Technological advances in chicken semen cryopreservation: Impacts on sperm quality, freezability and fertility https://doi.org/10.12982/VIS.2026.089
Article Sidebar
Main Article Content
Abstract
To facilitate genetic diversity preservation and conservation initiatives, poultry semen cryopreservation remains as a critical technology in avian reproduction. Despite notable achievements in assisted reproductive technologies in the species in the past decades, the biochemical and physiological traits of chicken semen pose unique challenges for cryopreservation. This review examined the factors influencing the quality, fertility, and freezability of cryopreserved chicken semen while emphasizing on the role of proteins, antioxidants, and cryoprotectants like ethylene glycol and dimethylacetamide in maintaining sperm integrity. The fundamentals of cryopreservation, including important procedures like collection, dilution, cooling, and thawing were also discussed alongside advances in current techniques such as slow freezing and vitrification, and novel approaches like microfluidics and nanoparticle-based cryoprotection. The effects of lipid peroxidation and oxidative stress on sperm structure and DNA integrity during freezing was also elaborated, suggesting new cryoprotectants and antioxidants as mitigating measures. Breed-specific variations and environmental factors are also explored, underscoring the need for customized strategies to maximize cryopreservation results. The analysis also looked at promising technologies for improving cryopreservation outcomes such as artificial sperm reservoirs. This article also highlights the significance of ongoing research to improve the dependability and accessibility of semen cryopreservation for both commercial and conservation purposes, while addressing contemporary issues like standardization and ethical considerations. In conclusion, this comprehensive review of existing knowledge and emerging trends provides a roadmap for transformative research and development breakthroughs in poultry semen reproductive technologies.
Article Details
This work is licensed under a Creative Commons Attribution 4.0 International License.
Publishing an article with open access in Veterinary Integrative Sciences leaves the copyright with the author. The article is published under the Creative Commons Attribution License 4.0 (CC-BY 4.0), which allows users to read, copy, distribute and make derivative works from the material, as long as the author of the original work is cited.
References
Ahmed, H., Jahan, S., Riaz, M., Ijaz, M.U., Wahab, A., 2021. Improving the quality and in vitro fertilization rate of frozen-thawed semen of buffalo (Bubalus bubalis) bulls with the inclusion of vitamin B12 in the cryopreservation medium. Anim. Reprod. Sci. 229, 106761.
Akhtar, M.F., Ma, Q., Li, Y., Chai, W., Zhang, Z., Li, L.,Wang, C., 2022. Effect of sperm cryopreservation in farm animals using nanotechnology. Animals. 12, 2277.
Amesse, L.S., Srivastava, G., Uddin, D., Pfaff-Amesse, T., 2003. Comparison of cryopreserved sperm in vaporous and liquid nitrogen. J. Reprod. Med. 48, 319-324.
Appiah, M.O., He, B., Lu, W., Wang, J., 2019. Antioxidative effect of melatonin on cryopreserved chicken semen. Cryobiology. 89, 90-95.
Askarianzadeh, Z., Sharafi, M., Torshizi, M.A.K., 2018. Sperm quality characteristics and fertilization capacity after cryopreservation of rooster semen in extender exposed to a magnetic field. Anim. Reprod. Sci. 198, 37-46.
Bacon, L.D., Salter, D.W., Motta, J.V., Crittenden, L.B., Ogasawara, F.X., 1986. Cryopreservation of chicken semen of inbred or specialized strains. Poult. Sci. 65, 1965-1971.
Balogun, A.S., Narang, R., Cheema, R.S., Kumar, A., Singh, N., Vijaysingh, M., 2020. Comparison of conventional and automated freezing methods on PB2 rooster semen cryopreserved with glycerol and dimethylsulfoxide tris coconut-water extender. Bull. Natl. Res. Cent. 44, 1-8.
Barbas, J., Mascarenhas, R., 2009. Cryopreservation of domestic animal sperm cells. Cell Tissue Bank. 10, 49-62.
Behnamifar, A., Bernal, B., Torres, O., Luis-Chincoya, H., Ggil, M., García-Casado, P., Rahimi, S., Woelders, H., Santiago-Moreno, J., 2021. Research Note: Evaluation of two methods for adding cryoprotectant to semen and effects of bovine serum albumin on quality characteristics of cryopreserved rooster spermatozoa. Poult. Sci. 100, 101093.
Bernal, B., Castaño, C., Esteso, M., Toledano-Díaz, A., Domínguez-González, M., Gil, M., López-Sebastián, A., Campo, J., Blesbois, E., Santiago-Moreno, J., 2022. Birchen and Blue Leonesa sperm cryopreservation: a new technique for evaluating the integrity of cockerel sperm membranes. Br. Poult. Sci. 63, 244-251.
Bernal, B., Iglesias-Cabeza, N., Sánchez-Rivera, U., Toledano-Díaz, A., Castaño, C., Pérez-Cerezales, S., Gutiérrez-Adán, A., López-Sebastián, A., García-Casado, P., Gil, M. 2020. Effect of supplementation of valine to chicken extender on sperm cryoresistance and post-thaw fertilization capacity. Poult. Sci. 99, 7133-7141.
