Abstract
The aim of this study was to establish and validate a reliable and efficient protocol for the recovery and cryopreservation of epididymal spermatozoa used for in vitro fertilization, using bulls of two different age classes. Testicles from 26 (37-51 weeks old, group 1) and 19 (52-115 weeks old, group 2) Danish Holstein bulls were collected after slaughter and stored at 5°C. After 0, 24 or 48 h, epididymides were isolated and spermatozoa collected. Assessments included spermatozoal motility, viability and morphology before and after cryopreservation and in vitro embryo production. Results showed that live spermatozoa can be collected from epididymides of bulls after their death. Storage of the testicles at 5°C for 24 h followed by cryopreservation of recovered epididymal spermatozoa resulted in 21% (group 1) and 31% (group 2) blastocysts produced in vitro. These results illustrate that epididymal spermatozoa recovered from testicles kept in specific conditions can be used to preserve genetic material from endangered and threatened species or populations in nature as well as in domestic and zoo animals.
Highlights
The ability to preserve genetic material is an important tool in conserving genetic variation in endangered populations and species
One testicle was processed at this time, whereas the other testicle was stored at 5°C in a plastic bag and processed after 24 h (T24; 14 from group 1 and 10 from group 2) or 48 h (T48; 12 from group 1 and nine from group 2)
The protocol used in this study was efficient in generating viable and motile epididymal bull spermatozoa, which could be cryopreserved and subsequently used successfully for in vitro fertilization (IVF) with higher developmental rates than described previously
Summary
The ability to preserve genetic material is an important tool in conserving genetic variation in endangered populations and species. Material in animal biobanks is useful for various types of research, such as cryobiology, reduction of inbreeding, genomic selection studies, assessment of genetic distances and disease genetics (Blackburn, 2012; Groeneveld et al, 2016). When working with domesticated animals, zoo animals and wildlife populations of threatened animals, the number of breeding males is often limited to a few superior animals. Suitable protocols for recovery of spermatozoa after the animal’s death will enable us to exploit and preserve genetic material by cryopreservation of spermatozoa or in vitro production of embryos
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