{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T11:01:10Z","timestamp":1778756470918,"version":"3.51.4"},"reference-count":35,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2020,11,25]],"date-time":"2020-11-25T00:00:00Z","timestamp":1606262400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>The objectives of this study were to assess the cooling and freezing of donkey epididymal semen harvested immediately after castration (Experiment 1, n = 4) or after the shipment (24 or 48 h) of epididymides attached to testicles (Experiment 2, n = 14) or dissected apart (Experiment 3, n = 36). In each experiment, semen was frozen immediately (Non-Centrif) in an egg yolk-based semen extender (EY) or after processing through cushion-centrifugation (Centrif) while extended in a skim milk-based extender (SC). In all three experiments, cooled, pre-freeze, and post-thaw epididymal semen was assessed for total motility (TM), progressive motility (PM), plasma membrane integrity (PMI), and high mitochondrial membrane potential (HMMP). Data were analyzed with R using mixed models and Tukey\u2019s test as posthoc. Results showed that the cooling of epididymal semen up to 24 h after harvesting did not affect motility parameters or plasma membrane integrity; furthermore, in Experiment 3, the post-thaw evaluation of both Centrif and Non-Centrif achieved similar TM and PM. Collectively, the post-thaw results revealed low motility parameters across groups; while, the PMI and HMMP did not reflect this trend, and the values remained high, suggesting that there was a lack of epididymal sperm activation with either centrifugation or extenders. In summary, freshly harvested and cooled-shipped and cooled semen had satisfactory semen parameters. Future studies need to address donkey epididymal semen fertility in mares and jennies.<\/jats:p>","DOI":"10.3390\/ani10122209","type":"journal-article","created":{"date-parts":[[2020,11,25]],"date-time":"2020-11-25T08:59:15Z","timestamp":1606294755000},"page":"2209","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Donkey Epididymal Transport for Semen Cooling and Freezing"],"prefix":"10.3390","volume":"10","author":[{"given":"Yamilka","family":"Lago-Alvarez","sequence":"first","affiliation":[{"name":"Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8046-6254","authenticated-orcid":false,"given":"Giorgia","family":"Podico","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7526-7760","authenticated-orcid":false,"given":"Lorenzo G.","family":"Segabinazzi","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lais L.","family":"Cunha","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Leonardo","family":"Barbosa","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carolyn E.","family":"Arnold","sequence":"additional","affiliation":[{"name":"Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&amp;M University, College Station, TX 77840, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8377-6469","authenticated-orcid":false,"given":"Fabio S.","family":"Lima","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luise T.","family":"King","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Medicine, University of Arizona, Oro Valley, AZ 85704, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3523-2269","authenticated-orcid":false,"given":"Amy K.","family":"McLean","sequence":"additional","affiliation":[{"name":"Department of Animal Sciences, University of California, Davis, CA 95161, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Igor F.","family":"Canisso","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61801, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,11,25]]},"reference":[{"key":"ref_1","first-page":"0310","article-title":"Donkey breeding behavior with an emphasis on the P\u00eaga breed","volume":"20","author":"Canisso","year":"2010","journal-title":"Vet. Care Donkeys Int. Vet. Inf. Serv."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1016\/j.cveq.2019.08.014","article-title":"Key Aspects of Donkey and Mule Reproduction","volume":"35","author":"Canisso","year":"2019","journal-title":"Vet. Clin. N. Am. Equine Pract."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"652","DOI":"10.2746\/095777309X479058","article-title":"Strategies for the management of donkey jacks in intensive breeding systems","volume":"21","author":"Canisso","year":"2009","journal-title":"Equine Vet. Educ."