Blanco, J.M., Long, J.A., Gee, G., Wildt, D.E., Donoghue, A.M., 2012. Comparative cryopreservation of avian spermatozoa: Effects of freezing and thawing rates on turkey and sandhill crane sperm cryosurvival. Anim. Reprod. Sci. 131, 1-8.
Blank, M.H., Silva, V.C., Rui, B.R., Novaes, G.A., Castiglione, V.C., Pereira, R.J.G., 2020. Beneficial influence of fetal bovine serum on in vitro cryosurvival of chicken spermatozoa. Cryobiology. 95, 103-109.
Blesbois, E., Grasseau, I., Seigneurin, F., 2005. Membrane fluidity and the ability of domestic bird spermatozoa to survive cryopreservation. Reproduction. 129, 371-378.
Blesbois, E., Grasseau, I., Seigneurin, F., Mignon-Grasteau, S., Saint Jalme, M., Mialon-Richard, M., 2008. Predictors of success of semen cryopreservation in chickens. Theriogenology. 69, 252-261.
Blesbois, E., Seigneurin, F., Grasseau, I., Limouzin, C., Besnard, J., Gourichon, D., Coquerelle, G., Rault, P., Tixier-Boichard, M., 2007. Semen cryopreservation for ex situ management of genetic diversity in chicken: creation of the French avian cryobank. Poult. Sci. 86, 555-564.
Bongalhardo, D.C., 2025. Sperm fitness assessment in poultry: Brief review of in vitro methods. Anim. Reprod. Sci. 272, 107666.
Braun, J., Sakai, M., Hochi, S., Oguri, N., 1994. Preservation of ejaculated and epididymal stallion spermatozoa by cooling and freezing. Theriogenology. 41, 809-818.
Brouwers, J.F., Gadella, B.M., 2003. In situ detection and localization of lipid peroxidation in individual bovine sperm cells. Free Radic. Biol. Med. 35, 1382-1391.
Burrows, W., Quinn, J., 1937. The collection of spermatozoa from the domestic fowl and turkey. Poult. Sci. 16, 19-24.
Burt, C.T., Chalovich, J.M., 1978. Serine ethanolamine phosphodiester: a major component in chicken semen. Biochim. Biophys. Acta. 529, 186-188.
Chalah, T., Seigneurin, F., Blesbois, E., Brillard, J., 1999. In vitro comparison of fowl sperm viability in ejaculates frozen by three different techniques and relationship with subsequent fertility in vivo. Cryobiology. 39, 185-191.
Chatterjee, S., Gagnon, C., 2001. Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol. Reprod. Dev. 59, 451-458.
Chauychu-Noo, N., Thananurak, P., Boonkum, W., Vongpralub, T., Chankitisakul, V., 2021. Effect of organic selenium dietary supplementation on quality and fertility of cryopreserved chicken sperm. Cryobiology. 98, 57-62.
Cheng, C.Y., Chen, P.R., Chen, C.J., Wang, S.H., Chen, C.F., Lee, Y.P., Huang, S.Y., 2015. Differential protein expression in chicken spermatozoa before and after freezing–thawing treatment. Anim. Reprod. Sci. 152, 99-107.
Choi, J., Shin, D.B., Ko, Y.G., Do, Y.J., Seong, H.H., Kim, D.H., Kong, I.K., Kim, S., 2014. 59 Evaluation of cryopreserved chicken semen with different cryoprotectants. Reprod. Fertil. Dev. 26, 143-143.
Chuaychu-Noo, N., Thananurak, P., Chankitisakul, V., Vongpralub, T., 2017. Supplementing rooster sperm with Cholesterol-Loaded-Cyclodextrin improves fertility after cryopreservation. Cryobiology. 74, 8-12.
Çiftci, H., Aygün, A., 2018. Poultry semen cryopreservation technologies. World's Poult. Sci. J. 74, 699-710.
Cross, N.L., 2004. Reorganization of lipid rafts during capacitation of human sperm. Biol. Reprod. 71, 1367-1373.
Curry, M., Watson, P., 1994. Osmotic effects on ram and human sperm membranes in relation to thawing injury. Cryobiology. 31, 39-46.
Elomda, A.M., Mehaisen, G.M., Stino, F.K., Saad, M.F., Ghaly, M.M., Partyka, A., Abbas, A.O., Nassar, F.S., 2024. The characteristics of frozen-thawed rooster sperm using various intracellular cryoprotectants. Poult. Sci. 103, 104190.
Feyzi, S., Sharafi, M., Rahimi, S., 2018. Stress preconditioning of rooster semen before cryopreservation improves fertility potential of thawed sperm. Poult. Sci. 97, 2582-2590.
Fujihara, N., Howarth Jr, B., 1978. Lipid peroxidation in fowl spermatozoa. Poult. Sci. 57, 1766-1768.
Getachew, T., Goshu, G., Lemma, A., 2023. Effects of commercial and homemade extenders on post-thaw sperm quality and fertility of semen from Ethiopian Indigenous Horro Chicken Breed. World. Vet. J. 13, 341-347.
