{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T02:42:18Z","timestamp":1787020938677,"version":"build-2736575974"},"reference-count":59,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,10,12]],"date-time":"2024-10-12T00:00:00Z","timestamp":1728691200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,10,12]],"date-time":"2024-10-12T00:00:00Z","timestamp":1728691200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["J Animal Sci Biotechnol"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Sperm cryopreservation is widely used in the cattle industry, as it allows for disassociating the localization of sires and the collection of semen from the timing of artificial insemination. While freeze-thawing is known to impair sperm DNA integrity, whether the damage induced consists of single- (SSB) or double-strand breaks (DSB) has not been determined. In addition, no previous study has addressed if DNA breaks preferentially reside in specific genome regions such as those forming the toroid linker regions, or are rather spread throughout the regions linked to protamines. The main aim of the present work, therefore, was to elucidate the type and localization of the DNA damage generated by cryopreservation and to evaluate its impact on artificial insemination outcomes in cattle.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The incidence of SSB and DSB was evaluated in 12 ejaculates before and after cryopreservation with the Comet assay, and the localization of the DNA breaks was assessed using pulsed-field gel electrophoresis (PFGE). Before cryopreservation, the incidence of SSB was 10.99%\u2009\u00b1\u20094.62% and involved 20.56%\u2009\u00b1\u20093.04% of sperm cells, whereas these figures significantly (\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u20090.0001) increased up to 34.11%\u2009\u00b1\u20093.48% and 53.36%\u2009\u00b1\u200911.00% in frozen-thawed sperm. In contrast, no significant differences in the incidence of DSB were observed (\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&gt;\u20090.990) before and after cryopreservation (before: incidence of 13.91%\u2009\u00b1\u20091.75% of sperm DNA affecting 56.04%\u2009\u00b1\u200912.49% of sperm cells; after: incidence of 13.55%\u2009\u00b1\u20091.55% of sperm DNA involving 53.36%\u2009\u00b1\u200911.00% of sperm cells). Moreover, PFGE revealed that the percentage of sperm DNA fragments whose length was shorter than a toroid (&lt;\u200931.5\u00a0kb) was greater (\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u20090.0001) after (27.00%\u2009\u00b1\u20094.26%) than before freeze-thawing (15.57%\u2009\u00b1\u20094.53%). These differences indicated that the DNA breaks induced by cryopreservation affect the regions condensed in protamines, which are structured in toroids. On the other hand, in vivo fertility rates were associated to the incidence of SSB and DSB in frozen-thawed sperm (\n                      <jats:italic>P<\/jats:italic>\n                      \u2009=\u20090.032 and\n                      <jats:italic>P<\/jats:italic>\n                      \u2009=\u20090.005), but not with the size of the DNA fragments resulting from these breaks (\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&gt;\u20090.05).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Cryopreservation of bovine sperm generates single-strand DNA breaks, which are mainly located in protamine-condensed toroidal regions. The incidence of DNA breaks in cryopreserved sperm has an impact on cattle fertility, regardless of the size of generated fragments.