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1111\/j.2042-3306.2010.00130.x","article-title":"Seminal parameters and field fertility of cryopreserved donkey jack semen after insemination of horse mares","volume":"43","author":"Canisso","year":"2011","journal-title":"Equine Vet. J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1846","DOI":"10.1016\/j.theriogenology.2012.07.015","article-title":"Donkey jack (Equus asinus) semen cryopreservation: Studies of seminal parameters, post-breeding inflammatory response, and fertility in donkey jennies","volume":"78","author":"Rota","year":"2012","journal-title":"Theriogenology"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.jevs.2016.11.005","article-title":"Comparison of Centrifugation and Noncentrifugation Methods to Cryopreserve Stallion Epididymal Semen","volume":"50","author":"Ellerbrock","year":"2017","journal-title":"J. Equine Vet. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"809","DOI":"10.1016\/0093-691X(94)90497-7","article-title":"Preservation of ejaculated and epididymal stallion spermatozoa","volume":"41","author":"Braun","year":"1994","journal-title":"Theriogenology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1016\/j.anireprosci.2008.05.003","article-title":"Freezing of stallion epididymal sperm","volume":"107","author":"Papa","year":"2008","journal-title":"Anim. Reprod. Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/S0737-0806(87)80025-4","article-title":"Principles of cryopreservation and a review of cryopreservation of stallion spermatozoa","volume":"7","author":"Amann","year":"1987","journal-title":"J. Equine Vet. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1016\/j.cveq.2006.08.007","article-title":"Collection and Freezing of Epididymal Stallion Sperm","volume":"22","author":"Bruemmer","year":"2006","journal-title":"Vet. Clin. N. Am. Equine Pract."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.jevs.2017.11.008","article-title":"The Current Situation and Trend of Donkey Industry in Europe","volume":"65","author":"Camillo","year":"2018","journal-title":"J. Equine Vet. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1016\/j.theriogenology.2008.09.005","article-title":"Artificial insemination in South American camelids and wild equids","volume":"71","author":"Adams","year":"2009","journal-title":"Theriogenology"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.repbio.2014.12.002","article-title":"A critical assessment of the effect of serine protease inhibitors on porcine fertilization and quality parameters of porcine spermatozoa in vitro","volume":"15","author":"Beek","year":"2015","journal-title":"Reprod. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1016\/S0093-691X(03)00256-5","article-title":"Epididymal sperm from the African buffalo (Syncerus caffer) can be frozen successfully with AndroMed\u00ae and with TriladylTM but the addition of bovine seminal plasma is detrimental","volume":"61","author":"Herold","year":"2004","journal-title":"Theriogenology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1089\/bio.2018.0021","article-title":"Effect of Extender and Freezing Rate on Quality Parameters and in Vitro Fertilization Capacity of Alpaca Spermatozoa Recovered from Cauda Epididymis","volume":"17","author":"Gonzales","year":"2019","journal-title":"Biopreserv. Biobank"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.theriogenology.2016.09.044","article-title":"Cryopreservation of bison epididymal sperm: A strategy for improving post-thaw quality when collecting sperm in field conditions","volume":"89","author":"Vilela","year":"2017","journal-title":"Theriogenology"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"102940","DOI":"10.1016\/j.jevs.2020.102940","article-title":"Preservation of Epididymal Stallion Sperm in Liquid and Frozen States: Effects of Seminal Plasma on Sperm Function and Fertility","volume":"88","author":"Vilagran","year":"2020","journal-title":"J. Equine Vet. Sci."},{"key":"ref_18","first-page":"485","article-title":"How to Process Stallion Epididymal Sperm in Your Practice","volume":"61","author":"Stawicki","year":"2015","journal-title":"Am. Assoc. Equine Pract."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"698","DOI":"10.1016\/j.theriogenology.2015.10.009","article-title":"Pregnancy outcomes using stallion epididymal sperm stored at 5 \u00b0C for 24 or 48 h before harvest","volume":"85","author":"Stawicki","year":"2016","journal-title":"Theriogenology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"704","DOI":"10.1016\/j.theriogenology.2008.04.047","article-title":"Factors impacting equine sperm recovery rate and quality following cushioned centrifugation","volume":"70","author":"Waite","year":"2008","journal-title":"Theriogenology"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1232","DOI":"10.1016\/j.theriogenology.2011.10.031","article-title":"The impact of cushioned centrifugation protocols on semen quality of stallions","volume":"77","author":"Bliss","year":"2012","journal-title":"Theriogenology"},{"key":"ref_22","unstructured":"Auer, J., and Stick, J. (2012). Equine Surgery, Elsevier."