Giraud, M., Motta, C., Boucher, D., Grizard, G., 2000. Membrane fluidity predicts the outcome of cryopreservation of human spermatozoa. Hum. Reprod. 15, 2160-2164.
Gliozzi, T., Zaniboni, L., Cerolini, S., 2011. DNA fragmentation in chicken spermatozoa during cryopreservation. Theriogenology. 75, 1613-1622.
Grigg, G., Hodge, A., 1949. Electron microscopic studies of spermatozoa I. The morphology of the spermatozoon of the common domestic fowl (Gallus domesticus). Aust. J. Biol. Sci. 2, 271-286.
Guthrie, H., Welch, G., 2012. Effects of reactive oxygen species on sperm function. Theriogenology. 78, 1700-1708.
Hamad, S.K., Elomda, A.M., Sun, Y., Li, Y., Zong, Y., Chen, J., Abbas, A.O., Stino, F.K., Nazmi, A., Mehaisen, G.M., 2023. The in vitro evaluation of rooster semen pellets frozen with dimethylacetamide. Animals. 13, 1603.
Holt, W., 2000. Fundamental aspects of sperm cryobiology: the importance of species and individual differences. Theriogenology. 53, 47-58.
Hu, T., Taylor, L., Sherman, A., Tiambo, C.K., Kemp, S.J., Whitelaw, B., Hawken, R.J., Djikeng, A., Mcgrew, M.J., 2022. A low-tech, cost-effective and efficient method for safeguarding genetic diversity by direct cryopreservation of poultry embryonic reproductive cells. Elife. 11, e74036.
Karakus, F.N., Kuran, S.B., Solakoglu, S., 2021. Effect of curcumin on sperm parameters after the cryopreservation. Eur. J. Obstet. Gynecol. Reprod. Biol. 267, 161-166.
Khaeruddin, K., Junaedi, J., Hastuti, H., 2020. Cryopreservation of Indonesian native chicken semen by using dimethyl sulfoxide and various level of ethylene glycol as cryoprotectants. Biodiversitas. 21, 5718-5722.
Khaeruddin, K., Wahjuningsih, S., Ciptadi, G., Yusuf, M., Hermawansyah, H., Sahiruddin, S., 2022. Cryopreservation of Gaga’chicken semen from South Sulawesi, Indonesia with the addition of L-carnitine, hyaluronic acid, sucrose and their combination in diluent. Biodiversitas. 23, 3297-3302.
Kowalczyk, A., Kordan, W., 2024. Evaluation of the effectiveness of the use of exosomes in the regulation of the mitochondrial membrane potential of frozen/thawed spermatozoa. PLoS One. 19, e0303479.
Kucera, A.C., Heidinger, B.J., 2018. Avian semen collection by cloacal massage and isolation of DNA from sperm. J. Vis. Exp. 132, 55324.
Kumar, A., Singh, G., Jerome, A., Kumar, P., Arjun, V., Bala, R., Verma, N., Sharma, R., 2021. IGF-1 supplementation in semen affects mitochondrial functional and calcium status of buffalo sperm following cryopreservation. Anim. Reprod. Sci. 231, 106783.
Kumar, P., Pawaria, S., Dalal, J., Ravesh, S., Bharadwaj, S., Jerome, A., Kumar, D., Jan, M. H., Yadav, P., 2019. Sodium alginate potentiates antioxidants, cryoprotection and antibacterial activities of egg yolk extender during semen cryopreservation in buffalo. Anim. Reprod. Sci. 209, 106166.
Labas, V., Grasseau, I., Cahier, K., Gargaros, A., Harichaux, G., Teixeira-Gomes, A.P., Alves, S., Bourin, M., Gérard, N., Blesbois, E., 2015. Qualitative and quantitative peptidomic and proteomic approaches to phenotyping chicken semen. J. Proteom. 112, 313-335.
Ladha, S., 1998. Lipid heterogeneity and membrane fluidity in a highly polarized cell, the mammalian spermatozoon. J. Membr. Biol. 165, 1-10.
Lamirande, E.D., Gagnon, C., 1993. A positive role for the superoxide anion in triggering hyperactivation and capacitation of human spermatozoa. Int. J. Androl. 16, 21-25.
Larbi, A., Li, C., Quan, G., 2024. An updated review on the application of proteomics to explore sperm cryoinjury mechanisms in livestock animals. Anim. Reprod. Sci. 107441.
Lin, H.L. H., Blesbois, E., Vitorino Carvalho, A., 2022. Chicken semen cryopreservation: importance of cryoprotectants. World's Poult. Sci. J. 78, 139-160.
Long, J., 2006. Avian semen cryopreservation: what are the biological challenges? Poult. Sci. 85, 232-236.
Martin, G., Sabido, O., Durand, P., Levy, R., 2004. Cryopreservation induces an apoptosis-like mechanism in bull sperm. Biol. Reprod. 71, 28-37.
