Successful Separation of X- and Y-Spermatozoa Ongole Crossbreed Using a Nano-Albumen Gradient Column
Abstract
The objective of this study was to determine the efficacy of separating X- and Y-sperm using nano-albumen (NA). The material used in this study was fresh semen of Ongole Crossbreed bulls with progressive motility of ≥ 70%. Treatments for the top and bottom fractions were T0 (BSA 5% : 10%) as a control, T1 (NA 2% : 4%), T2 (NA 4% : 6%), and T3 (NA 6% : 8%). Sexed semen was evaluated for characteristics and kinematics, and the proportions of sexed semen were validated using morphometric and polymerase chain reaction methods. The data on the characteristics and motility kinematics of X- and Y-sperm were analyzed by ANOVA using Minitab 18. The results revealed that the semen separated by the NA column generally showed the same sperm quality characteristics as the BSA column. However, the total motility of the T3y treatment in the bottom fraction was significantly lower (p<0.05) compared to the control. The kinematics of spermatozoa motility in the top fraction was not significantly different; however, the progressive motility value of T3y was significantly different (p<0.05) compared to that of T1y, and the BCF value of T2y was significantly different from that of the control in the bottom fraction. The acrosome status in the top fraction was better than in the bottom fraction, and the DNA integrity value of the top and bottom fractions showed DNA damage ranging from 2% to 5%. The present study indicated that: (1) the Nano-Albumen gradient column could be used successfully to separate X- and Y-spermatozoa, (2) characteristic values and sperm kinematics, acrosomal status, and DNA integrity were comparable to those of the BSA control, (3) combination column in treatments NA 4% (T2x) and 6% (T2y) appeared the good ability to separated X and Y-sperm.
References
Amal, A. S., R. I. Arifiantini, M. A. Setiadi, & S. Said. 2019. Characteristics of the post-thawed Balinese bull semen extended in three different extenders and equilibration times. J. Indones. Trop. Anim. Agric. 44:135-145. https://doi.org/10.14710/jitaa.44.2.135-145
Arif, A. A., T. Maulana, E. M. Kaiin, B. Purwantara, R. I. Arifiantini, & E. Memili. 2020. Comparative analysis of various step-dilution techniques on the quality of frozen Limousin bull semen. Vet. World. 13:2422-2428. https://doi.org/10.14202/vetworld.2020.2422-2428
Arishima, T., S. Sasaki, T. Isobe, Y. Ikebata, S. Shimbara, S. Ikeda, K. Kawashima, Y. Suzuki, M. Watanabe, S. Sugano, K. Mizoshita, & Y. Sugimoto. 2017. A maternal variant in the upstream of FOXP3 gene on the X chromosome is associated with recurrent infertility in Japanese Black cattle. BMC Genet. 18:103. https://doi.org/10.1186/s12863-017-0573-8
Blondin, P., M. Beaulieu, V. Fournier, N. Morin, L. Crawford, P. Madan, & W. A. King. 2009. Analysis of bovine sexed sperm for IVF from sorting to the embryo. Theriogenology 71:30-38. https://doi.org/10.1016/j.theriogenology.2008.09.017
[BSN] Badan Standarisasi Nasional. 2017. Semen Beku-Bagian 1: Sapi. SNI 4869-1. BSN, Jakarta (ID).
Burroughs, C. A., J. K. Graham, R. W. Lenz, & G. E. Seidel Jr. 2013. Seminal plasma effects on sex sorting bovine sperm. Theriogenology. 79:551-557. https://doi.org/10.1016/j.theriogenology.2012.10.024
Costa, M. Z., L. Z. Oliveira, M. V. Resende, A. C. Lucio, A. P. Perini, M. C. V. Miguel, & V. F. M. H. Lima. 2010. Induction of the acrosome reaction test to in vitro estimate embryo production in Nelore Cattle. Arq. Bras. Med. Vet. Zootec. 62:771-777. https://doi.org/10.1590/S0102-09352010000400001
De Canio, M., A. Soggiu, C. Piras, L. Bonizzi, A. Galli, A. Urbani, & P. Roncada. 2014. Differential protein profile in sexed bovine semen: Shotgun proteomics investigation. Mol. Biosyst. 10:1264-1271. https://doi.org/10.1039/C3MB70306A
Dwiastuti, R., S. Noegrohati, E. P. Istyastono, & Marchaban. 2016. Metode pemanasan dan sonikasi menghasilkan nanoliposom dari fosfolipid lesitin kedelai (soy lecithin). Jurnal Farmasi Sains dan Komunitas (Journal of Pharmaceutical Sciences and Community) 13:23-37. https://doi.org/10.24071/jpsc.131126
Garner, D. L. 2006. Flowcytometric sexing of mammalian sperm. Theriogenology 65:943-957. https://doi.org/10.1016/j.theriogenology.2005.09.009
Hafez, E. S. E. & B. Hafez. 2000. Reproduction in Farm Animals. 7 Edition. Lippincott Williams & Wilkins, Baltimore. https://doi.org/10.1002/9781119265306
Insani, K., R. Sri, P. Agung, & S. Aries. 2014. Kadar MDA spermatozoa setelah pembekuan. Biotropika 2:142-147.
