2025-11-052025-11-052025CARNEIRO, Marcos Coura et al. Comparison of post-thaw motility and in vitro fertility between ejaculated and epididymal semen, and seminal cfDNA characterization in pantaneiro bulls. Biology, Basel, v. 14, n. 5, e465, 2025. DOI: 10.3390/biology14050465. Disponível em: https://www.mdpi.com/2079-7737/14/5/465. Acesso em: 3 nov. 2025.e- 2079-7737https://repositorio.bc.ufg.br//handle/ri/28957This study evaluated the post-thaw motility and in vitro fertility of ejaculated and epididymal semen from Pantaneiro bulls and characterized cell-free DNA (cfDNA) in fresh seminal plasma. Semen from five bulls was collected via electroejaculation or post-mortem epididymal extraction. Fresh semen parameters and cfDNA concentrations were assessed before cryopreservation. Post-thaw sperm kinetics were evaluated using CASA at 0 and 6 h of incubation, and in vitro embryo development was analyzed following IVF. Data were assessed using ANOVA and logistic regression. Ejaculate samples exhibited more morphological defects than epididymal samples (15.8% vs. 1.8%, p ≤ 0.05). Post-thaw, epididymal semen showed higher total (87.2% vs. 32.4%) and progressive (67.1% vs. 14.4%) motility at 0 h (p ≤ 0.05), and higher motility at 6 h (38.9% vs. 11.0%, p ≤ 0.05). In vitro fertility did not differ significantly between ejaculated (n = 525 oocytes) and epididymal (n = 500 oocytes) semen groups in terms of cleavage (49.6% vs. 44.2%) and blastocyst formation on D7 (26.1% vs. 22.2%, p > 0.05). cfDNA concentration in fresh semen ranged from 11.4 to 50.9 ng/µL. These findings indicate that epididymal sperm from Pantaneiro bulls retain high post-thaw motility and fertility. Additionally, cfDNA characterization in seminal plasma contributes to indigenous cattle preservation and advances in male fertility research.engAcesso Abertohttp://creativecommons.org/licenses/by-nc-nd/4.0/Idigenous cattleSemen cryopreservationIVPMolecular markerArtigo10.3390/biology14050465