Masoudi, R., Esmaeilkhanian, S., Hatami, M., Sharafi, M., Hatefi, A., Zarei, F., Nateghi, R., Tohidkia, M.R., 2025. Supplementation of lake extender with cysteamine preserves quality parameters and fertility potential of post-thawed rooster sperm during cryopreservation process. BMC Vet. Res. 21, 21.
Medeiros, C., Forell, F., Oliveira, A., Rodrigues, J., 2002. Current status of sperm cryopreservation: why isn't it better? Theriogenology. 57, 327-344.
Mehaisen, G.M., Elomda, A.M., Hamad, S.K., Ghaly, M.M., Sun, Y., Li, Y., Zong, Y., Chen, J., Partyka, A., Nazmi, A., 2022. Effect of dimethylacetamide concentration on motility, quality, antioxidant biomarkers, anti-freeze gene expression, and fertilizing ability of frozen/thawed rooster sperm. Animals. 12, 2739.
Mehdipour, M., Daghigh Kia, H., Najafi, A., Mohammadi, H., Álvarez-Rodriguez, M., 2020a. Effect of crocin and naringenin supplementation in cryopreservation medium on post-thaw rooster sperm quality and expression of apoptosis associated genes. PLoS One. 15, e0241105.
Mehdipour, M., Kia, H.D., Martínez-Pastor, F., 2020b. Poloxamer 188 exerts a cryoprotective effect on rooster sperm and allows decreasing glycerol concentration in the freezing extender. Poult. Sci. 99, 6212-6220.
Mehdipour, M., Kia, H.D., Moghaddam, G., Hamishehkar, H., 2018. Effect of egg yolk plasma and soybean lecithin on rooster frozen-thawed sperm quality and fertility. Theriogenology. 116, 89-94.
Miranda, M., Kulíková, B., Vašíček, J., Olexiková, L., Iaffaldano, N., Chrenek, P., 2018. Effect of cryoprotectants and thawing temperatures on chicken sperm quality. Reprod. Domest. Anim. 53, 93-100.
Mocé, E., Grasseau, I., Blesbois, E., 2010. Cryoprotectant and freezing-process alter the ability of chicken sperm to acrosome react. Anim. Reprod. Sci. 122, 359-366.
Mohammad, M.S., Mardenli, O., Al-Tawash, A.S.A., 2021. Evaluation of the cryopreservation technology of poultry sperm: a review study. IOP Conf. Ser.Earth Environ. Sci. 735, 012016.
Moreira, S.S.J., Lago, A.E.d.A., Moura, A.A.A., Silva, A.R., 2022. Impact of seminal plasma composition on sperm freezability in wild mammals: a review. Biopreserv. Biobank. 20, 90-96.
Mosca, F., Madeddu, M., Sayed, A.A., Zaniboni, L., Iaffaldano, N., Cerolini, S., 2016. Combined effect of permeant and non-permeant cryoprotectants on the quality of frozen/thawed chicken sperm. Cryobiology. 73, 343-347.
Mosca, F., Zaniboni, L., Abdel Sayed, A., Iaffaldano, N., Soglia, D., Schiavone, A., Cerolini, S., 2020. Effect of N-Methylacetamide concentration and thawing rate on chicken sperm quality after cryopreservation. Animals. 10, 824.
Mosca, F., Zaniboni, L., Sayed, A.A., Madeddu, M., Iaffaldano, N., Cerolini, S., 2019. Effect of dimethylacetamide and N-methylacetamide on the quality and fertility of frozen/thawed chicken semen. Poult Sci. 98, 6071-6077.
Mphaphathi, M.L., Luseba, D., Sutherland, B., Nedambale, T.L., 2012. Comparison of slow freezing and vitrification methods for Venda cockerel’s spermatozoa. Open J. Anim. Sci. 2, 204-210.
Murugesan, S., Mahapatra, R., 2020. Cryopreservation of Ghagus chicken semen: Effect of cryoprotectants, diluents and thawing temperature. Reprod. Domest. Anim. 55, 951-957.
Mutalik, S., Salian, S.R., Avadhani, K., Menon, J., Joshi, H., Hegde, A.R., Kumar, P., Kalthur, G., Adiga, S.K., 2014. Liposome encapsulated soy lecithin and cholesterol can efficiently replace chicken egg yolk in human semen cryopreservation medium. Syst. Biol. Reprod. 60, 183-188.
Najafi, A., Kia, H.D., Mehdipour, M., Hamishehkar, H., Álvarez-Rodríguez, M., 2020. Effect of quercetin loaded liposomes or nanostructured lipid carrier (NLC) on post-thawed sperm quality and fertility of rooster sperm. Theriogenology. 152, 122-128.
Najafi, A., Mohammadi, H., Sharifi, S.D., Rahimi, A., 2024. Apigenin supplementation substantially improves rooster sperm freezability and post-thaw function. Sci. Rep. 14, 4527.
Najafi, A., Taheri, R.A., Mehdipour, M., Farnoosh, G., Martínez-Pastor, F., 2018. Lycopene-loaded nanoliposomes improve the performance of a modified Beltsville extender broiler breeder roosters. Anim. Reprod. Sci. 195, 168-175.
