Improving indigenous Vietnamese Black Rabbit frozen sperm quality: the role of glycine and sperm selection methods

Authors

DOI:

https://doi.org/10.4995/wrs.2023.19690

Keywords:

cryopreservation, glycine, indigenous black rabbit, rabbit sperm, swim-up

Abstract

Rabbit sperm are known to undergo damage during both cryopreservation and thawing, leading to decreased viability, motility and membrane integrity. Glycine can protect sperm and reduce damage during freezing. Swim-up is a simple semen processing method for selecting good motile sperm. The study evaluated the effect of the swim-up method and glycine with different concentrations supplemented to the frozen medium. Three indigenous black rabbits were selected for semen collection by artificial vagina. Next, semen was selected by swim-up method and diluted with glycine-added frozen medium. The samples were then transferred to 0.5 mL straws, cooled to 15°C and 5°C, placed in liquid nitrogen vapour, and finally placed directly into liquid nitrogen (-196°C). The samples were thawed and evaluated for sperm quality. The results showed that the medium supplemented with 10mM glycine in combination with swim-up method for 30 min gave the best results and was significantly different from the remaining concentrations (P<0.01), with viability rate, overall mobility and membrane integrity of 68.0%, 58.7% and 49.7%, respectively. In conclusion, 10 mM glycine concentration combined with swim-up for 30 min is the optimal choice for freezing local black rabbit semen. The study highlights the importance of optimising freezing protocols to improve the quality of frozen rabbit sperm, which can have important implications for animal breeding and conservation efforts.

Downloads

Download data is not yet available.

Author Biographies

Tran Thi Thanh Khuong, Can Tho University

Stem cell Laboratory, Institute of Food and Biotechnology

Nguyen Lam Khanh Duy, Can Tho University

Stem cell Laboratory, Institute of Food and Biotechnology

Nguyen Thuy Hang, Can Tho University

Stem cell Laboratory, Institute of Food and Biotechnology

Phan Kim Ngoc, Ho Chi Minh City University of Science

Faculty of Biology and Biotechnology

Duong Nguyen Duy Tuyen, Phuong Chi Hospital

IVF My Duc Unit

References

Agha-Rahimi A., Khalili M., Nabi A., Ashourzadeh S. 2014. Vitrification is not superior to rapid freezing of normozoospermic spermatozoa: effects on sperm parameters, DNA fragmentation and hyaluronan binding. BioMedicine Online, 28: 352-58. https://doi.org/10.1016/j.rbmo.2013.11.015

Aitken R.J., Drevet J.R., Moazamian A., Gharagozloo P. 2022. Male Infertility and Oxidative Stress: A Focus on the Underlying Mechanisms. Antioxidants (Basel), 11: 306. https://doi.org/10.3390/antiox11020306

Alonge S., Melandri M., Leoci R., Lacalandra G.M., Caira M., Aiudi, G.G. 2019. The effect of dietary supplementation of vitamin e, selenium, zinc, folic acid, and N-3 polyunsaturated fatty acids on sperm motility and membrane properties in dogs. Animals, 9: 34. https://doi.org/10.3390/ani9020034

Alvarez J.G., Storey B.T. 1992. Evidence for increased lipid peroxidative damage and loss of superoxide dismutase activity as a mode of sublethal cryodamage to human sperm during cryopreservation. J. Androl. 13: 232-241. https://doi.org/10.1002/j.1939-4640.1992.tb00306.x

Bansal A.K., Bilaspuri G.S. 2010. Impacts of oxidative stress and antioxidants on semen functions. Vet. Med. Int., 2010: 686137. https://doi.org/10.4061/2011/686137

Butt F., Chohan M.A. 2016. Comparative efficacy of density gradient and swim-up methods of semen preparation in intrauterine insemination cycles. J. Paki. Med. Assoc., 66: 932-937.

Bucak M.N., Sariözkan S., Tuncer P.B. et al. 2010. The effect of antioxidants on post-thawed Angora goat (Capra hircus ancryrensis) sperm parameters, lipid peroxidation and antioxidant activities. Small Ruminant Res., 89: 24-30. https://doi.org/10.1016/j.smallrumres.2009.11.015

Buranaamnuay K. 2020. Effect of different permeable cryoprotectants on the quality of cat epididymal spermatozoa. Cryo Letters, 41: 237-244.

