extenders commonly used in the breeding industry of domestic
species include Minitube and IMV Technologies.
3. Approximately ~150 mL will be recovered from 10 mediumsized eggs.
4. If the obtained fraction is not “clear,” the centrifugation duration can be increased for example, to 30 min, or the obtained
fraction can be subjected to a second centrifugation round
using the same conditions.
5. Instead of egg yolk, low-density proteins (LDLs) or liposomes
of nonanimal origin can be used.
6. Antibiotic types and concentrations should be added to cryopreserved semen as recommended by the Terrestrial Animal
Health Code of the World Organization for Animal Health,
and further international regulations (e.g., European Council
Directives), and reported on the accompanying transport certificate. A combination of antibiotics need to be used with a
bactericidal activity at least equivalent to gentamicin (250 μg/
mL), tylosin (50 μg/mL), and lincomycin-spectinomycin
(150/300 μg/mL); penicillin (500 IU/mL), streptomycin
(500 μg/mL), and lincomycin-spectinomycin (150/300 μg/
mL); or amikacin (75 μg/mL) and dibekacin (25 μg/mL).
7. When the osmolality of a solution is too high, add water while
taking into account how much the sample should be diluted.
When the osmolality is too low, calculate the number of
osmoles missing (i.e., difference from, for example, 300 or
330). Take into account the solution volume: the recorded
osmolality represents the number of osmoles per kg water or
L. In case of INRA-82, the major component is glucose; thus,
glucose can be added to reach the desired osmolality. Glucose
does not dissociate in solution; thus, the number of osmoles
missing in the solution equals the number of moles to be
added. If correcting an osmolality by adding NaCl, e.g., for a
saline solution, remember it dissociates in solution and the
number of osmoles missing should be divided by 2 for obtaining the number of moles to be added. Calculate the amount of
grams to be added to the solution volume by multiplying the
number of moles needed with the molecular weight of the
compound.
8. Skim milk used in extenders should be heated to ~90
C for
10 min in order to inactivate the lactenin which is toxic to
stallion sperm. Skim milk can be replaced with defined proteins
such as phosphocaseinates.
9. Equex (STM) and Orvus ES paste contain the detergent/surfactant sodium dodecyl sulfate. Addition likely results in solubilizing egg yolk lipids, which has been implicated in increasing
protective properties.
Cryopreservation of Sperm from Domestic Livestock
375
species include Minitube and IMV Technologies.
3. Approximately ~150 mL will be recovered from 10 mediumsized eggs.
4. If the obtained fraction is not “clear,” the centrifugation duration can be increased for example, to 30 min, or the obtained
fraction can be subjected to a second centrifugation round
using the same conditions.
5. Instead of egg yolk, low-density proteins (LDLs) or liposomes
of nonanimal origin can be used.
6. Antibiotic types and concentrations should be added to cryopreserved semen as recommended by the Terrestrial Animal
Health Code of the World Organization for Animal Health,
and further international regulations (e.g., European Council
Directives), and reported on the accompanying transport certificate. A combination of antibiotics need to be used with a
bactericidal activity at least equivalent to gentamicin (250 μg/
mL), tylosin (50 μg/mL), and lincomycin-spectinomycin
(150/300 μg/mL); penicillin (500 IU/mL), streptomycin
(500 μg/mL), and lincomycin-spectinomycin (150/300 μg/
mL); or amikacin (75 μg/mL) and dibekacin (25 μg/mL).
7. When the osmolality of a solution is too high, add water while
taking into account how much the sample should be diluted.
When the osmolality is too low, calculate the number of
osmoles missing (i.e., difference from, for example, 300 or
330). Take into account the solution volume: the recorded
osmolality represents the number of osmoles per kg water or
L. In case of INRA-82, the major component is glucose; thus,
glucose can be added to reach the desired osmolality. Glucose
does not dissociate in solution; thus, the number of osmoles
missing in the solution equals the number of moles to be
added. If correcting an osmolality by adding NaCl, e.g., for a
saline solution, remember it dissociates in solution and the
number of osmoles missing should be divided by 2 for obtaining the number of moles to be added. Calculate the amount of
grams to be added to the solution volume by multiplying the
number of moles needed with the molecular weight of the
compound.
8. Skim milk used in extenders should be heated to ~90
C for
10 min in order to inactivate the lactenin which is toxic to
stallion sperm. Skim milk can be replaced with defined proteins
such as phosphocaseinates.
9. Equex (STM) and Orvus ES paste contain the detergent/surfactant sodium dodecyl sulfate. Addition likely results in solubilizing egg yolk lipids, which has been implicated in increasing
protective properties.
Cryopreservation of Sperm from Domestic Livestock
375