Nandi, S., Whyte, J., Taylor, L., Sherman, A., Nair, V., Kaiser, P., Mcgrew, M., 2016. Cryopreservation of specialized chicken lines using cultured primordial germ cells. Poult. Sci. 95, 1905-1911.
Nesci, S., Spinaci, M., Galeati, G., Nerozzi, C., Pagliarani, A., Algieri, C., Tamanini, C., Bucci, D., 2020. Sperm function and mitochondrial activity: an insight on boar sperm metabolism. Theriogenology. 144, 82-88.
Nguyen, T.M.D., Seigneurin, F., Froment, P., Combarnous, Y., Blesbois, E., 2015. The 5’-AMP-activated protein kinase (AMPK) is involved in the augmentation of antioxidant defenses in cryopreserved chicken sperm. PLoS One. 10, e0134420.
Nİzam, M.Y., Selçuk, M., 2021. Evaluation of rooster semen frozen with shilajit containing extender. In EDUVET International Veterinary Sciences Congress, Turkey, 25-27 June 2021, pp. 103.
Nonnis, S., Maffioli, E., Grana, J., Madeddu, M., Zaniboni, L., Marelli, S.P., Perricone, V., Iaffaldano, N., Tedeschi, G., Cerolini, S., 2025. Relation between proteome characterization and semen quality in Italian chicken breeds. PLoS One. 20, e0333802.
O’neill, H.C., Nikoloska, M., Ho, H., Doshi, A., Maalouf, W., 2019. Improved cryopreservation of spermatozoa using vitrification: comparison of cryoprotectants and a novel device for long-term storage. J. Assist. Reprod. Genet. 36, 1713-1720.
Oldenhof, H., Wolkers, W.F., Sieme, H., 2021. Cryopreservation of semen from domestic livestock: bovine, equine, and porcine sperm. In: Wolkers, W.F., Oldenhof, H. (Eds), Cryopreservation and freeze-drying protocols, 4th edition. Humana, New York.
Partyka, A., Łukaszewicz, E., Niżański, W., 2012. Effect of cryopreservation on sperm parameters, lipid peroxidation and antioxidant enzymes activity in fowl semen. Theriogenology. 77, 1497-1504.
Partyka, A., Niżański, W., Łukaszewicz, E., 2010. Evaluation of fresh and frozen-thawed fowl semen by flow cytometry. Theriogenology. 74, 1019-1027.
Partyka, A., Rodak, O., Bajzert, J., Kochan, J., Niżański, W., 2017. The effect of L‐carnitine, hypotaurine, and taurine supplementation on the quality of cryopreserved chicken semen. Biomed Res. Int. 2017, 7279341.
Partyka, A., Strojecki, M., Niżański, W., 2018. Cyclodextrins or cholesterol-loaded-cyclodextrins? A better choice for improved cryosurvival of chicken spermatozoa. Anim. Reprod. Sci. 93, 235-244.
Patel, K., Sharma, N., Mishra, V., Aggarwal, R., Suthar, A., Sheth, H., 2021. P–109 Comparison between the outcome of sperm vitrification protocol and conventional slow freezing protocol for semen cryopreservation. Hum. Reprod. 36, deab130.108.
Peláez, J., Bongalhardo, D., Long, J., 2011. Characterizing the glycocalyx of poultry spermatozoa: III. Semen cryopreservation methods alter the carbohydrate component of rooster sperm membrane glycoconjugates. Poult. Sci. 90, 435-443.
Peña, F. J., O’flaherty, C., Ortiz Rodríguez, J.M., Martín Cano, F.E., Gaitskell-Phillips, G.L., Gil, M.C., Ortega Ferrusola, C., 2019. Redox regulation and oxidative stress: The particular case of the stallion spermatozoa. Antioxidants. 8, 567.
Perez-Patiño, C., Barranco, I., Li, J., Padilla, L., Martinez, E.A., Rodriguez-Martinez, H., Roca, J., Parrilla, I., 2019. Cryopreservation differentially alters the proteome of epididymal and ejaculated pig spermatozoa. Int. J. Mol. Sci. 20, 1791.
Peris-Frau, P., Soler, A.J., Iniesta-Cuerda, M., Martín-Maestro, A., Sánchez-Ajofrín, I., Medina-Chávez, D.A., Fernández-Santos, M.R., García-Álvarez, O., Maroto-Morales, A., Montoro, V., 2020. Sperm cryodamage in ruminants: understanding the molecular changes induced by the cryopreservation process to optimize sperm quality. Int. J. Mol. Sci. 21, 2781.
Petričáková, K., Janošíková, M., Ptáček, M., Zita, L., Savvulidi, F.G., Partyka, A., 2022. Comparison of commercial poultry semen extenders modified for cryopreservation procedure in the genetic resource program of czech golden spotted hen. Animals. 12, 2886.
Polge, C., Smith, A.U., Parkes, A.S., 1949. Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature. 164, 666 (1949).