Chaveiro A., Santos P., Da Silva F. 2007. Assessment of sperm apoptosis in cryopreserved bull semen after swim-up treatment: A flow cytometric study. Reprod. Domest. Anim., 42: 17-21. https://doi.org/10.1111/j.1439-0531.2006.00712.x

Contreras M.J., Treulen F., Arias M.E. 2020. Cryopreservation of stallion semen: effect of adding antioxidants to the freezing extender on sperm physiology. Reprod. Domes. Anim., 55: 229-239. https://doi.org/10.1111/rda.13611

Di Santo, M., Tarozzi, N., Nadalini, M., Borini, A. 2012. Human sperm cryopreservation: update on techniques, effect on DNA Integrity, and Implications for ART. Adv. Urol., 2012: 854837. https://doi.org/10.1155/2012/854837

Dorado J., Gálvez M.J., Demyda-Peyrás S., Ortiz I., Morrell J.M., Crespo F., Gosálvez J., Hidalgo M. 2016. Differences in preservation of canine chilled semen using simple sperm washing, single-layer centrifugation and modified swim-up preparation techniques. Reprod. Fertil. Dev., 28: 1545-1552. https://doi.org/10.1071/RD15071

El-Sheshtawy R.I., El-Sisy G.A., El-Nattat W.S. 2008. Use of selected amino acids to improve buffalo bull semen cryopreservation. Global Vet., 2: 146-150.

Fumuso F.G., Giulianob S.M., Chavesa M.G., Neilda D.M., Miragayaa M.H., Gambarottac M.C., Carreteroa, M.I. 2018. Seminal plasma affects the survival rate and motility pattern of raw llama spermatozoa. Anim. Reprod. Sci., 192: 99-106. https://doi.org/10.1016/j.anireprosci.2018.02.019

Gadea J., Molla M., Selles E., Marco M. A., Garcia-Vazquez F.A., Gardon J.C. 2011. Reduced glutathione content in human sperm is decreased after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders. Cryobiology, 62: 40-46. https://doi.org/10.1016/j.cryobiol.2010.12.001

Henkel R.R., Schill W.B. 2013. Sperm preparation for ART. Reprod Biol Endocrinol, 1: 108. https://doi.org/10.1186/1477-7827-1-108

Hoogewijs M., Piepers S., Govaere J., Schauwer C., Kruif A., Morrell J.M. 2012. Sperm longevity following prefreeze sperm selection. J. Equine Vet.y Sci., 32: 489. https://doi.org/10.1016/j.jevs.2012.06.048

Husna A., Awan M.A., Mehmood A., Sultana T., Shahzad Q., Ansari M.S., Rakha B.A., Naqvi S.M.S., Akhter S. 2016. Sperm sexing in Nili-Ravi buffalo through modified swim up: Validation using SYBR® green real-time PCR. Anim. Reprod. Sci., 182: 69-76. https://doi.org/10.1016/j.anireprosci.2017.04.011

Jameel T. 2008. Sperm swim-up: a simple and effective technique of semen processing for intrauterine insemination. The J. Pak. Med. Assoc., 58: 71-4. Corpus ID: 29494796.

Karabekir S.C., Özgörgülü, A. 2019. Evaluation of semen samples before and after ‘Swim Up’ technique with Mitotracker. Bangladesh J. Medi. Sci., 18: 479-483. https://doi.org/10.3329/bjms.v18i3.41614.

Khalili B., Jafaroghli M., Farshad A., Paresh-Khiavi M. 2010. The effects of different concentrations of glycine and cysteine on the freezability of Moghani ram spermatozoa. Asian Austral. J. Anim. Sci., 23: 318-325. https://doi.org/10.5713/ajas.2010.90387

Khanam S., Shati M.F., Shuvro M.A., Ahmed S., Obayed T., Khanum T. 2018. Changes of quality of semen after Swim up method during intrauterine insemination: a single centered experience in Dhaka City. J. Natl. Inst. Neurosci. Bangladesh, 4: 54-57. https://doi.org/10.3329/jninb.v4i1.38287

Kumar A., Prasad J.K., Srivastava N., Ghosh S.K. 2019. Strategies to minimize various stress-related freezethaw damages during conventional cryopreservation of mammalian spermatozoa. Biopreserv. Biobank, 17: 603-612. https://doi.org/10.1089/bio.2019.0037

Lukefahr S. 2007. Strategies for the development of small- and extender-scale rabbit farming in South-East Asia. In Proc.: The International Conference on Rabbit Production, Bogor, Indonesia, 24-25.

Luong N.T., Thu D.V. 2005. Effects of Hagaton preparations on biological expenditures of coccidiosis in rabbits, J. Biol. Viet. Inst. Sci. Technol., 27: 78-81.