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1002\/cyto.b.21506","article-title":"A New Method for Evaluating Stallion Sperm Viability and Mitochondrial Membrane Potential in Fixed Semen Samples","volume":"94","author":"Ball","year":"2018","journal-title":"Cytom. Part B Clin. Cytom."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"845","DOI":"10.1111\/rda.12379","article-title":"Use of cholesterol-loaded cyclodextrin in donkey semen cryopreservation improves sperm viability but results in low fertility in mares","volume":"49","author":"Oliveira","year":"2014","journal-title":"Reprod. Domest. Anim."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Novello, G., Podico, G., Segabinazzi, L.G.T.M., Lima, F.S., and Canisso, I.F. (2020). Stallion Semen Cooling Using Native Phosphocaseinate-based Extender and Sodium Caseinate Cholesterol-loaded Cyclodextrin-based Extender. J. Equine Vet. Sci., 92.","DOI":"10.1016\/j.jevs.2020.103104"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"340","DOI":"10.1016\/j.theriogenology.2013.10.003","article-title":"Use of cholesterol-loaded cyclodextrin: An alternative for bad cooler stallions","volume":"81","author":"Hartwig","year":"2014","journal-title":"Theriogenology"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.cryobiol.2011.05.006","article-title":"Effects of cholesterol-loaded cyclodextrins on the quality of frozen-thawed equine epididymal sperm","volume":"63","author":"Pamornsakda","year":"2011","journal-title":"Cryobiology"},{"key":"ref_28","first-page":"302","article-title":"Glycerol in semen extender is a limiting factor in the fertility in asinine and equine species","volume":"89","author":"Vidament","year":"2005","journal-title":"Anim. Reprod. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.anireprosci.2008.03.016","article-title":"Differences in ability of jennies and mares to conceive with cooled and frozen semen containing glycerol or not","volume":"112","author":"Vidament","year":"2009","journal-title":"Anim. Reprod. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Saragusty, J., Lemma, A., Hildebrandt, T.B., and G\u00f6ritz, F. (2017). Follicular size predicts success in artificial insemination with frozen-Thawed sperm in donkeys. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0175637"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.jevs.2018.05.051","article-title":"Skim Milk, Casein Based Extender or Botucrio in Cooled Semen Asinus (Equus asinus)","volume":"66","author":"Rodrigues","year":"2018","journal-title":"J. Equine Vet. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1590\/S0102-09352013000300010","article-title":"Cooling of ejaculated and epididymal stallion sperm","volume":"65","author":"Monteiro","year":"2013","journal-title":"Arq. Bras. Med. Vet. Zootec."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1016\/S0093-691X(02)00797-5","article-title":"Capacitation dependent lipid rearrangements in the plasma membrane of equine sperm","volume":"58","author":"Colenbrander","year":"2002","journal-title":"Theriogenology"},{"key":"ref_34","first-page":"127","article-title":"Effect of cholesterol on the motility e plasma membrane integrity of frozen equine spermatozoa after thawing","volume":"56","author":"Combes","year":"2000","journal-title":"J. Reprod. Fertil. Suppl."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.jevs.2017.08.007","article-title":"Heterologous Oviductal Cells Binding Capacity of Cryopreserved Equine Ejaculated and Epididymal Spermatozoa","volume":"59","author":"Carneiro","year":"2017","journal-title":"J. Equine Vet. Sci."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/10\/12\/2209\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:37:15Z","timestamp":1760179035000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/10\/12\/2209"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,11,25]]},"references-count":35,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2020,12]]}},"alternative-id":["ani10122209"],"URL":"https:\/\/doi.org\/10.3390\/ani10122209","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,11,25]]}}}