Rakha, B.A., Ansari, M., Akhter, S., Zafar, Z., Naseer, A., Hussain, I., Blesbois, E., Santiago-Moreno, J., 2018. Use of dimethylsulfoxide for semen cryopreservation in Indian red jungle fowl (Gallus gallus murghi). Theriogenology. 122, 61-67.
Rienzi, L., Gracia, C., Maggiulli, R., Labarbera, A.R., Kaser, D.J., Ubaldi, F.M., Vanderpoel, S., Racowsky, C., 2017. Oocyte, embryo and blastocyst cryopreservation in ART: systematic review and meta-analysis comparing slow-freezing versus vitrification to produce evidence for the development of global guidance. Hum. Reprod. Update. 23, 139-155.
Robbaani, M., Khaeruddin, K., Rachmawati, A., Nursita, I.W., Wahjuningsih, S., 2024. The use of ethylene glycol cryoprotectant with varying concentrations on the integrity of the plasma membrane and DNA damage in Gaga Chickens Semen. In The 11th International Conference of Innovation in Animal Science (ICIAS 2024), Malang, Indonesia, 4 November 2024, pp. 95-103.
Rui, B.R., Shibuya, F.Y., Kawaoku, A.J., Losano, J.D., Angrimani, D.S., Dalmazzo, A., Nichi, M., Pereira, R.J., 2017. Impact of induced levels of specific free radicals and malondialdehyde on chicken semen quality and fertility. Theriogenology. 90, 11-19.
Saadeldin, I.M., Khalil, W.A., Alharbi, M.G., Lee, S.H., 2020. The current trends in using nanoparticles, liposomes, and exosomes for semen cryopreservation. Animals. 10, 2281.
Saleh, F., Kocur, O., Xie, P., Rosenwaks, Z., Palermo, G., 2024. P-073 Microfluidic sperm selection yields competent spermatozoa with higher genomic integrity and cryostress tolerance. Hum. Reprod. 39, deae108. 449.
Salifu, A.R.S., Bautista, J.A., Rayos, A.A., Dizon, J.T., Sangel, P.P., 2023. Suitability of sugarcane extract as a local extender and the use of either DMSO or Glycerol as Cryoprotectant for the Cryopreservation of the Banaba Native Breed Chicken (Gallus gallus domesticus) Semen. Philipp. J. Sci. 152, 677-685.
Santiago-Moreno, J., Blesbois, E., 2020. Functional aspects of seminal plasma in bird reproduction. Int. J. Mol. Sci. 21, 5664.
Santiago-Moreno, J., Castaño, C., Toledano-Díaz, A., Coloma, M., López-Sebastián, A., Prieto, M., Campo, J., 2011. Semen cryopreservation for the creation of a Spanish poultry breeds cryobank: optimization of freezing rate and equilibration time. Poult. Sci. 90, 2047-2053.
Santiago-Moreno, J., Castaño, C., Toledano-Díaz, A., Esteso, M.C., López-Sebastián, A., Villaverde-Morcillo, S., Dávila, S., Gil, M., Blesbois, E., 2017. Successful chilling of red-legged partridge (Alectoris rufa) sperm for use in artificial insemination. Poult. Sci. 96, 4068-4074.
Sasaki, K., Tatsumi, T., Tsutsui, M., Niinomi, T., Imai, T., Naito, M., Tajima, A., Nishi, Y., 2010. A method for cryopreserving semen from Yakido roosters using N-methylacetamide as a cryoprotective agent. J. Poult. Sci. 47, 297-301.
Seigneurin, F., Blesbois, E., 1995. Effects of the freezing rate on viability and fertility of frozen-thawed fowl spermatozoa. Theriogenology. 43, 1351-1358.
Sexton, T., 1980. Optimal rates for cooling chicken semen from+ 5 to− 196 C. Poult. Sci. 59, 2765-2770.
Shahin, M.A., Khalil, W.A., Saadeldin, I.M., Swelum, A.A.A., El-Harairy, M.A., 2020. Comparison between the effects of adding vitamins, trace elements, and nanoparticles to shotor extender on the cryopreservation of dromedary camel epididymal spermatozoa. Animals. 10, 78.
Shanmugam, M., Mahapatra, R., 2019. Pellet method of semen cryopreservation: Effect of cryoprotectants, semen diluents and chicken lines. Braz. Arch. Biol. Technol. 62, e19180188.
Shanmugam, M., Mahapatra, R., 2022. Effect of tempol on post-thaw semen parameters and fertility during chicken semen cryopreservation. Available online: https://doi.org/10.1101/2022.03.31.486544.
Shi, H., Li, Q.Y., Li, H., Wang, H.Y., Fan, C.X., Dong, Q.Y., Pan, B.C., Ji, Z.L., Li, J.Y., 2024. ROS-induced oxidative stress is a major contributor to sperm cryoinjury. Hum. Reprod. 39, 310-325.
Shishova, N., Davydova, G., Kombarova, N., Davydova, I., Poltavtsev, A., Zaraisky, E., Poltavtseva, R., 2020. The use of nanosized liposomes from vegetable phospholipids in combination with albumine and some polysaccharides as cryoprotective agents in the course of cryopreservation. IOP Conf. Ser. Mater. Sci. Eng. 747, 012074.
