Variable Inter-assay Estimation of Sperm DNA Fragmentation In Stallions Classified as Good and Bad Semen Freezers.
Abstract: Semen cryopreservation causes DNA damage, thus requiring continuous monitoring. Objective: To compare two assays for sperm DNA fragmentation (SDF) from stallions with contrasting semen freezability. Methods: Thirteen stallions were classified as good semen freezers (GSF) or bad semen freezers (BSF). Ejaculates were cryopreserved with three diluents. Semen was subject to SDF evaluation using the sperm chromatin structure assay (SCSA) and Halomax after thawing (0 h) and after a 4 h thermoresistance test. Results: On semen of BSF, analysis by SCSA was similar between evaluations, but Halomax showed increased SDF at 4 h. The GSF group was similar between time points in both assays. Diluents did not affect SDF, irrespective of the assay. Halomax showed differences for BSF between time points, differently from SCSA. Linear regression did not show any correlation between assays. Conclusions: The use of Halomax should be encouraged for sperm DNA fragmentation analysis in horse frozen-thawed semen, particularly under field conditions.
Publication Date: 2018-05-08 PubMed ID: 29734417
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- Journal Article
Summary
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This research investigates the effects of semen cryopreservation on sperm DNA fragmentation in horses, using two different testing methods, and finds that Halomax might be more reliable for such tests, particularly in practical/field conditions.
Background
- The study focuses on sperm DNA fragmentation (SDF) in horses. More specifically, it investigates variability in SDF after semen cryopreservation in stallions that are regarded as either good or bad semen freezers (GSF or BSF).
- Semen cryopreservation refers to the freezing of sperm to preserve its viability for future use. This research is crucial as DNA damage during this process can adversely affect fertility, requiring ongoing analysis and surveillance.
Methods
- Thirteen stallions were categorized as GSF or BSF, and their ejaculates were cryopreserved using three different diluents.
- The sperm was assessed for SDF using two methods – the Sperm Chromatin Structure Assay (SCSA) and Halomax. These evaluations took place immediately after thawing and following a 4-hour thermoresistance test.
Results
- The BSF group exhibited consistent SDF results when evaluated using SCSA; however, the same group showed increased SDF after 4 hours when tested with Halomax. There were no significant changes in SDF in the GSF group as evaluated by both methods.
- No significant variance in SDF was observed from the utilization of different diluents in the testing process, regardless of the evaluation method used.
- Throughout the two time points, Halomax showed differences for BSF, unlike SCSA.
- There was no correlation observed between the two testing methods as per linear regression analysis.
Conclusions
- The research recommends the use of Halomax for analyzing sperm DNA fragmentation in frozen-thawed horse semen, especially in field conditions. This suggestion is based on the observation that Halomax proved more sensitive in detecting changes in SDF in BSF stallions over time, unlike SCSA.
Cite This Article
APA
Ferreira HN, Ferreira-Silva JC, Rocha JM, Farras MC, Calixto M, Moura MT, Alvarenga MA, Oliveira AL.
(2018).
Variable Inter-assay Estimation of Sperm DNA Fragmentation In Stallions Classified as Good and Bad Semen Freezers.
Cryo Letters, 39(1), 67-71.
Publication
Researcher Affiliations
- Laboratorio de Biotecnologias Reprodutivas, Departamento de Medicina Veterinaria (UFRPE), Recife-PE, Brasil.
- Laboratorio de Biotecnologias Reprodutivas, Departamento de Medicina Veterinaria (UFRPE), Recife-PE, Brasil.
- Laboratorio de Biotecnologias Reprodutivas, Departamento de Medicina Veterinaria (UFRPE), Recife-PE, Brasil.
- Laboratorio de Biotecnologias Reprodutivas, Departamento de Medicina Veterinaria (UFRPE), Recife-PE, Brasil.
- Faculdade Pio Decimo, Aracaju-SE, Brasil.
- Laboratorio de Biotecnologias Reprodutivas, Departamento de Medicina Veterinaria (UFRPE), Recife-PE, Brasil.
- Laboratorio de Reproduçao Animal, Departamento de Reproduçao Animal e Radiologia Veterinaria da Faculdade de Medicina Veterinaria e Zootecnia (UNESP), Botucatu-SP, Brasil.
- Laboratorio de Biotecnologias Reprodutivas, Departamento de Medicina Veterinaria (UFRPE), Recife-PE, Brasil. maloufrpe@gmail.com.
MeSH Terms
- Animals
- Biological Assay / methods
- Chromatin / metabolism
- Cryopreservation / veterinary
- DNA Damage
- DNA Fragmentation
- Horses / physiology
- Male
- Semen / metabolism
- Semen Analysis / veterinary
- Semen Preservation / veterinary
- Sperm Motility / physiology
- Spermatozoa / metabolism
Citations
This article has been cited 2 times.- Kumaresan A, Das Gupta M, Datta TK, Morrell JM. Sperm DNA Integrity and Male Fertility in Farm Animals: A Review. Front Vet Sci 2020;7:321.
- Boni R, Ruggiero R, De Luca F, Serritella ML, Di Palma T, Cecchini Gualandi S. Repeatability of Selected Parameters Related to Stallion Sperm Quality and Cryotolerance. Animals (Basel) 2025 Sep 26;15(19).
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