Study of sericin added to semen extender in sperm cryopreservation of Hungarian Giant Rabbit
Keywords:
rabbit sperm, cryopreservation, sericin, Hungarian GiantAbstract
Because of the dramatic decline of Hungarian Giant rabbit we aim to create an in vitro sperm bank for safe gene preservation purposes. During testing different freezing protocols we developed an effective extender for the cryopreservation of Hungarian Giant sperm. The present study investigated the effect of sericin as antioxidants on sperm freezability using the modified Lake’s extender. Collected sperm from 10 bucks was diluted 1:4 with modified Lake’s extender with 1% glycerol and 4% DMF (control group), containing sericin 0.1 and 0.3 w/v% (treated groups). Samples filled into 0.25 ml straws, and after equilibration at 5°C for 60 min static freezing (5 cm above liquid nitrogen for 15 minutes) was applied. The thawing was carried out at 40ºC for 30 sec. In the fresh and frozen/thawed sperm samples motility (CASA) and ratio of live/dead cells, sperm abnormalities (aniline blue-eosin staining) were checked. In testing two different sericin concentrations no significant difference was observed in the proportion of live, intact cells or in total and progressive motility, none of the tested concentrations of sericin supplementation improved sperm viability. Further studies are required to determine whether the concentrations were suboptimal or this rabbit breed responds poorly to these treatments.
References
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BÁNYÁSZ, Á. – DROBNYÁK, Á. – KISSNÉ VÁRADI, É. – SZABÓ, ZS. – TÖRÖK, É. – BARNA, J. – VÉGI, B. (2025): Effects of L-Carnitine and Sericin on semen cryopreservation in roosters. 61st meeting Society for Low Temperature Biology, Olsztyn, Poland, 11-13 September 2025. p.10.
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KISSNÉ VÁRADI, É. - DROBNYÁK, Á. - BARNA, J. - TÖRÖK, É. SZABÓ, ZS. - VÉGI, B. (2026): Preliminary studies for elaborating a semen cryopreservation protocol of the Hungarian Giant rabbit. Anim. Reprod. Sci. 291, 108233.
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PAUL, D.R. - TALUKDAR, D. - AHMED, F.A. - LALRINTLUANGA, K. - KALITA, G. - TOLENKHOMBA, T.C. - CHAKRAVARTY, H. - KATIYAR, R. - KHARGHARIA, G. - DEORI, S. (2024): Effect of selenium nanoparticles on the quality and fertility of short-term preserved boar semen. Front. Vet. Sci. 10, 1-7.
PEGG, D.E. (2014): Principles of Cryopreservation. Methods Mol. Biol. 3–19.
RATCHAMAK, R. - RATSIRI, T. - KHEAWKANHA, T. - VONGPRALUB, T. - BOONKUM, W. – CHANKITISAKUL, V. (2020): Evaluation of cryopreserved boar semen after supplementation sericin from silkworm (Bombyx mori) in semen extender. Anim. Sci. J. 91(1), 1-7.
RAZA, S. - UCAN, U. - AKSOY, M. - ERDOGAN, G. - CEYLAN, A. - SERIN, I. (2019): Silk protein sericin pretreatment enhances osmotic tolerance and post-thaw sperm quality but reduces the ability of sperm cells to undergo in vitro induced acrosome reaction in rabbit. Cryobiology, 90, 1–7.
ROSATO, M.P. – IAFFALDANO, N. (2011): Effect of chilling temperature on the longterm survival of rabbit spermatozoa held either in a Tris-based or a jellified extender. Reprod. Dom. Anim. 46, 301–308.
VÁRADI, É. - VÉGI, B. - LIPTÓI, K. - BARNA, J. (2013): Methods for Cryopreservation of Guinea Fowl Sperm. Plos One, April 2013 Vol. 8 Issue 4 e62759.