Siegel, P., Beane, W., 1963. Semen characteristics of chickens maintained in all-male flocks and in individual cages. Poult. Sci. 42, 1028-1030.
Simoes, R., Feitosa, W.B., Siqueira, A.F.P., Nichi, M., Paula-Lopes, F.F., Marques, M.G., Peres, M.A., Barnabe, V.H., Visintin, J.A., Assumpção, M., 2013. Influence of bovine sperm DNA fragmentation and oxidative stress on early embryo in vitro development outcome. Reproduction. 146, 433-441.
Simsek, S., Hitit, M., Bodu, M., Memili, E., 2025. Sperm cell membranes of bulls and bucks associated with sperm fertility and freezability. Animals. 15, 3248.
Song, Y., Silversides, F., 2007. Production of offspring from cryopreserved chicken testicular tissue. Poult. Sci. 86, 1390-1396.
Sonseeda, P., Vongpralub, T., Laopaiboon, B., 2013. Effects of environmental factors, ages and breeds on semen characteristics in Thai indigenous chickens: A one-year study. Thai J. Vet. Med. 43, 347-352.
Stanishevskaya, O., Silyukova, Y., Pleshanov, N., Kurochkin, A., 2021. Role of mono-and disaccharide combination in cryoprotective medium for rooster semen to ensure cryoresistance of spermatozoa. Molecules. 26, 5920.
Sun, L., He, M., Wu, C., Zhang, S., Dai, J., Zhang, D., 2021. Beneficial influence of soybean lecithin nanoparticles on rooster frozen–thawed semen quality and fertility. Animals. 11, 1769.
Sun, Y., Xue, F., Li, Y., Fu, L., Bai, H., Ma, H., Xu, S., Chen, J., 2019. Differences in semen quality, testicular histomorphology, fertility, reproductive hormone levels, and expression of candidate genes according to sperm motility in Beijing-You chickens. Poult. Sci. 98, 4182-4189.
Surai, P., Blesbois, E., Grasseau, I., Chalah, T., Brillard, J.P., Wishart, G., Cerolini, S., Sparks, N., 1998. Fatty acid composition, glutathione peroxidase and superoxide dismutase activity and total antioxidant activity of avian semen. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 120, 527-533.
Suwimonteerabutr, J., Ketkaew, P., Netiprasert, G., Khaopong, C., Osiriphan, B., Sriamornrat, P., Nuntapaitoon, M., 2023. Supplementing semen extenders with a combination of phosphorus and vitamin B12 Improves post-thawed cryopreserved rooster semen quality. Front. Vet. Sci. 10, 1301186.
Suzuki, H., Lee, D.S., Sano, H., Akiyama, K., Suwa, Y., Abe, Y., 2007. Cryopreservation of canine spermatozoa in a simple extender composed of skim milk, glucose and glycerol. Biol. Reprod. 77, 237-238.
Svoradová, A., Kuželová, L., Vašíek, J., Baláži, A., Olexiková, L., Makarevich, A., Chrenek, P., 2021. Rooster spermatozoa cryopreservation and quality assessment. CryoLetters. 42, 59-66.
Taouzinet, L., Fatmi, S., Khellouf, A., Lahiani-Skiba, M., Skiba, M., Iguer-Ouada, M., 2022. Drug release, stability and efficiency of vitamin E loaded in liposomes for bovine sperm protection in cryopreservation medium. CryoLetters. 43, 50-57.
Thananurak, P., Chuaychu-Noo, N., Phasuk, Y., Vongpralub, T., 2020a. Comparison of TNC and standard extender on post-thaw quality and in vivo fertility of Thai native chicken sperm. Cryobiology. 92, 197-202.
Thananurak, P., Chuaychu-Noo, N., Thélie, A., Phasuk, Y., Vongpralub, T., Blesbois, E., 2019. Sucrose increases the quality and fertilizing ability of cryopreserved chicken sperms in contrast to raffinose. Poult. Sci. 98, 4161-4171.
Thananurak, P., Chuaychu-Noo, N., Thélie, A., Phasuk, Y., Vongpralub, T., Blesbois, E., 2020b. Different concentrations of cysteamine, ergothioneine, and serine modulate quality and fertilizing ability of cryopreserved chicken sperm. Poult. Sci. 99, 1185-1198.
Thélie, A., Bailliard, A., Seigneurin, F., Zerjal, T., Tixier-Boichard, M., Blesbois, E., 2019. Chicken semen cryopreservation and use for the restoration of rare genetic resources. Poult. Sci. 98, 447-455.
Tonus, C., Connan, D., Waroux, O., Vandenhove, B., Wayet, J., Gillet, L., Desmecht, D., Antoine, N., Ectors, F., Grobet, L. 2017. Cryopreservation of chicken primordial germ cells by vitrification and slow freezing: A comparative study. Theriogenology. 88, 197-206.