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s40104-024-01099-0","type":"journal-article","created":{"date-parts":[[2024,10,11]],"date-time":"2024-10-11T21:01:31Z","timestamp":1728680491000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Cryopreservation of bovine sperm causes single-strand DNA breaks that are localized in the toroidal regions of chromatin"],"prefix":"10.1186","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9101-2044","authenticated-orcid":false,"given":"Jordi","family":"Ribas-Maynou","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodrigo","family":"Mui\u00f1o","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolina","family":"Tamargo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc","family":"Yeste","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,10,12]]},"reference":[{"key":"1099_CR1","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1016\/S0301-6226(03)00145-3","volume":"86","author":"JE Pryce","year":"2004","unstructured":"Pryce JE, Royal MD, Garnsworthy PC, Mao IL. Fertility in the high-producing dairy cow. Livest Prod Sci. 2004;86:125\u201335.","journal-title":"Livest Prod Sci"},{"key":"1099_CR2","doi-asserted-by":"publisher","first-page":"2285","DOI":"10.3168\/jds.S0022-0302(04)70049-1","volume":"87","author":"PM VanRaden","year":"2004","unstructured":"VanRaden PM, Sanders AH, Tooker ME, Miller RH, Norman HD, Kuhn MT, et al. Development of a national genetic evaluation for cow fertility. J Dairy Sci. 2004;87:2285\u201392.","journal-title":"J Dairy Sci"},{"key":"1099_CR3","doi-asserted-by":"publisher","first-page":"106904","DOI":"10.1016\/j.anireprosci.2021.106904","volume":"246","author":"I Y\u00e1nez-Ortiz","year":"2022","unstructured":"Y\u00e1nez-Ortiz I, Catal\u00e1n J, Rodr\u00edguez-Gil JE, Mir\u00f3 J, Yeste M. Advances in sperm cryopreservation in farm animals: Cattle, horse, pig and sheep. Anim Reprod Sci. 2022;246:106904.","journal-title":"Anim Reprod Sci"},{"key":"1099_CR4","doi-asserted-by":"publisher","first-page":"365","DOI":"10.1007\/978-1-0716-0783-1_15","volume":"2180","author":"H Oldenhof","year":"2021","unstructured":"Oldenhof H, Wolkers WF, Sieme H. Cryopreservation of semen from domestic livestock: Bovine, equine, and porcine sperm. Methods Mol Biol. 2021;2180:365\u201377.","journal-title":"Methods Mol Biol"},{"key":"1099_CR5","doi-asserted-by":"publisher","first-page":"S49","DOI":"10.1016\/j.ijvsm.2017.11.001","volume":"6","author":"WA Khalil","year":"2018","unstructured":"Khalil WA, El-Harairy MA, Zeidan AEB, Hassan MAE, Mohey-Elsaeed O. Evaluation of bull spermatozoa during and after cryopreservation: Structural and ultrastructural insights. Int J Vet Sci Med. 2018;6:S49\u201356.","journal-title":"Int J Vet Sci Med"},{"key":"1099_CR6","doi-asserted-by":"publisher","first-page":"655","DOI":"10.1111\/rda.13409","volume":"54","author":"LG Gr\u00f6tter","year":"2019","unstructured":"Gr\u00f6tter LG, Cattaneo L, Marini PE, Kjelland ME, Ferr\u00e9 LB. Recent advances in bovine sperm cryopreservation techniques with a focus on sperm post-thaw quality optimization. Reprod in Dom Anim. 2019;54:655\u201365.","journal-title":"Reprod in Dom Anim"},{"key":"1099_CR7","first-page":"1155","volume":"12","author":"S Bang","year":"2022","unstructured":"Bang S, Tanga BM, Fang X, Seong G, Saadeldin IM, Qamar AY, et al. Cryopreservation of pig semen using a quercetin-supplemented freezing extender. Life (Basel). 