VIJYETA, H.P. - KAVANI, F.S. - VAGH, A.A. - SAOLANKI, G.B. - VALA, K.B. - SINGH, V.K. - DHAM, A.J. (2024): Effect of supplementation of antioxidant sericin in semen extender on post-thaw sperm quality, oxidative status, and fertility of Jaffarabadi buffalo bull semen. Indian J. Vet. Sci. Biotechnol. 20(2), 79-84.
YANGNGAM, Y. - CHAPANYA, S. - VONGPRALUB, T. - BOONKUM, W. - CHANKITISAKUL, V. (2021): Effect of semen extender supplementation with sericin on post-thaw dairy bull sperm quality and lipid peroxidation. Czech. J. Anim. Sci. 66(1), 13–20.
ZHANG, Y.Q. (2002): Applications of natural silk protein sericin in biomaterials. Biotechnol. Adv. 20, 91–100.
AKSOY, M. - CANKAT LEHIMCIOGLU, N. - AKMAN, O. (2010): Effect of seminal plasma on functional integrity of rabbit sperm membranes during storage at 4°C or freezing. World Rabbit Sci. 16.
AKSOY, D. - TURAN, D.N. - BAYRAKTAR, Z.B. (2024): Cholesterol and Sericin as First Aid for Damaged Cells. J. Biosci. Med. 12, 79–88.
ALI, MD.Y. – KHATUN, A. (2026): Sericin in semen cryopreservation: A review of its potential and challenes. Vet. Integr Sci. 24(1), e2026019-1-23.
BÁNYÁSZ, Á. – DROBNYÁK, Á. – KISSNÉ VÁRADI, É. – SZABÓ, ZS. – TÖRÖK, É. – BARNA, J. – VÉGI, B. (2025): Effects of L-Carnitine and Sericin on semen cryopreservation in roosters. 61st meeting Society for Low Temperature Biology, Olsztyn, Poland, 11-13 September 2025. p.10.
CHATURVEDI, D. - DHAMI, A.J. - CHAUDHARI, D.V. (2022): Fortification of tris extender with mifepristone, sericin and taurine improves velocity and kinematics of fresh and frozen-thawed bovine spermatozoa. Indian J. Anim. Res. 56(3), 255–262.
DI IORIO, M. - LAURIOLA, F. - RUSCO, G. - ANTENUCCI, E. - SCHIAVITTO, M. - IAFFALDANO, N. (2024): Cryopreserving Rabbit Semen: Impact of Varying Sperm Concentrations on Quality and the Standardization of Protocol. Vet. Sci. 11(1), 9.
GHASEMI, M. – FARSHAD, A. HAJARIAN, H. – BANAFSHI, O. – ASADOLLAHI, V. – FATHI, F. (2018): The effects of sericin on cryopreserved sperm cells and subsequent embryo development in mice. Int. J. Reprod. Biomed. 16(6), 405-412.
KAMAL, M.M. - ALAM, M.E. - DAS, S.K. - YEASMIN, M.S. - AHMED, S. - RAHMAN, M.A. - DAS, D.K. - GOFUR, M.R. (2023): Effects of glucose and trehalose on tris-citric acid-egg yolk-fructose diluents for semen cryopreservation in goat. J. Adv. Vet. Anim. Res. 10, 169–177.
KARIMI‐SABET, M.J. - KHODAEI‐MOTLAGH, M. - MASOUDI, R. - SHARAFI, M. (2024): Zinc oxide nanoparticles preserve the quality and fertility potential of rooster sperm during the cryopreservation process. Reprod. Domest. Anim. 59(4), 1-7.
KATO, Y. - SASAKI, M. - YAMADA, H. - TERADA, S. (2005): Development of a novel serum-free freezing medium for mammalian cells using the silk protein sericin. Biotechnol. Appl. Biochem. 42(2), 183.
KHAN, R.U. (2011): Antioxidants and poultry semen quality. World’s Poultry Science J. 67 (2), 297-308.
KISSNÉ VÁRADI, É. - DROBNYÁK, Á. - BARNA, J. - TÖRÖK, É. SZABÓ, ZS. - VÉGI, B. (2026): Preliminary studies for elaborating a semen cryopreservation protocol of the Hungarian Giant rabbit. Anim. Reprod. Sci. 291, 108233.