Magdanz V., Boryshpolets S., Ridzewski C., Eckel B., Reinhardt K. 2019. The motility-based swim-up technique separates bull sperm based on differences in metabolic rates and tail length. PloS ONE, 14: e0223576. https://doi.org/10.1371/journal.pone.0223576

Marri V., Richner H. 2014. Yolk carotenoids increase fledging success in great tit nestlings. Oecologia, 176: 371-377. https://doi.org/10.1007/s00442-014-3051-2

Nazif M.S., Rehman Z.U., Khan H., Khan F.A., Hussain T., Ahmad A., Farmanullah, Husnain A., Muhammad S., Murtaza G., Gang L. 2022. Glycine Improved Cryopreserved Spermatozoa Quality in Achai Bull. BioMed Res. Int., 2022: 8282387. https://doi.org/10.1155/2022/8282387

Ozimic S., Ban-Frangez H., Stimpfel M. 2023. Sperm Cryopreservation Today: Approaches, Efficiency, and Pitfalls. Curr. Issues Mol. Biol., 45: 4716-4734. https://doi.org/10.3390/cimb45060300

Palomar R.A., Gascón A., Martínez J.V., Balasch S., Molina Botella I. 2018. Sperm preparation after freezing improves motile sperm count, motility, and viability in frozen-thawed sperm compared with sperm preparation before freezingthawing process. J. Assist. Reprod. Genet., 35: 237-245. https://doi.org/10.1007/s10815-017-1050-z

Petyim S., Neungton C., Thanaboonyawat I., Laokirkkiat P., Choavaratana R. 2014. Sperm preparation before freezing improves sperm motility and reduces apoptosis in postfreezing-thawing sperm compared with post-thawing sperm preparation. J. Assist. Reprod. Genet., 31: 1673-1680. https://doi.org/10.1007/s10815-014-0332-y

Podico G., Ellerbrock R.E., Curcio B.R., Cheong S.H., Lima F.S., Canisso I.F. 2020. Single-Layer colloid centrifugation as a method to process urine-contaminated stallion semen after freezing-thawing. J. Equine Vet. Sci. 87: 102910. https://doi.org/10.1016/j.jevs.2020.102910

Rizkallah N., Chambers C.G., de Graaf S.P., Rickard J.P. 2022. Factors Affecting the Survival of Ram Spermatozoa during Liquid Storage and Options for Improvement. Animals., 12(3):244. https://doi.org/10.3390/ani12030244

Ros F., Offner S., Klostermann S., Thorey I., Niersbach H., Breuer S., Zarnt G., Lorenz S., Puels J., Siewe B., Schueler N., Dragicevic T., Ostler D., Hansen-Wester I., Lifke V., Kaluza B., Kaluza K., van Schooten W., Buelow R., Tissot A.C., Platzera J. 2020. Rabbits transgenic for human IgG genes recapitulating rabbit B-cell biology to generate human antibodies of high specificity and affinity. Mabs, 12: 1846900.

Shi L., Jin T., Hu Y., Ma Z., Niu H., Ren Y. 2020. Effects of reduced glutathione on ram sperm parameters, antioxidant status, mitochondrial activity and the abundance of hexose transporters during liquid storage at 5°C. Small Rumin. Res., 189: 106139. https://doi.org/10.1016/j.smallrumres.2020.106139

Silagadze Dali. 2022. Major Challenges of Agriculture. Georgian Scientists, 4: 145-153. Silvestre M.A., Yániz J.L., Peña F.J., Santolaria P., Castelló-Ruiz M. 2021. Role of Antioxidants in Cooled Liquid Storage of Mammal Spermatozoa. Antioxidants (Basel), 10: 1096. https://doi.org/10.3390/antiox10071096

Tam M.L., Thanh T.T.N., Tung T.N., Trung V.N., An T.T.N., Huy Q.V.N., Ngoc T.C. 2019. Cryopreservation of human spermatozoa by vitrification versus conventional rapid freezing: Effects on motility, viability, morphology and cellular defects. Eur. J. Obstet. Gyn. R. B., 234: 14-2. https://doi.org/10.1016/j.ejogrb.2019.01.001

Ugur M.R., Dinh T., Hitit M., Kaya, A., Topper E., Didion B., Memili E. 2020. Amino acids of seminal plasma associated with freezability of bull sperm. Front. Cell Dev. Biol, 7: 347. https://doi.org/10.3389/fcell.2019.00347

Wang A.W., Zhang H., Ikemoto I., Anderson D.J., Loughlin K.R. 1997. Reactive oxygen species generation by seminal cells during cryopreservation. Urology, 49: 921-925

Wang W., Wu Z., Dai Z., Yang Y., Wang J., Wu G. 2013. Glycine metabolism in animals and humans: implications for nutrition and health. Amino acids, 45: 463-477. https://doi.org/10.1007/s00726-013-1493-1

Zhang X.G., Liu Q., Wang L.Q., Yang G.S., Hu J.H. 2016. Effects of glutathione on sperm quality during liquid storage in boars. Anim. Sci. J., 87: 1195-1201. https://doi.org/10.1111/asj.12545

Zhong R.Z., Zhou D.W. 2013. Oxidative stress and role of natural plant derived antioxidants in animal reproduction. J. Integr. Agr., 12: 1826-1838. https://doi.org/10.1016/S2095-3119(13)60412-8

Downloads

Published

2023-12-26

Issue

Section

Reproduction