Pre-freezing equilibration for 22 h improves post-thaw sperm functions in cryopreserved ram semen by reducing cholesterol efflux |
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| Institution: | 1. Biotecnología de la Reproducción, Departamento de Producción Animal, Instituto Nacional de Tecnología Agropecuaria (INTA), Ruta 226 km 73.5, 7620, Balcarce, Argentina;2. Biología de Microorganismos y Gametas, Instituto de Investigaciones Biológicas CONICET, Universidad Nacional de Mar del Plata, Funes 3250, 7600, Mar del Plata, Argentina;3. Universidad Nacional de Mar del Plata, Ruta 226 km 73.5, 7620, Balcarce, Argentina;4. Laboratorio de Procesamiento de Imágenes ICYTE UNMDP – CONICET, Mar del Plata, Argentina;5. Reproductive and Developmental Biology Laboratory, National Institute of Environmental Health Sciences, Durham, NC, USA;1. Sezione di Clinica Ostetrica e Ginecologia, Dipartimento di Medicina Veterinaria, Università di Sassari, Via Vienna n.2, 07100 Sassari, Italy;2. Dipartimento di Medicina Clinica e Sperimentale, Università di Sassari, viale San Pietro n.10, 07100 Sassari, Italy;1. Laboratorio de Teriogenología “Dr. Héctor H. Morello”, Instituto de Biotecnología Agropecuaria del Comahue (IBAC). Facultad de Ciencias Agrarias, Universidad Nacional del Comahue, Cinco Saltos, Río Negro, Argentina;2. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Cinco Saltos, Río Negro, Argentina;3. Ministerio de Desarrollo Territorial, Gobierno de la Provincia del Neuquén, Neuquén, Argentina;1. Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Medicine, Universidad de La Frontera, Temuco, Chile;2. Department of Agricultural Sciences and Natural Resources, Faculty of Agriculture and Forestry Sciences, Universidad de La Frontera, Temuco, Chile;3. Department of Animal Production, Faculty of Agriculture and Forestry Sciences, Universidad de La Frontera, Temuco, Chile;4. School of Medical Technology, Faculty of Sciences, Universidad Mayor, Temuco, Chile;5. Center of Excellence in Translational Medicine, Universidad de La Frontera, Temuco, Chile;1. Dipartimento di Medicina Veterinaria, Sezione di Clinica Ostetrica e Ginecologia, Università di Sassari, Sassari, Italy;2. Dipartimento di Medicina Clinica e Sperimentale, Università di Sassari, Sassari, Italy;3. Laboratorio di Scienza dei Materiali e Nanotecnologie, D.A.D.U., Università di Sassari, CR-INSTM, Alghero, Italy |
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| Abstract: | Failure of cervical insemination with cryopreserved semen is hindering implementation of AI in sheep in field condition. Here the effect of equilibration time and catalase on post-thaw qualities of ram semen was investigated. Pooled semen was diluted (800 × 106 sperm mL?1) with a TES-Tris-fructose extender with 6% glycerol, 15% egg yolk and supplemented with 0, 50, 100 and 200 U mL?1 catalase and packaged into 0.25 mL straws. In experiment 1, straws were equilibrated at 5 °C either for 3 h in a cold cabinet (E3) or for 10 (E10) and 22 h (E22) inside a refrigerator. In experiment 2, all straws were equilibrated for 22 h inside refrigerator. Straws were frozen at ?25 °C min?1 up to ?125 °C using a cell freezer and finally plunged into liquid nitrogen. The post-thaw total and rapid motility were higher (P < 0.05) in E22 compared to E3 and E10. Sperm kinetics was comparable between E3 and E22, but lower in E10. Similarly, acrosome integrity, functional membrane integrity, percent high cholesterol (mCHO) and live-high mitochondrial membrane potential (MMP) were higher (P < 0.05) while live-high intracellular calcium and acrosome-reacted sperm were lower in E22 compared to E3 and E10. The percent rapid motile, high mCHO and live-high MMP were significantly (P < 0.05) lower in catalase-treated samples compared to the control, while the membrane integrity was comparable within the groups. In conclusion, pre-freezing equilibration for 22 h compared to 3 or 10 h resulted in higher post-thaw sperm functions while catalase had negative impact on cryopreservation of ram semen. |
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| Keywords: | Equilibration time Catalase Post-thaw quality Cryopreservation Semen Sheep |
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