KUMAR, P. - KUMAR, D. - SIKKA, P. - SINGH, P. (2015): Sericin supplementation improves semen freezabilIty of buffalo bulls by minimizing oxidative stress during cryopreservation. Anim. Reprod. Sci. 152, 26–31.
KUMAR, S. - GUPTA, H.P. - PRASAD, S. - AMBWANI, T.K. - SHARMA, R.K. - SINGH, J.L. (2025): Studies on the quality and antioxidant status of cryopreserved Pantja buck semen supplemented with aqueous extract of rosemary and sericin in tris extender. Biopreserv. Biobank. 23(4), 354-363.
KUNDU, S.C. - DASH, B.C. - DASH, R. - KAPLAN, D.L. (2008): Natural protective glue protein, sericin bioengineered by silkworms: Potential for biomedical and biotechnological applications. Prog. Polym. Sci. 33(10), 998–1012.
NASIRABADI, H.M. – SHIRAZI, A. – KADIVAR, A. - SHAMS-ESFANDABADI, N. – MOHEBBI, A. – AHMADI, E. (2019): Sericin Ameliorates the Capacitation State and Chromatin Integrity of Frozen-Thawed Stallion Spermatozoa by Reducing Oxidative Stress. Avicenna. J. Med. Biotechnol. 11(3), 245-252.
PAUL, D.R. - TALUKDAR, D. - AHMED, F.A. - LALRINTLUANGA, K. - KALITA, G. - TOLENKHOMBA, T.C. - CHAKRAVARTY, H. - KATIYAR, R. - KHARGHARIA, G. - DEORI, S. (2024): Effect of selenium nanoparticles on the quality and fertility of short-term preserved boar semen. Front. Vet. Sci. 10, 1-7.
PEGG, D.E. (2014): Principles of Cryopreservation. Methods Mol. Biol. 3–19.
RATCHAMAK, R. - RATSIRI, T. - KHEAWKANHA, T. - VONGPRALUB, T. - BOONKUM, W. – CHANKITISAKUL, V. (2020): Evaluation of cryopreserved boar semen after supplementation sericin from silkworm (Bombyx mori) in semen extender. Anim. Sci. J. 91(1), 1-7.
RAZA, S. - UCAN, U. - AKSOY, M. - ERDOGAN, G. - CEYLAN, A. - SERIN, I. (2019): Silk protein sericin pretreatment enhances osmotic tolerance and post-thaw sperm quality but reduces the ability of sperm cells to undergo in vitro induced acrosome reaction in rabbit. Cryobiology, 90, 1–7.
ROSATO, M.P. – IAFFALDANO, N. (2011): Effect of chilling temperature on the longterm survival of rabbit spermatozoa held either in a Tris-based or a jellified extender. Reprod. Dom. Anim. 46, 301–308.
VÁRADI, É. - VÉGI, B. - LIPTÓI, K. - BARNA, J. (2013): Methods for Cryopreservation of Guinea Fowl Sperm. Plos One, April 2013 Vol. 8 Issue 4 e62759.
VIJYETA, H.P. - KAVANI, F.S. - VAGH, A.A. - SAOLANKI, G.B. - VALA, K.B. - SINGH, V.K. - DHAM, A.J. (2024): Effect of supplementation of antioxidant sericin in semen extender on post-thaw sperm quality, oxidative status, and fertility of Jaffarabadi buffalo bull semen. Indian J. Vet. Sci. Biotechnol. 20(2), 79-84.
YANGNGAM, Y. - CHAPANYA, S. - VONGPRALUB, T. - BOONKUM, W. - CHANKITISAKUL, V. (2021): Effect of semen extender supplementation with sericin on post-thaw dairy bull sperm quality and lipid peroxidation. Czech. J. Anim. Sci. 66(1), 13–20.
ZHANG, Y.Q. (2002): Applications of natural silk protein sericin in biomaterials. Biotechnol. Adv. 20, 91–100.
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2026-11-09
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