{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T08:33:30Z","timestamp":1791362010417,"version":"4.3.1"},"reference-count":50,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2025,10,24]],"date-time":"2025-10-24T00:00:00Z","timestamp":1761264000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2025,10,24]],"date-time":"2025-10-24T00:00:00Z","timestamp":1761264000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microscopy Res &amp; Technique"],"published-print":{"date-parts":[[2026,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Sperm cryopreservation is a critical technique for improving the efficacy of artificial insemination in sheep breeding programs. This study evaluated the effects of different extenders and packaging methods on post\u2010thaw ram sperm quality, ultrastructure, and potential reproductive performance. Semen was collected from five healthy Ossimi rams and extended using three cryopreservation media: Tris\u2010egg yolk (Tris\u2010EY), Tris supplemented with 1% soybean lecithin (Tris\u2010SBL), or Tris supplemented with 2\u2009mM butylated hydroxytoluene (Tris\u2010BHT). The extended semen was then packaged in either plastic straws or pellets and cryopreserved in liquid nitrogen. Post\u2010thaw sperm quality was assessed by evaluating progressive motility, viability, and membrane integrity. Sperm ultrastructure was examined using transmission electron microscopy (TEM) and reproductive index. A two\u2010way ANOVA analysis was used to study the effects of the extender and the method of packing. Cryopreservation using straws significantly improved post\u2010thaw sperm quality compared to pellets, demonstrating higher progressive motility (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.003), viability (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.0001), and membrane integrity (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.0001). However, neither the packaging method nor the extender type significantly affected the plasma membrane status, acrosome integrity, or mitochondrial, tail, and nuclear damage (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&gt;\u20090.05). Ultrastructural analysis confirmed that straws, regardless of the extender used (Tris\u2010EY, Tris\u2010SBL, or Tris\u2010BHT), effectively preserved sperm ultrastructure, including the nucleus, head, acrosome, plasma membrane, and mitochondria. The methods of packaged and extender types did not significantly affect the conception rate and litter size in sheep (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&gt;\u20090.05). However, it is worth noting that Tris\u2010SBL in straws had the highest conception rate (85.7%) and litter size (1.5 per ewe), while Tris\u2010BHT in pellets had the lowest values (71.4% and 1.2, respectively). This study confirmed that plastic straw packaging significantly improved post\u2010thaw sperm quality. While Tris\u2010EY showed higher reproductive performance, further research is needed to establish statistical significance in pregnancy rates and litter size.\n                  <\/jats:p>","DOI":"10.1002\/jemt.70077","type":"journal-article","created":{"date-parts":[[2025,10,25]],"date-time":"2025-10-25T05:25:42Z","timestamp":1761369942000},"page":"481-495","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Effects of Extender and Packaging Devices on Ultrastructural Changes and Reproductive Index of Cryopreserved Spermatozoa of Ossimi Rams"],"prefix":"10.1002","volume":"89","author":[{"given":"Ahmed I.","family":"Yousif","sequence":"first","affiliation":[{"name":"Animal Production Research Institute Agriculture Research Centre, Ministry of Agriculture  Giza Dokki Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9539-3246","authenticated-orcid":false,"given":"Wael A.","family":"Khalil","sequence":"additional","affiliation":[{"name":"Department of Animal Production Faculty of Agriculture, Mansoura University  Mansoura Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abdel\u2010Khalek E.","family":"Abdel\u2010Khalek","sequence":"additional","affiliation":[{"name":"Department of Animal Production Faculty of Agriculture, Mansoura University  Mansoura Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bedir E.","family":"El\u2010Saidy","sequence":"additional","affiliation":[{"name":"Animal Production Research Institute Agriculture Research Centre, Ministry of Agriculture  Giza Dokki Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6873-0718","authenticated-orcid":false,"given":"Sameh A.","family":"Abdelnour","sequence":"additional","affiliation":[{"name":"Department of Animal Production Faculty of Agriculture, Zagazig University  Zagazig Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,10,24]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/rda.14139"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-023-37424-2"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.anireprosci.2024.107429"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.1080\/1828051X.2023.2231946"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.7831\/ras.11.0_171"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.anireprosci.2018.03.019"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.theriogenology.2004.04.010"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.5455\/ajvs.124422"},{"key":"e_1_2_12_10_1","unstructured":"Blomfield J. A.2010.Evaluation of Dietary Supplementation with Antioxidants on Fertility Parameters in Stallions The University of Waikato."