{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T22:52:47Z","timestamp":1783032767363,"version":"3.54.6"},"reference-count":49,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2022,5,11]],"date-time":"2022-05-11T00:00:00Z","timestamp":1652227200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Forests Fund (Poland)","award":["OR.271.3.10.2017"],"award-info":[{"award-number":["OR.271.3.10.2017"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>The wisent, or European bison (Bison bonasus), belongs to the same family (Bovidae) as the American bison and domestic cattle. The wisent is the largest mammal in Europe, and is called the \u201cForest Emperor\u201d. The wisent is listed as \u201cVulnerable\u201d on the IUCN Red List, and is protected by international law. Achievements in reproductive biotechnology have opened new possibilities for the cryoconservation of the wisent germplasm. Therefore, this research aimed to improve a strategy for the protection and preservation of the European bison through the creation of a wisent germplasm bank, based on the following procedures: isolation and in vitro maturation (IVM) of oocytes, in vitro fertilization (IVF) of matured oocytes, in vitro embryo culture (IVC), and embryo cryopreservation. Wisent ovaries were isolated from females outside the reproductive season, and eliminated from breeding for reasons other than infertility. Cumulus\u2013oocyte complexes (COCs) were isolated from follicles greater than 2 mm in diameter and matured for 24 h and 30 h. After IVM, COCs were fertilized in vitro with wisent sperm. The obtained wisent zygotes, based on oocytes matured for 24 h and 30 h, were cultured for 216 h. Embryos at the morula and early blastocyst stages were vitrified and then warmed and transferred to interspecies recipients (Bos taurus). USG and biochemical tests were used to monitor pregnancies. This study obtained embryos in the morula and early blastocyst stages only after oocytes were fertilized and matured for 30 h. On average, per oocyte donor, 12.33 \u00b1 0.5 COCs were isolated, and only 9.33 \u00b1 0.61 COCs were qualified for in vitro maturation (75.68%), while 9.16 \u00b1 0.48 COCs were matured (84.32%). On average, per donor, 5.5 \u00b1 0.34 embryos were cleaved (59.96%) after 48 h post-fertilization (hpf), and 3.33 \u00b1 0.21 achieved the eight-cell stage (36.52%) after 96 hpf, while 1 \u00b1 0.21 morula and early blastocyst stages (10.71%) were achieved after 216 hpf. A total of six embryos (one morula and five early blastocysts) were obtained and vitrified; after warming, five of them were interspecies transferred to cattle (Bos taurus). On day 41 after fertilization, 3 out of 5 pregnancies were detected based on USG, P4, and PAG tests. However, no pregnancy was observed on day 86 after fertilization, indicating embryo resorption. This study shows that obtaining wisent embryos in vitro, and subsequent cryopreservation to create a wisent embryo bank, can be applied and implemented for the wisent protection program.<\/jats:p>","DOI":"10.3390\/ani12101239","type":"journal-article","created":{"date-parts":[[2022,5,11]],"date-time":"2022-05-11T10:19:46Z","timestamp":1652264386000},"page":"1239","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Establishment of a Wisent (Bison bonasus) Germplasm Bank"],"prefix":"10.3390","volume":"12","author":[{"given":"Anna Maria","family":"Duszewska","sequence":"first","affiliation":[{"name":"Department of Morphological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Nowoursynowska 159, 02-776 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Magdalena","family":"Baraniewicz-Ko\u0142ek","sequence":"additional","affiliation":[{"name":"Department of Morphological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Nowoursynowska 159, 02-776 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jaros\u0142aw","family":"Wojdan","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Nutrigenomics, Institute of Genetics and Animal Biotechnology, Polish Academy of Sciences, ul. Post\u0119pu 36A, Jastrz\u0119biec, 05-552 Magdalenka, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katarzyna","family":"Bar\u0142owska","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Nutrigenomics, Institute of Genetics and Animal Biotechnology, Polish Academy of Sciences, ul. Post\u0119pu 36A, Jastrz\u0119biec, 05-552 Magdalenka, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wojciech","family":"Bielecki","sequence":"additional","affiliation":[{"name":"Department of Pathology and Veterinary Diagnostics, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Nowoursynowska 159, 02-776 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pawe\u0142","family":"Gr\u0119da","sequence":"additional","affiliation":[{"name":"Department of Morphological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Nowoursynowska 159, 02-776 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1056-0784","authenticated-orcid":false,"given":"Wojciech","family":"Ni\u017ca\u0144ski","sequence":"additional","affiliation":[{"name":"Department of Reproduction and Clinic of Farm Animals, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life Sciences, 50-366 Wroclaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6166-3954","authenticated-orcid":false,"given":"Wanda","family":"Olech","sequence":"additional","affiliation":[{"name":"Department of Genetics and Animal Breeding, Faculty of Animal Science, Warsaw University of Life Sciences, 02-776 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,5,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1007\/s10344-018-1219-5","article-title":"Community attitudes to the European bison Bison bonasus in areas where its reintroduction is planned and in areas with existing populations in northeastern Poland","volume":"64","author":"Klich","year":"2018","journal-title":"Eur. 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