1. Freezing chicken semen: Influence of base medium osmolality, cryoprotectants, cryoprotectant concentration, and cooling rate on post-thaw sperm survival
- Author
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H. Woelders, A.A.C. de Wit, B. Engel, B. Hulsegge, I. Grasseau, E. Blesbois, B. Bernal, J. Santiago-Moreno, Ministry of Agriculture, Nature and Food Quality (The Netherlands), Engel, B., Hulsegge, B., Grasseau, I. 0000-0002-2692-2340], Blesbois, E., Bernal, B., Santiago-Moreno, J., Wageningen University and Research [Wageningen] (WUR), Biometris, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria = National Institute for Agricultural and Food Research and Technology (INIA), and European Project: 677353
- Subjects
Male ,Animal Breeding and Genomics ,Wiskundige en Statistische Methoden - Biometris ,General Biochemistry, Genetics and Molecular Biology ,Cryoprotective Agents ,Semen ,Acetamides ,Freezing ,Animals ,Life Science ,Fokkerij en Genomica ,Fokkerij & Genomica ,Mathematical and Statistical Methods - Biometris ,Cryopreservation ,Dimethylacetamide ,Cooling rate ,Osmolar Concentration ,Freezing medium ,CPA concentration ,[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology ,Dimethylformamide ,General Medicine ,Propylene Glycol ,Spermatozoa ,CPA ,Extender ,Chicken semen ,Sperm Motility ,WIAS ,General Agricultural and Biological Sciences ,Chickens ,Semen Preservation ,Animal Breeding & Genomics - Abstract
11 Pág. Departamento de Reproducción animal, A classical chicken semen diluent (Lake's 7.1 diluent) was modified to have lowered osmolalities (ranging from 290 to 410 mOsm/kg). The modified medium with physiological osmolality of 325 mOsm/kg allowed cold storage of fresh semen for several days with very little loss of membrane integrity and motility, while high osmolalities inhibited motility. This modified medium was then used as base for freezing medium to test effects of the type and concentration of cryoprotective agent (CPA), and the cooling rate (CR). A number of CPAs (methylformamide, methylacetamide, dimethylformamide (DMF), dimethylacetamide (DMA), diethylformamide, and propylene glycol) were first compared by freezing semen with 0.6 mol/l of the respective CPA at a cooling rate of 250 °C/min. Post-thaw motility and membrane integrity were highest with DMA and DMF. Finally, in more detailed factorial experiments, semen from individual cocks or pooled semen was frozen using CRs of 4, 50, 250, and 440 °C/min and DMA concentrations ([DMA]) of 0.4, 0.6, 1.0, and 1.5 mol/l. Straws from each semen sample x treatment combination were divided for semen assessment at three different research groups for sperm motility, membrane integrity, kinked tails, and DNA fragmentation, using microscopy, computer assisted motility analysis, and flow cytometry. There were clear effects of both CR and [DMA] and their interaction. CRs 50 and 250 °C/min gave best post-thaw sperm performance. Higher DMA concentrations gave better post-thaw membrane integrity, but concentrations above 1.0 mol/l can decrease sperm velocity or even inhibit sperm motility. Therefore [DMA] may best be 0.6-1.0 mol/l at a CR of 50-250 °C/min., Funding from the Dutch Ministry of Agriculture, Nature and Food Quality (project KB-Dier, WOT-03-436-077) and from European Union's Horizon 2020 Research and Innovation Programme (project IMAGE, grant agreement no 677353) are gratefully acknowledged.
- Published
- 2022
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