2022;12:1155.","journal-title":"Life (Basel)"},{"key":"1099_CR8","doi-asserted-by":"publisher","first-page":"4028","DOI":"10.3390\/cimb44090276","volume":"44","author":"HW Choi","year":"2022","unstructured":"Choi HW, Jang H. Application of nanoparticles and melatonin for cryopreservation of gametes and embryos. Curr Issues Mol Biol. 2022;44:4028\u201344.","journal-title":"Curr Issues Mol Biol"},{"key":"1099_CR9","doi-asserted-by":"crossref","unstructured":"Ngcobo JN, Ramukhithi FV, Nephawe KA, Mpofu TJ, Chokoe TC, Nedambale TL. Flaxseed oil as a source of omega n-3 fatty acids to improve semen quality from livestock animals: a review. Animals (Basel). 2021;11:3395.","DOI":"10.3390\/ani11123395"},{"key":"1099_CR10","doi-asserted-by":"crossref","unstructured":"Soltani L, Ghaneialvar H, Mahdavi AH. An overview of the role of metallic and nonmetallic nanoparticles and their salts during sperm cryopreservation and in vitro embryo manipulation. Nucleos Nucleot Nucl. 2023;42:262\u201379.","DOI":"10.1080\/15257770.2022.2124269"},{"key":"1099_CR11","first-page":"1","volume":"10","author":"IM Saadeldin","year":"2020","unstructured":"Saadeldin IM, Khalil WA, Alharbi MG, Lee SH. The current trends in using nanoparticles, liposomes, and exosomes for semen cryopreservation. Animals (Basel). 2020;10:2281.","journal-title":"Animals (Basel)"},{"key":"1099_CR12","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1262\/jrd.2021-079","volume":"68","author":"K Ogata","year":"2022","unstructured":"Ogata K, Imai A, Sato S, Nishino K, Watanabe S, Somfai T, et al. Effects of reduced glutathione supplementation in semen freezing extender on frozen-thawed bull semen and in vitro fertilization. J Reprod Dev. 2022;68:53\u201361.","journal-title":"J Reprod Dev"},{"key":"1099_CR13","doi-asserted-by":"publisher","first-page":"690","DOI":"10.1016\/j.theriogenology.2003.11.013","volume":"62","author":"J Gadea","year":"2004","unstructured":"Gadea J, Sell\u00e9s E, Marco MA, Coy P, Mat\u00e1s C, Romar R, et al. Decrease in glutathione content in boar sperm after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders. Theriogenology. 2004;62:690\u2013701.","journal-title":"Theriogenology"},{"key":"1099_CR14","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1126\/sciadv.aaz7602","volume":"6","author":"S Middelkamp","year":"2020","unstructured":"Middelkamp S, Van Tol HTA, Spierings DCJ, Boymans S, Guryev V, Roelen BAJ, et al. Sperm DNA damage causes genomic instability in early embryonic development. Sci Adv. 2020;6:eaaz7602.","journal-title":"Sci Adv"},{"key":"1099_CR15","doi-asserted-by":"publisher","first-page":"117","DOI":"10.1530\/REP-23-0011","volume":"166","author":"J Gao","year":"2023","unstructured":"Gao J, Yan Z, Yan L, Zhu X, Jiang H, Qiao J. The effect of sperm DNA fragmentation on the incidence and origin of whole and segmental chromosomal aneuploidies in human embryos. Reproduction. 2023;166:117\u201324.","journal-title":"Reproduction"},{"key":"1099_CR16","doi-asserted-by":"publisher","first-page":"103366","DOI":"10.1016\/j.rbmo.2023.103366","volume":"47","author":"W Fu","year":"2023","unstructured":"Fu W, Cui Q, Yang Z, Bu Z, Shi H, Bi B, et al. High sperm DNA fragmentation increased embryo aneuploidy rate in patients undergoing preimplantation genetic testing. Reprod Biomed Online. 2023;47:103366.","journal-title":"Reprod Biomed Online"},{"key":"1099_CR17","doi-asserted-by":"publisher","first-page":"635","DOI":"10.1093\/biolre\/ioad106","volume":"109","author":"H Nguyen","year":"2023","unstructured":"Nguyen H, Ribas-Maynou J, Wu H, Quon B, Inouye T, Walker B, et al. Low levels of mouse sperm chromatin fragmentation delay embryo development. Biol Reprod. 