Kaiin, E. M. & M. Gunawan. 2018. Kualitas sperma sapi hasil sexing setelah kapasitasi secara in vitro. Prosiding Seminar Nasional Masyarakat Biodiversitas Indonesia 3:466-470.
Kaiin, E. M., M. Gunawan, & F. Afiati. 2013. Production of Frozen Sexing Sperm Separated with BSA Column Method with Standardization on Artificial Insemination Center. International Conference on Biotechnology, Bogor. pp. 67-72.
Kathiravan, P., J. Kalatharan, G. Karthikeya, K. Rengarajan, & G. Kadirve. 2011. Objective sperm motion analysis to assess dairy bull fertility using computer-aided system. A review. Reprod. Domest. Anim. 46:165-172. https://doi.org/10.1111/j.1439-0531.2010.01603.x
Kusumawati E. D., N. Isnaini, A. P. A. Yekti, M. Luthfi, L. Affandhy, D. Pamungkas, Kuswati, A. Ridhowi, H. Sudarwati, T. Susilawati, & S. Rahayu. 2017. The quality of sexed semen on filial ongole bull using percoll density gradient centrifugation method. Asian J. Microbiol. Biotechol Environ. Sci. 19:189-199.
Malik, A., W. Haron, R. Yusoff, M. Bukar, A. Kasim, & M. Sabri. 2011. Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility, and membrane integrity in bovine. Acad. J. Biotechnol. 10:19796-19801. https://doi.org/10.5897/AJB11.1563
Mansur, M. A. A., M. G. S. Alam, P. K. Jha, M. A. Rimon, N. Naher & F. Y. Bari. 2018. Conception rate following intra-cervical artificial insemination using frozen semen at field level in indigenous sheep of Bangladesh. Asian J. Med. Biol. Res. 4:55–62. https://doi.org/10.3329/ajmbr.v4i1.36822
Maulana, T., S. Said, R. I. Arifiantini, & M. A. Setiadi. 2019. Sex sorting sperm of Sumba Ongole bulls by using snakehead fish (Channa striata) albumin extract. J. Indones. Trop. Anim. Agric. 44:106-113. https://doi.org/10.14710/jitaa.44.1.106-113
Miranda, P. V., A. Allaire, J. Sosnik, & P. E. Visconti. 2009. Localization of low-density detergent-resistant membrane proteins in intact and acrosome-reacted mouse sperm. Biol. Reprod. 80:897-904. https://doi.org/10.1095/biolreprod.108.075242
Nagy, A., T. Polichronopoulos, A. Gaspardy, L. Solti, & S. Cseh. 2015. Correlation between bull fertility and sperm cell velocity parameters generated by computer-assisted semen analysis. Acta Vet. Hung. 63:370–381. https://doi.org/10.1556/004.2015.035
Oliveira, L. Z., R. P. de Arruda, A. F. C. de Andrade, E. C. C. Celeghini, P. D. Reeb, & V. F. M. H. de Lima. 2013. Assessment of in vitro sperm characteristics and their importance in the prediction of conception rate in a bovine timed-AI program. Anim. Reprod. Sci. 137:145-155. https://doi.org/10.1016/j.anireprosci.2013.01.010
Paddrik, P., T. Hallap, T. Kaart, T. Bulitko, & U. Jaakmal. 2012. Relationships between the results of hypoosmotic swelling tests, sperm motility, and fertility in Estonian Holstein dairy bulls. Czech. J. Anim. Sci. 57:490-497. https://doi.org/10.17221/6349-CJAS
Penfold, L. M., C. Holt, W. V. Holt, G. R. Welch, D. G. Cran, & L. A. Johnson. 1998. Comparative motility of X and Y chromosome bearing bovine sperm separated on the basis of DNA content by flow sorting. Mol. Reprod. Dev. 50:323-32. https://doi.org/10.1002/(SICI)1098-2795(199807)50:3<323::AID-MRD8>3.0.CO;2-L
Perumal, P., S. K. Srivastava, S. K. Ghosh, & K. K. Baruah. 2014. Computer assisted sperm analysis of freezable and nonfreezable mithun (Bis frontalis) semen. J. Anim. 1:1-6. https://doi.org/10.1155/2014/675031