Torres, L., Hu, E., Tiersch, T.R., 2016. Cryopreservation in fish: current status and pathways to quality assurance and quality control in repository development. Reprod. Fertil. Dev. 28, 1105-1115.
Tselutin, K., Narubina, L., Mavrodina, T., Tur, B., 1995. Cryopreservation of poultry semen. Br. Poult. Sci. 36, 805-811.
Tselutin, K., Seigneurin, F., Blesbois, E., 1999. Comparison of cryoprotectants and methods of cryopreservation of fowl spermatozoa. Poult. Sci. 78, 586-590.
Ugur, M.R., Saber Abdelrahman, A., Evans, H.C., Gilmore, A.A., Hitit, M., Arifiantini, R.I., Purwantara, B., Kaya, A., Memili, E., 2019. Advances in cryopreservation of bull sperm. Front. Vet. Sci. 6, 268.
Ushiyama, A., Ishikawa, N., Tajima, A., Asano, A., 2016. Comparison of membrane characteristics between freshly ejaculated and cryopreserved sperm in the chicken. J. Poult. Sci. 53, 305-312.
Ushiyama, A., Tajima, A., Ishikawa, N., Asano, A., 2018. Modification of membrane cholesterol and desmosterol in chicken spermatozoa improves post-thaw survival and prevents impairment of sperm function after cryopreservation. Reprod. Fertil. Dev. 30, 591-599.
Wahjuningsih, S., Khaeruddin, Rachmawati, A., Tribudi, Y.A., Sari, A.P.Z.N.L., Wahju Nursita, I., Iustitia Sitaresmi, P., 2026. Pioneering comparative analysis of intracellular cryoprotectants on Gaga chicken sperm quality during cryopreservation. J. Appl. Anim. Res. 54, 2628620.
Wiebke, M., Hensel, B., Nitsche-Melkus, E., Jung, M., Schulze, M., 2022. Cooled storage of semen from livestock animals (part I): Boar, bull, and stallion. Anim. Reprod. Sci. 246, 106822.
Wishart, G., 1985. Quantitation of the fertilising ability of fresh compared with frozen and thawed fowl spermatozoa. Br. Poult. Sci. 26, 375-380.
Woelders, H., 2021. Cryopreservation of avian semen. In: Wolkers, W.F., Oldenhof, H. (Eds.), Cryopreservation and freeze-drying protocols, 4th edition. Humana, New York.
Wu, B., Yang, X., Yan, H., 2020. Improving the quality of rooster semen frozen in straws by screening the glycerol concentration and freezing rate. Br. Poult. Sci. 61, 173-179.
Xia, L., Lalli, M.F., Ansah, G.A., Buckland, R.B., 1988.
Ultrastructure of fresh and frozen-thawed spermatozoa of high and low fertility lines of chickens. Poult. Sci. 67, 819-825.
Yánez-Ortiz, I., Catalán, J., Rodríguez-Gil, J.E., Miró, J., Yeste, M., 2022. Advances in sperm cryopreservation in farm animals: Cattle, horse, pig and sheep. Anim. Reprod. Sci. 246, 106904.
Yata, V.K., Yadav, N., Katoch, V., Gangwar, D.K., Kumar, S., Mohanty, T.K., Prakash, B., Mohanty, A.K., 2022. Enrichment of motile spermatozoa from cattle semen samples by microfluidics method. Indian J. Anim. Sci. 92, 711-716.
Yousefi, M., Narchi, M., Sharafi, M., Borghei-Rad, S., Shahverdi, A., Masoudi, R., 2021. Rooster frozen-thawed semen quality following sublethal xanthine oxidase treatments. Anim. Reprod. Sci. 235, 106883.
Zaniboni, L., Cassinelli, C., Mangiagalli, M.G., Gliozzi, T.M., Cerolini, S., 2014. Pellet cryopreservation for chicken semen: Effects of sperm working concentration, cryoprotectant concentration, and equilibration time during in vitro processing. Theriogenology. 82, 251-258.
Zhu, H., Zhu, Y., Sun, C., Jiang, F., 2022. A preliminary study on the evaluation of human sperm head morphology with a domestic digital holographic image system. Phenomics. 2, 130-135.
Zong, Y., Li, Y., Sun, Y., Mehaisen, G.M., Ma, T., Chen, J., 2023. Chicken sperm cryopreservation: review of techniques, freezing damage, and freezability mechanisms. Agriculture. 13, 445.
Zong, Y., Sun, Y., Li, Y., Han, X., Ma, T., Zhao, Y., Yuan, J., Ma, H., Ma, L., Chen, J., 2024. Regulation of winter wheat-originated antifreeze glycoprotein on rooster spermatozoa freezability. Poult. Sci. 103, 104053.
Zong, Y., Sun, Y., Li, Y., Mehaisen, G.M., Yuan, J., Ma, H., Ni, A., Wang, Y., Hamad, S.K., Elomda, A.M., 2022. Effect of glycerol concentration, glycerol removal method, and straw type on the quality and fertility of frozen chicken semen. Poult. Sci. 101, 101840.