2023;109:635\u201343.","journal-title":"Biol Reprod"},{"key":"1099_CR18","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1186\/s40659-022-00386-2","volume":"55","author":"Y Mateo-Otero","year":"2022","unstructured":"Mateo-Otero Y, Llavanera M, Recuero S, Delgado-Berm\u00fadez A, Barranco I, Ribas-Maynou J, et al. Sperm DNA damage compromises embryo development, but not oocyte fertilisation in pigs. Biol Res. 2022;55:15.","journal-title":"Biol Res"},{"key":"1099_CR19","doi-asserted-by":"publisher","first-page":"745","DOI":"10.1007\/s10815-023-02748-5","volume":"40","author":"JG Alvarez","year":"2023","unstructured":"Alvarez JG, Garc\u00eda-Peir\u00f3 A, Barros A, Ferraz L, Sousa M, Sakkas D. Double strand DNA breaks in sperm: the bad guy in the crowd. J Assist Reprod Genet. 2023;40:745\u201351.","journal-title":"J Assist Reprod Genet"},{"key":"1099_CR20","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/ijms21113882","volume":"21","author":"A Agarwal","year":"2020","unstructured":"Agarwal A, Barb\u0103ro\u0219ie C, Ambar R, Finelli R. The impact of single-and double-strand DNA breaks in human spermatozoa on assisted reproduction. Int J Mol Sci. 2020;21:3882.","journal-title":"Int J Mol Sci"},{"key":"1099_CR21","doi-asserted-by":"publisher","first-page":"699","DOI":"10.1016\/j.fertnstert.2018.11.035","volume":"111","author":"A Casanovas","year":"2019","unstructured":"Casanovas A, Ribas-Maynou J, Lara-Cerrillo S, Jimenez-Macedo AR, Hortal O, Benet J, et al. Double-stranded sperm DNA damage is a cause of delay in embryo development and can impair implantation rates. Fertil Steril. 2019;111:699\u2013707.","journal-title":"Fertil Steril"},{"key":"1099_CR22","doi-asserted-by":"publisher","first-page":"105","DOI":"10.3390\/genes10020105","volume":"10","author":"J Ribas-Maynou","year":"2019","unstructured":"Ribas-Maynou J, Benet J. Single and double strand sperm DNA damage: different reproductive effects on male fertility. Genes (Basel). 2019;10:105.","journal-title":"Genes (Basel)"},{"key":"1099_CR23","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1007\/978-3-031-06573-6_10","volume":"70","author":"J Ribas-Maynou","year":"2022","unstructured":"Ribas-Maynou J, Nguyen H, Wu H, Ward WS. Functional aspects of sperm chromatin organization. Results Probl Cell Differ. 2022;70:295\u2013311.","journal-title":"Results Probl Cell Differ"},{"key":"1099_CR24","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1038\/aja.2010.75","volume":"13","author":"Y Yamauchi","year":"2011","unstructured":"Yamauchi Y, Shaman JA, Ward WS. Non-genetic contributions of the sperm nucleus to embryonic development. Asian J Androl. 2011;13:31\u20135.","journal-title":"Asian J Androl"},{"key":"1099_CR25","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1080\/01485010701329378","volume":"53","author":"JA Shaman","year":"2007","unstructured":"Shaman JA, Yamauchi Y, Ward WS. Function of the sperm nuclear matrix. Arch Androl. 2007;53:135\u201340.","journal-title":"Arch Androl"},{"key":"1099_CR26","doi-asserted-by":"publisher","first-page":"652","DOI":"10.1093\/humrep\/der461","volume":"27","author":"J Ribas-Maynou","year":"2012","unstructured":"Ribas-Maynou J, Garc\u00eda-Peir\u00f3 A, Abad C, Amengual MJ, Navarro J, Benet J. Alkaline and neutral comet assay profiles of sperm DNA damage in clinical groups. Hum Reprod. 