},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10329-024-01136-6"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.theriogenology.2014.12.012"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1080\/10495398.2017.1334662"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.anireprosci.2024.107547"},{"key":"e_1_2_12_15_1","doi-asserted-by":"crossref","unstructured":"Dove H.2011.Sheep: Nutrition Management Encyclopedia of Animal Science Second Edition pp. 981\u2013983.","DOI":"10.1081\/E-EAS2-120045697"},{"key":"e_1_2_12_16_1","doi-asserted-by":"publisher","DOI":"10.21608\/ejrs.2015.46751"},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.1080\/1828051X.2024.2407081"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.21608\/ejvs.2024.248926.1671"},{"key":"e_1_2_12_19_1","doi-asserted-by":"publisher","DOI":"10.4103\/2305-0500.262834"},{"key":"e_1_2_12_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.repbio.2024.100912"},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.3390\/ani14101422"},{"key":"e_1_2_12_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.theriogenology.2023.05.032"},{"key":"e_1_2_12_23_1","doi-asserted-by":"publisher","DOI":"10.54203\/scil.2024.wvj40"},{"key":"e_1_2_12_24_1","doi-asserted-by":"publisher","DOI":"10.4103\/2305-0500.284277"},{"key":"e_1_2_12_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-025-10030-0"},{"key":"e_1_2_12_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.anireprosci.2024.107472"},{"key":"e_1_2_12_27_1","doi-asserted-by":"publisher","DOI":"10.1111\/rda.14485"},{"key":"e_1_2_12_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.theriogenology.2010.11.018"},{"key":"e_1_2_12_29_1","first-page":"7","article-title":"Effect of Different Concentration of Butylated Hydroxytoluene on the Quality of Boar Spermatozoa During Preservation at+ 5\u00b0C","volume":"9","author":"Linda N.","year":"2024","journal-title":"International Journal of Veterinary Sciences and Animal Husbandry"},{"key":"e_1_2_12_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cryobiol.2017.07.005"},{"key":"e_1_2_12_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.anireprosci.2023.107234"},{"key":"e_1_2_12_32_1","first-page":"343","article-title":"Electron Microscopy in the Study of Human Sperm Pathologies, Current Microscopy Contributions to Advances in Science and Technology","author":"Moretti E.","year":"2012","journal-title":"Antonio M\u00e9ndez\u2010Vilas"},{"key":"e_1_2_12_33_1","volume-title":"Alternatives to antibiotics in semen extenders used in artificial insemination, The Global Antimicrobial Resistance Epidemic\u2010Innovative Approaches and Cutting\u2010Edge Solutions","author":"Morrell J. M.","year":"2022"},{"key":"e_1_2_12_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cryobiol.2014.07.007"},{"key":"e_1_2_12_35_1","doi-asserted-by":"publisher","DOI":"10.3390\/agriculture12122001"},{"key":"e_1_2_12_36_1","doi-asserted-by":"publisher","DOI":"10.1002\/vms3.158"},{"key":"e_1_2_12_37_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1439-0531.2008.01284.x"},{"key":"e_1_2_12_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.xfnr.2020.09.002"},{"key":"e_1_2_12_39_1","doi-asserted-by":"publisher","DOI":"10.4142\/jvs.2011.12.3.267"},{"key":"e_1_2_12_40_1","doi-asserted-by":"publisher","DOI":"10.21451\/1984-3143-AR2018-0107"},{"key":"e_1_2_12_41_1","doi-asserted-by":"publisher","DOI":"10.3390\/antiox10071154"},{"key":"e_1_2_12_42_1","first-page":"149","article-title":"Ultrastructural Alterations in Sperm Heads Under Influence of Several Implementors to Ram Semen","volume":"42","author":"Pivko J.","year":"2009","journal-title":"Slovak Journal of Animal Science"},{"key":"e_1_2_12_43_1","doi-asserted-by":"publisher","DOI":"10.7717\/peerj.12447"},{"key":"e_1_2_12_44_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.fertnstert.2009.05.026"},{"key":"e_1_2_12_45_1","doi-asserted-by":"publisher","DOI":"10.1155\/2022\/7378379"},{"key":"e_1_2_12_46_1","first-page":"64","article-title":"Changes in Ultrastructure of Crossbred Bovine Spermatozoa in Butylated Hydroxytoluene Supplemented Semen","volume":"9","author":"Sharma M.","year":"2019","journal-title":"International Journal of Livestock Research"},{"key":"e_1_2_12_47_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodhyd.2019.105510"},{"key":"e_1_2_12_48_1","doi-asserted-by":"publisher","DOI":"10.4103\/apjr.apjr_93_23"},{"key":"e_1_2_12_49_1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00009.2020"},{"key":"e_1_2_12_50_1","first-page":"257","article-title":"Biology of Sperm Capacitation: Evidence for Multiple Signalling Pathways","volume":"63","author":"Tulsiani D. R.","year":"2007","journal-title":"Society of Reproduction and Fertility Supplement"},{"key":"e_1_2_12_51_1","doi-asserted-by":"publisher","DOI":"10.5713\/ajas.20.0118"}],"container-title":["Microscopy Research and Technique"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jemt.70077","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/jemt.70077","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jemt.70077","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T16:07:24Z","timestamp":1770998844000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/jemt.70077"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,24]]},"references-count":50,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2026,3]]}},"alternative-id":["10.1002\/jemt.70077"],"URL":"https:\/\/doi.org\/10.1002\/jemt.70077","archive":["Portico"],"relation":{},"ISSN":["1059-910X","1097-0029"],"issn-type":[{"value":"1059-910X","type":"print"},{"value":"1097-0029","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,24]]},"assertion":[{"value":"2025-03-17","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-09-13","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-10-24","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}