Preprint Article Version 1 This version is not peer-reviewed

Comparative Evaluation of Post-Thaw Viability and In Vitro Fertility Between Ejaculated and Epididymal Semen, and Characterization of Seminal cfDNA in Pantaneiro Bulls

Marcos Coura Carneiro
ORCID logo ,
Alice Caroline Souza Castro
,
Roberta Reis Silva
ORCID logo ,
José Eduardo Vieira Chaves
,
Venâncio Augusto Oliveira Silva
,
Natalia Ernandes Capobianco
,
Paulo José Bastos Queiroz
ORCID logo ,
Leonardo de França e Melo
ORCID logo ,
Eleonora Araújo Barbosa
,
Margot Alves Nunes Dode
,
José Felipe Warmling Spricigo
* ORCID logo
Version 1 : Received: 25 September 2024 / Approved: 25 September 2024 / Online: 25 September 2024 (12:08:13 CEST)

How to cite: Carneiro, M. C.; Castro, A. C. S.; Silva, R. R.; Chaves, J. E. V.; Silva, V. A. O.; Capobianco, N. E.; Queiroz, P. J. B.; Melo, L. D. F. E.; Barbosa, E. A.; Dode, M. A. N.; Spricigo, J. F. W. Comparative Evaluation of Post-Thaw Viability and In Vitro Fertility Between Ejaculated and Epididymal Semen, and Characterization of Seminal cfDNA in Pantaneiro Bulls. Preprints 2024, 2024091983. https://doi.org/10.20944/preprints202409.1983.v1 Carneiro, M. C.; Castro, A. C. S.; Silva, R. R.; Chaves, J. E. V.; Silva, V. A. O.; Capobianco, N. E.; Queiroz, P. J. B.; Melo, L. D. F. E.; Barbosa, E. A.; Dode, M. A. N.; Spricigo, J. F. W. Comparative Evaluation of Post-Thaw Viability and In Vitro Fertility Between Ejaculated and Epididymal Semen, and Characterization of Seminal cfDNA in Pantaneiro Bulls. Preprints 2024, 2024091983. https://doi.org/10.20944/preprints202409.1983.v1

Abstract

This study evaluated the post-thaw viability 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 hours 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 viability and fertility. Additionally, cfDNA characterization in seminal plasma contributes to indigenous cattle preservation and advances in male fertility research.

Keywords

Indigenous cattle; semen cryopreservation; IVP; molecular marker

Subject

Biology and Life Sciences, Biology and Biotechnology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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