2012;27:652\u20138.","journal-title":"Hum Reprod"},{"key":"1099_CR27","doi-asserted-by":"publisher","first-page":"gaac029","DOI":"10.1093\/molehr\/gaac029","volume":"29","author":"J Ribas-Maynou","year":"2022","unstructured":"Ribas-Maynou J, Nguyen H, Valle R, Wu H, Yeste M, Ward WS. Sperm degradation after vasectomy follows a sperm chromatin fragmentation-dependent mechanism causing DNA breaks in the toroid linker regions. Mol Hum Reprod. 2022;29:gaac029.","journal-title":"Mol Hum Reprod"},{"key":"1099_CR28","doi-asserted-by":"publisher","first-page":"105","DOI":"10.1186\/s40104-022-00754-8","volume":"13","author":"J Ribas-Maynou","year":"2022","unstructured":"Ribas-Maynou J, Delgado-Berm\u00fadez A, Mateo-Otero Y, Vi\u00f1olas E, Hidalgo CO, Ward WS, et al. Determination of double- and single-stranded DNA breaks in bovine sperm is predictive of their fertilizing capacity. J Anim Sci Biotechnol. 2022;13:105.","journal-title":"J Anim Sci Biotechnol"},{"key":"1099_CR29","doi-asserted-by":"publisher","first-page":"2437","DOI":"10.2527\/jas.2011-4492","volume":"90","author":"S Karoui","year":"2012","unstructured":"Karoui S, D\u00edaz C, Gonz\u00e1lez-Mar\u00edn C, Amenabar ME, Serrano M, Ugarte E, et al. Is sperm DNA fragmentation a good marker for field AI bull fertility? J Anim Sci. 2012;90:2437\u201349.","journal-title":"J Anim Sci"},{"key":"1099_CR30","doi-asserted-by":"publisher","first-page":"781","DOI":"10.1071\/RD06029","volume":"18","author":"KE Waterhouse","year":"2006","unstructured":"Waterhouse KE, Haugan T, Kommisrud E, Tverdal A, Flatberg G, Farstad W, et al. Sperm DNA damage is related to field fertility of semen from young norwegian red bulls. Reprod Fertil Dev. 2006;18:781\u20138.","journal-title":"Reprod Fertil Dev"},{"key":"1099_CR31","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1016\/j.anireprosci.2009.04.011","volume":"117","author":"KE Waterhouse","year":"2010","unstructured":"Waterhouse KE, Gjeldnes A, Tverdal A, De Angelis PM, Farstad W, H\u00e5\u00e5rd M, et al. Alterations of sperm DNA integrity during cryopreservation procedure and in vitro incubation of bull semen. Anim Reprod Sci. 2010;117:34\u201342.","journal-title":"Anim Reprod Sci"},{"key":"1099_CR32","doi-asserted-by":"publisher","first-page":"224","DOI":"10.1016\/j.theriogenology.2022.12.011","volume":"197","author":"J Gos\u00e1lvez","year":"2023","unstructured":"Gos\u00e1lvez J, Bartolom\u00e9-Nebreda J, Tvrd\u00e1 E, L\u00f3pez-Fern\u00e1ndez C, Fern\u00e1ndez JL, Johnston S. Bacterial DNase activity as a putative inductor of sperm DNA fragmentation in infected bull frozen-thawed semen samples. Theriogenology. 2023;197:224\u201331.","journal-title":"Theriogenology"},{"key":"1099_CR33","doi-asserted-by":"publisher","first-page":"562","DOI":"10.1016\/j.theriogenology.2016.02.007","volume":"86","author":"H G\u00fcrler","year":"2015","unstructured":"G\u00fcrler H, Malama E, Heppelmann M, Calisici O, Leiding C, Kastelic JP, et al. Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm. Theriogenology. 2015;86:562\u201371.","journal-title":"Theriogenology"},{"key":"1099_CR34","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1262\/jrd.2014-140","volume":"61","author":"K Takeda","year":"2015","unstructured":"Takeda K, Uchiyama K, Kinukawa M, Tagami T, Kaneda M, Watanabe S. Evaluation of sperm DNA damage in bulls by TUNEL assay as a parameter of semen quality. J Reprod and Dev. 2015;61:185\u201390.","journal-title":"J Reprod and Dev"},{"key":"1099_CR35","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1095\/biolreprod.103.024281","volume":"71","author":"G Martin","year":"2004","unstructured":"Martin G, Sabido O, Durand P, Levy R. Cryopreservation induces an apoptosis-like mechanism in bull sperm. Biol Reprod. 