Single-Layer Colloid Centrifugation as a Method to Process Urine-Contaminated Stallion Semen After Freezing-Thawing |
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| Institution: | 1. Department of Veterinary Clinical Medicine, College Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL;2. Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, Athens, GA;3. Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY;1. Department of Animal Sciences, Colorado State University, Fort Collins, CO;2. Department of Clinical Sciences, Colorado State University, Fort Collins, CO;3. Department of Biomedical Sciences, Colorado State University, Fort Collins, CO;1. UNIPAC- Faculdade Presidente Antônio Carlos, Uberlândia, Minas Gerais, Brazil;2. UNESP-Universidade Estadual Paulista, Campus de Botucatu, Department of Veterinary Clinical Science, School of Veterinary Medicine and Animal Science, Botucatu, São Paulo, Brazil;3. UNESP-Universidade Estadual Paulista, Campus de Botucatu, Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science, Botucatu, São Paulo, Brazil;4. UNESP- Universidade Estadual Paulista, Campus de Botucatu, Department of Anatomy, Biosciences Institute, Botucatu, São Paulo, Brazil;5. UNESP- Universidade Estadual Paulista, Campus de Botucatu, Department of Biostatistics, Biosciences Institute, Botucatu, São Paulo, Brazil;1. Department of Animal Genetics, Breeding & Reproduction, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Liaoning Shenyang, PR China;2. Department of Clinical Veterinary Medicine, College of Veterinary Medicine, China Agricultural University, Beijing, PR China;1. Chief Veterinary Officer of the Veterinary Unit, Zaragoza, Spain;2. Department of Animal Pathology, Faculty of Veterinary Medicine, Instituto Agroalimentario de Aragón IA2 (Universidad de Zaragoza-CITA), Zaragoza, Spain;3. Chief Veterinary Officer of the Veterinary Unit, Jerez de la Frontera, Cádiz, Spain;1. Department of Emergency and Organs Transplantation, Italy;2. Department of Veterinary Medicine University of Bari-Aldo Moro, Italy;3. Interdisciplinary Department of Medicine, University of Bari-Aldo Moro, Italy;4. Prevention Department-Veterinary service, area A of the Local Healthcare (ASL), Bari, Italy;1. Division of Reproduction, Department of Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Box 7054, 75007 Uppsala, Sweden;2. Department of Surgery and Theriogenology, College of Veterinary Medicine, University of Mosul, Mosul, Iraq |
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| Abstract: | Urospermia is a major ejaculatory dysfunction affecting stallions. It has been thought that urine-contaminated semen should not be cryopreserved; however, on select cases, urine contamination of semen cannot be avoided. A recent study suggested that urospermic semen can be cryopreserved after cushion centrifugation and extension. Thus, this study aimed to assess the use of single-layer colloid centrifugation (SLC) to process frozen-thawed urine-contaminated stallion semen. Raw ejaculates (n = 55) from eight stallions were split into three groups: no urine, low (20%), or high (50%) urine contamination. Semen was extended 1:1, cushion-centrifuged, and resuspended at 200 million sperm/mL in BotuCrio. Resuspended semen was loaded in 0.5 mL straws and cryopreserved in liquid nitrogen. Samples were thawed (37°C for 30 seconds) and processed by SLC (400 g/30 minutes). Percentages of total motility (TM) and progressive motility (PM) were assessed with computer-assisted semen analyzer. Sperm viability (%VIAB) and yield were assessed with a NucleoCounter before and after gradient centrifugation. Data were analyzed with two-way ANOVA and Tukey’s test. The motility parameters TM before SLC (control: 35 ± 2; low: 33 ± 0.7; high: 22 ± 1.8) after SLC (control: 51 ± 3.6; low: 42 ± 2.2; high: 25 ± 2.8) and PM before SLC (control: 24 ± 1.8; low: 21 ± 1.14; high: 12 ± 1.5) and after SLC (control: 40.3 ± 3.2; low: 31 ± 3.9; high: 14 ± 2) significantly decreased with increasing urine contamination. Urine contamination marginally reduced (P < .05) sperm viability after cryopreservation before SLC (control: 45 ± 0.7; low: 27 ± 0.2; high: 27 ± 0.3) and after SLC (control: 54 ± 0.5; low: 49 ± 0.7; high: 38 ± 0.6). Recovery rates of sperm after centrifugation were not significantly different between groups. In conclusion, urine contamination affects sperm motility parameters in a dose-dependent manner. Post-thaw SLC selected sperm with higher motility and viability in control and low groups but only selected sperm with higher viability in the high group. |
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| Keywords: | Horse Urospermia Centrifugation Cryopreservation |
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