Chapter 16
Cryopreservation of Avian Semen
Henri Woelders
Abstract
Cryopreservation protocols for semen exist for bird species used in animal production, fancy and hobby
species, and wild bird species. Freezing of bird oocytes or embryos is not possible. Cryopreservation of avian
semen is used for preserving (genetic diversity of) endangered species or breeds. Freezing semen can also be
used in the breeding industry for maintaining breeding lines, as a cost-effective alternative to holding live
birds. Success and efficiency of cryopreservation of bird semen differs among species and breeds or selection
lines. This chapter describes important variables of methods for collecting, diluting, cold storage, and
freezing and thawing of bird semen, notably the medium composition, cryoprotectant used and its
concentration, cooling rate, freezing method, and warming method. Media and methods are described
for freezing semen using either glycerol or DMA as cryoprotectant, which both are known in chicken and a
number of other bird species to render adequate post-thaw fertility rates.
Key words Avian, Poultry, Semen, Cryopreservation, Extender, Insemination
1 Introduction
1.1 Historical
Perspective
Poultry spermatozoa were, in fact, the first spermatozoa and arguably the first vertebrate cells to be frozen and thawed successfully
with significant post-thaw viability, which was made possible by the
serendipitous discovery of the cryoprotective property of glycerol
[1]. The fertilizing ability of the frozen fowl semen was not immediately evident due to the contraceptive effect of glycerol. But after
introducing the removal of the glycerol by dialysis, the chicks that
were then obtained in 1951 were the first vertebrates in the world
to be produced from eggs fertilized with sperm preserved by freezing [2, 3]. The cryoprotective action of glycerol and other neutral
solutes was explained by Lovelock [4] as being due to the ability of
these compounds to permeate through the cell membrane and to
reduce the rise of intra- and extracellular electrolyte concentration
during freezing. In fact, a great number of similar membranepermeating compounds can be effectively used as CPA for freezing
live cells.
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_16, © Springer Science+Business Media, LLC, part of Springer Nature 2021
379
Cryopreservation of Avian Semen
Henri Woelders
Abstract
Cryopreservation protocols for semen exist for bird species used in animal production, fancy and hobby
species, and wild bird species. Freezing of bird oocytes or embryos is not possible. Cryopreservation of avian
semen is used for preserving (genetic diversity of) endangered species or breeds. Freezing semen can also be
used in the breeding industry for maintaining breeding lines, as a cost-effective alternative to holding live
birds. Success and efficiency of cryopreservation of bird semen differs among species and breeds or selection
lines. This chapter describes important variables of methods for collecting, diluting, cold storage, and
freezing and thawing of bird semen, notably the medium composition, cryoprotectant used and its
concentration, cooling rate, freezing method, and warming method. Media and methods are described
for freezing semen using either glycerol or DMA as cryoprotectant, which both are known in chicken and a
number of other bird species to render adequate post-thaw fertility rates.
Key words Avian, Poultry, Semen, Cryopreservation, Extender, Insemination
1 Introduction
1.1 Historical
Perspective
Poultry spermatozoa were, in fact, the first spermatozoa and arguably the first vertebrate cells to be frozen and thawed successfully
with significant post-thaw viability, which was made possible by the
serendipitous discovery of the cryoprotective property of glycerol
[1]. The fertilizing ability of the frozen fowl semen was not immediately evident due to the contraceptive effect of glycerol. But after
introducing the removal of the glycerol by dialysis, the chicks that
were then obtained in 1951 were the first vertebrates in the world
to be produced from eggs fertilized with sperm preserved by freezing [2, 3]. The cryoprotective action of glycerol and other neutral
solutes was explained by Lovelock [4] as being due to the ability of
these compounds to permeate through the cell membrane and to
reduce the rise of intra- and extracellular electrolyte concentration
during freezing. In fact, a great number of similar membranepermeating compounds can be effectively used as CPA for freezing
live cells.
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_16, © Springer Science+Business Media, LLC, part of Springer Nature 2021
379