2004;71:28\u201337.","journal-title":"Biol Reprod"},{"key":"1099_CR36","doi-asserted-by":"publisher","first-page":"327","DOI":"10.1016\/j.rbmo.2018.05.012","volume":"37","author":"M Hezavehei","year":"2018","unstructured":"Hezavehei M, Sharafi M, Kouchesfahani HM, Henkel R, Agarwal A, Esmaeili V, et al. Sperm cryopreservation: A review on current molecular cryobiology and advanced approaches. Reprod Biomed Online. 2018;37:327\u201339.","journal-title":"Reprod Biomed Online"},{"key":"1099_CR37","doi-asserted-by":"publisher","first-page":"106329","DOI":"10.1016\/j.anireprosci.2020.106329","volume":"215","author":"I Ibanescu","year":"2020","unstructured":"Ibanescu I, Siuda M, Bollwein H. Motile sperm subpopulations in bull semen using different clustering approaches - Associations with flow cytometric sperm characteristics and fertility. Anim Reprod Sci. 2020;215:106329.","journal-title":"Anim Reprod Sci"},{"key":"1099_CR38","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1016\/j.anireprosci.2007.10.007","volume":"109","author":"R Mui\u00f1o","year":"2008","unstructured":"Mui\u00f1o R, Tamargo C, Hidalgo CO, Pe\u00f1a AI. Identification of sperm subpopulations with defined motility characteristics in ejaculates from Holstein bulls: effects of cryopreservation and between-bull variation. Anim Reprod Sci. 2008;109:27\u201339.","journal-title":"Anim Reprod Sci"},{"key":"1099_CR39","doi-asserted-by":"publisher","first-page":"462","DOI":"10.1095\/biolreprod65.2.462","volume":"65","author":"R Rathi","year":"2001","unstructured":"Rathi R, Colenbrander B, Bevers MM, Gadella BM. Evaluation of in vitro capacitation of stallion spermatozoa. Biol Reprod. 2001;65:462\u201370.","journal-title":"Biol Reprod"},{"key":"1099_CR40","doi-asserted-by":"publisher","first-page":"1089","DOI":"10.1152\/physrev.1999.79.4.1089","volume":"79","author":"A Takahashi","year":"1999","unstructured":"Takahashi A, Camacho P, Lechleiter JD, Herman B. Measurement of intracellular calcium. Physiol Rev. 1999;79:1089\u2013125.","journal-title":"Physiol Rev"},{"key":"1099_CR41","doi-asserted-by":"crossref","unstructured":"Garner DL, Thomas CA. Organelle-specific probe JC-1 identifies membrane potential differences in the mitochondrial function of bovine sperm. Mol Reprod Dev. 1999;53:222\u20139.","DOI":"10.1002\/(SICI)1098-2795(199906)53:2<222::AID-MRD11>3.0.CO;2-L"},{"key":"1099_CR42","doi-asserted-by":"publisher","first-page":"669182","DOI":"10.3389\/fcell.2021.669182","volume":"9","author":"J Ribas-Maynou","year":"2021","unstructured":"Ribas-Maynou J, Garcia-Bonavila E, Hidalgo CO, Catal\u00e1n J, Miro J, Yeste M. Species-specific differences in sperm chromatin decondensation between eutherian mammals underlie distinct lysis requirements. Front Cell Dev Biol. 2021;9:669182.","journal-title":"Front Cell Dev Biol"},{"key":"1099_CR43","doi-asserted-by":"publisher","first-page":"94","DOI":"10.1016\/j.theriogenology.2021.08.024","volume":"174","author":"J Ribas-Maynou","year":"2021","unstructured":"Ribas-Maynou J, Garcia-Bonavila E, Bonet S, Catal\u00e1n J, Salas-Huetos A, Yeste M. The TUNEL assay underestimates the incidence of DNA damage in pig sperm due to chromatin condensation. Theriogenology. 