Prakash, M. A., A. Kumaresan, A. Manimaran, R. K. Joshi, S. S. Layek, T. K. Mohanty, & R. R. Divisha. 2014. Sexing of spermatozoa in farm animals: A mini-review. Adv. Anim. Vet. Sci. 2:226-232. https://doi.org/10.14737/journal.aavs/2014/2.4.226.232
Rajabi-Toustani, R., Q. S. Akter, E. A. Alamadaly, Y. Hoshino, H. Adachi, K. Mukoujima, & T. Murase. 2019. Methodological improvement of fluorescein isothiocyanate peanut agglutinin (FITC-PNA) acrosomal integrity staining for frozen-thawed Japanese Black bull spermatozoa. J. Vet. Med. Sci. 81:694-702. https://doi.org/10.1292/jvms.18-0560
Rattanasuk, S., R. Parnpai, & M. Ketudat-Cairns. 2011. Multiplex polymerase chain reaction used for bovine embryo sex determination. J. Reprod. Dev. 57:539-542. https://doi.org/10.1262/jrd.10-126M
Said, S., F. Afiati, & T. Maulana. 2015. Study on changes of sperm head morphometry and DNA integrity of freeze-dried bovine spermatozoa. J. Indones. Trop. Anim. Agric. 40:145-152. https://doi.org/10.14710/jitaa.40.3.145-152
Santolaria, P., A. Pauciullo, M. A. Silvestre, S. Vicente-Fiel, L. Villanova, A. Pinton, J. Viruel, E. Sales, & J. L. Yániz. 2016. Computer-assisted sperm morphometry fluorescence-based analysis has potential to determine progeny sex. Asian J. Androl. 18:858-862. https://doi.org/10.4103/1008-682X.187578
Shojaei, H., T. Kroetsch, R. Wilde, P. Blondin, J. Kastelic, & J. C. Thundathil. 2012. Moribund sperm in frozen-thawed semen, and sperm motion end points post-thaw and post-swim-up, are related to fertility in Holstein AI bulls. Theriogenology 77:940-951. https://doi.org/10.1016/j.theriogenology.2011.09.026
Sarastina, T. Susilawati, & G. Ciptadi. 2007. Analisa beberapa parameter motilitas spermatozoa pada berbagai bangsa sapi menggunakan computer assisted semen analysis (CASA). Ternak Tropika (Journal of Tropical Animal Production) 6:1-12.
Sharma, M. & N. Sharma. 2016. Sperm sexing in animals. Adv. Anim. Vet. Sci. 4:543-549. https://doi.org/10.14737/journal.aavs/2016/4.10.543.549
Susilawati, T. 2011. Spermatology. UB Press, Malang.
Susilawati, T., Kusumawati, D. Ratnawati, N. Isnaini, & A. P. Yekti. 2018. Character of liquid semen motility in various diluents on balinese cattle during cold storage. Asian J. Microbiol. Biotechnol. Environ. Sci. 20:166-172.
Yang, W. C., L. Sang, Y. Xiao, H. L. Zhang, K. Q. Tang, & L. G. Yang. 2014. Tentative identification of sex-specific antibodies and their application for screening bovine sperm proteins for sex-specificity. Mol. Biol. Rep. 41:217-223. https://doi.org/10.1007/s11033-013-2854-2
Yániz, J. L., C. Soler, & P. Santolaria. 2015. Computer assisted sperm morphometry in mammals: A review. Anim. Reprod. Sci. 156:1–12. https://doi.org/10.1016/j.anireprosci.2015.03.002
Authors
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Authors submitting manuscripts should understand and agree that copyright of manuscripts of the article shall be assigned/transferred to Tropical Animal Science Journal. The statement to release the copyright to Tropical Animal Science Journal is stated in Form A. This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License (CC BY-SA) where Authors and Readers can copy and redistribute the material in any medium or format, as well as remix, transform, and build upon the material for any purpose, but they must give appropriate credit (cite to the article or content), provide a link to the license, and indicate if changes were made. If you remix, transform, or build upon the material, you must distribute your contributions under the same license as the original.
Article Details
