot study. Reproductive Biology and Endocrinology 9, 31-36.

Lechniak D, Strabel T, Bousquet D, King AW. 2003. Sperm pre-incubation prior to insemination affects the sex ratio of bovine embryos produced in vitro. Animal Reproduction Science 38, 224-227.

Lone FA, Islam R, Khan MZ, Sofi KA. 2011. Effect of transportation temperature on the quality of cauda epididymal spermatozoa of ram. Animal Reproduction Science 123, 54-59.

Lone FA, Islam R, Khan MZ, Sofi KA. 2012. Effect of different egg yolk-based extenders on the quality of ovine cauda epididymal spermatozoa during storage at 4 °C. Reproduction in Domestic Animals 47, 257-262.

Madrid-Bury N, Fernandez R, Jimenez A, Perez-Garnelo S, Moreira PN, Pintado B, de la Fuente J. 2003. Effect of ejaculate, bull, and a double swim-up sperm processing method on sperm sex ratio. Zygote 11, 229-235.

Martinez-Pastor F, Guerra C, Kaabi M, Diaz AR, Anel E, Herraez P. 2005. Decay of sperm obtained from epididymis of wild ruminants depending on postmortem time. Theriogenology 63, 24-40.

Martins CF, Rumpf R, Pereira DC, Dode MN. 2007. Cryopreservation of epididymal bovine spermatozoa from dead animals and its uses in vitro embryo production. Animal Reproduction Science 101, 326-331.

Morris LHA. 2004. Low dose insemination in the mare: an update. Animal Reproduction Science 82, 625-632.

Papa FO, Melo CM, Fioratti EG, Dell’Aqua Jr JA, Zahn FS, Alvarenga MA. 2008. Freezing of stallion epididymal sperm. Animal Reproduction Science 107, 293-301.

Parati K, Bongioni G, Aleandri R, Galli A. 2006. Sex ratio determination in bovine semen: A new approach by quantitative real time PCR. Theriogenology 66, 2202-2209.

Rasul Z, Ahmad N, Anzar M. 2001. Changes in motion characteristics, plasma membrane integrity and acrosome morphology during cryopreservation of buffalo spermatozoa. Journal of Andrology 22, 278-283.

Revell SG, Mrode RA. 1994. An osmotic resistance test for bovine semen. Animal Reproduction Science 36, 77-86.

Saenz JR. 2007. Cryopreservation of white-tail deer epididymal sperm for artificial insemination. PhD Thesis, Louisiana State University, United State of America.

Santiago-Moreno J, Toledano-Díaz A, Pulido-Pastor A, Gómez-Brunet A, López-Sebastián A. 2006. Birth of live Spanish ibex (Capra pyrenaica hispanica) derived from artificial insemination with epididymal spermatozoa retrieved after death. Theriogenology 66, 283-291.

Santiago-Moreno J, Coloma MA, Toledano-Díaz A, Gómez-Brunet A, Pulido-Pastor A, Zamora-Soria A. 2008. A comparison of the protective action of chicken and quail egg yolk in the cryopreservation of Spanish ibex epididymal spermatozoa. Cryobiology 57, 25-29.

Singh AK, Singh VK, Narwade BM, Mohanty TK, Atreja SK. 2012. Comparative quality assessment of buffalo (Bubalus bubalis) semen chilled (5 °C) in egg yolk- and soya milk-based extenders. Reproduction in Domestic Animals 47, 596-600.

Soler AJ, García AJ, Fernández-Santos MR, Esteso MC, Garde JJ. 2003. Effects of thawing procedure on post-thawed in vitro viability and in vivo fertility of red deer epididymal spermatozoa cryopreserved at -190 °C. Journal of Andrology 24, 746-56.

Turri F, Madeddu M, Gliozzi TM, Gandini G, Pizzi F. 2012. Influence of recovery methods and extenders on bull epididymal spermatozoa quality. Reproduction in Domestic Animals 47, 712-717.

Waheed MM, Al-Eknah MM, El-Bahr SM. 2011. Some biochemical characteristics and preservation of epididymal camel spermatozoa (Camelus dromedarius). Theriogenology 76, 1126-1133.

Waheed S, Ahmad N, Najib-ur-Rahman, Jamil-ur-Rahman H, Younis M, Iqbal S. 2012. Evaluation of duck egg yolk for the cryopreservation of Nili-Ravi buffalo bull semen. Animal Reproduction Science 131, 95-99.