2021;174:94\u2013101.","journal-title":"Theriogenology"},{"key":"1099_CR44","doi-asserted-by":"publisher","first-page":"715","DOI":"10.1111\/j.2047-2927.2013.00111.x","volume":"1","author":"J Ribas-Maynou","year":"2013","unstructured":"Ribas-Maynou J, Garc\u00eda-Peir\u00f3 A, Fern\u00e1ndez-Encinas A, Abad C, Amengual MJJ, Prada E, et al. Comprehensive analysis of sperm DNA fragmentation by five different assays: TUNEL assay, SCSA, SCD test and alkaline and neutral Comet assay. Andrology. 2013;1:715\u201322.","journal-title":"Andrology"},{"key":"1099_CR45","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1530\/REP-10-0176","volume":"140","author":"P Villani","year":"2010","unstructured":"Villani P, Eleuteri P, Grollino MG, Rescia M, Altavista P, Span\u00f2 M, et al. Sperm DNA fragmentation induced by DNAse I and hydrogen peroxide: an in vitro comparative study among different mammalian species. Reproduction. 2010;140:445\u201352.","journal-title":"Reproduction"},{"key":"1099_CR46","doi-asserted-by":"publisher","first-page":"107094","DOI":"10.1016\/j.anireprosci.2022.107094","volume":"247","author":"L Bittner-Schwerda","year":"2022","unstructured":"Bittner-Schwerda L, Malama E, Siuda M, van Loon B, Bollwein H. The sperm chromatin structure assay does not detect alterations in sperm chromatin structure induced by hydrogen peroxide. Anim Reprod Sci. 2022;247:107094.","journal-title":"Anim Reprod Sci"},{"key":"1099_CR47","doi-asserted-by":"publisher","first-page":"100","DOI":"10.1016\/j.cryobiol.2007.10.176","volume":"56","author":"M S\u0142owi\u0144ska","year":"2008","unstructured":"S\u0142owi\u0144ska M, Karol H, Ciereszko A. Comet assay of fresh and cryopreserved bull spermatozoa. Cryobiology. 2008;56:100\u20132.","journal-title":"Cryobiology"},{"key":"1099_CR48","doi-asserted-by":"publisher","first-page":"83","DOI":"10.1111\/j.2047-2927.2013.00158.x","volume":"2","author":"J Ribas-Maynou","year":"2014","unstructured":"Ribas-Maynou J, Fern\u00e1ndez-Encinas A, Garc\u00eda-Peir\u00f3 A, Prada E, Abad C, Amengual MJ, et al. Human semen cryopreservation: a sperm DNA fragmentation study with alkaline and neutral Comet assay. Andrology. 2014;2:83\u20137.","journal-title":"Andrology"},{"key":"1099_CR49","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1016\/j.anireprosci.2016.06.010","volume":"172","author":"R Serafini","year":"2016","unstructured":"Serafini R, Love CC, Coletta A, Mari G, Mislei B, Caso C, et al. Sperm DNA integrity in frozen-thawed semen from Italian Mediterranean Buffalo bulls and its relationship to in vivo fertility. Anim Reprod Sci. 2016;172:26\u201331.","journal-title":"Anim Reprod Sci"},{"key":"1099_CR50","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1007\/s00441-005-1097-5","volume":"322","author":"S Lewis","year":"2005","unstructured":"Lewis S, Aitken R. DNA damage to spermatozoa has impacts on fertilization and pregnancy. Cell Tissue Res. 2005;322:33\u201341.","journal-title":"Cell Tissue Res"},{"key":"1099_CR51","doi-asserted-by":"publisher","first-page":"95","DOI":"10.3390\/antiox9020095","volume":"9","author":"JR Drevet","year":"2020","unstructured":"Drevet JR, Aitken RJ. Oxidation of sperm nucleus in mammals: a physiological necessity to some extent with adverse impacts on oocyte and offspring. Antioxidants. 2020;9:95.","journal-title":"Antioxidants"},{"key":"1099_CR52","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1007\/978-3-030-21664-1_7","volume":"1166","author":"JR Drevet","year":"2019","unstructured":"Drevet JR, Aitken RJ. Oxidative damage to sperm DNA: Attack and defense. Adv Exp Med Biol. 2019;1166:107\u201317.","journal-title":"Adv Exp Med Biol"},{"key":"1099_CR53","doi-asserted-by":"publisher","first-page":"451","DOI":"10.1002\/mrd.1052","volume":"59","author":"S Chatterjee","year":"2001","unstructured":"Chatterjee S, Gagnon C. Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol Reprod Dev. 2001;59:451\u20138.","journal-title":"Mol Reprod Dev"},{"key":"1099_CR54","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1016\/j.theriogenology.2017.01.011","volume":"92","author":"A Mostek","year":"2017","unstructured":"Mostek A, Dietrich MA, S\u0142owi\u0144ska M, Ciereszko A. Cryopreservation of bull semen is associated with carbonylation of sperm proteins. Theriogenology. 2017;92:95\u2013102.","journal-title":"Theriogenology"},{"key":"1099_CR55","doi-asserted-by":"publisher","first-page":"107215","DOI":"10.1016\/j.anireprosci.2023.107215","volume":"251","author":"V Sapanidou","year":"2023","unstructured":"Sapanidou V, Tsantarliotou MP, Lavrentiadou SN. A review of the use of antioxidants in bovine sperm preparation protocols. Anim Reprod Sci. 2023;251:107215.","journal-title":"Anim Reprod Sci"},{"key":"1099_CR56","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1186\/gb-2007-8-9-227","volume":"8","author":"R Balhorn","year":"2007","unstructured":"Balhorn R. The protamine family of sperm nuclear proteins. Genome Biol. 2007;8:227.","journal-title":"Genome Biol"},{"key":"1099_CR57","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1126\/science.286.5437.120","volume":"286","author":"LR Brewer","year":"1999","unstructured":"Brewer LR, Corzett M, Balhorn R. Protamine-induced condensation and decondensation of the same DNA molecule. Science. 1999;286:120\u20133.","journal-title":"Science"},{"key":"1099_CR58","doi-asserted-by":"publisher","first-page":"14925","DOI":"10.1073\/pnas.261560398","volume":"98","author":"NV Hud","year":"2001","unstructured":"Hud NV, Downing KH. Cryoelectron microscopy of lambda phage DNA condensates in vitreous ice: the fine structure of DNA toroids. Proc Natl Acad Sci U S A. 2001;98:14925\u201330.","journal-title":"Proc Natl Acad Sci U S A"},{"key":"1099_CR59","doi-asserted-by":"publisher","first-page":"272","DOI":"10.1002\/j.1939-4640.2005.tb01095.x","volume":"26","author":"B Sotolongo","year":"2005","unstructured":"Sotolongo B, Huang TTF, Isenberger E, Ward WS. An endogenous nuclease in hamster, mouse, and human spermatozoa cleaves DNA into loop-sized fragments. J Androl. 2005;26:272\u201380.","journal-title":"J Androl"}],"container-title":["Journal of Animal Science and Biotechnology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s40104-024-01099-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s40104-024-01099-0\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s40104-024-01099-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,10,11]],"date-time":"2024-10-11T21:02:40Z","timestamp":1728680560000},"score":1,"resource":{"primary":{"URL":"https:\/\/jasbsci.biomedcentral.com\/articles\/10.1186\/s40104-024-01099-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,12]]},"references-count":59,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["1099"],"URL":"https:\/\/doi.org\/10.1186\/s40104-024-01099-0","relation":{},"ISSN":["2049-1891"],"issn-type":[{"value":"2049-1891","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,12]]},"assertion":[{"value":"16 May 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"3 September 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"12 October 2024","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be defined as a potential conflict of interest.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"140"}}