Semen Extension and Cryopreservation

Question 1

Dilute semen in ______ to maintain sperm viability

Answer 1

Buffered extenders

Question 2

_______ varies per species

Answer 2

Semen extension

Question 3

Extenders differ in composition depending upon the _____,_____,______and ______

Answer 3

species,
intended use,
storage temperature,
the desired duration of storage

Question 4

Buffers in extenders modulate the change in extender pH due to _____________

Answer 4

metabolic products of stored spermatozoa

Question 5

Extenders contain freezing point depressing compounds, which ________,_________, and _________

Answer 5

inhibit ice formation,
dehydrate cells,
and protect cellular structures

Question 5

Extenders maintain sperm viability at ________and control bacterial growth

Answer 5

20 deg. Celsius

Question 6

Bovine: (Procedure 1)
To pass for processing, semen samples should have an initial minimum ________progressive
motility and ______ normal morphology

Answer 6

minimum 75% progressive
motility and,
70% or greater normal morphology

Question 7

Bovine: (Procedure 2)
Semen aliquot is diluted 100-fold in __________, and ______are added for pathogen control

Answer 7

saline solution and atbcs are added for pathogen control

Question 8

Bovine: (Procedure 3)
__________ is incubated prior to extension since the activity of _____is hindered in the presence of milk or egg yolk in semen extenders

Answer 8

Atbc-semen mixture is incubated prior to extension since the activity of atbcs is hindered in the presence of milk or egg yolk in semen extenders

Question 9

Bovine: (Procedure 4)
Semen is then added to _________, then placed in an __________
placed into a 5 deg. celsius cold cabinet, cooled for several hours

Answer 9

Semen is then added to isothermal extender, then placed in an isothermal water jacket
placed into a 5 deg. Celsius cold cabinet, cooled for several hours

Question 10

Bovine: (Procedure 6)
Semen is then diluted to final desired concentration with ________, packaged
in _______ plastic french straws, sealed, and frozen for 10 minutes in _________ at a starting temperature of ________

Answer 10

Semen is then diluted to final desired concentration with glycerolated extender, packaged in 0.5 mL plastic french straws, sealed, and frozen for 10 minutes in static nitrogen vapor at a starting temperature of -160 deg. celsius

Question 11

Bovine: (Procedure 5)
After initial cooling, samples are ___________________

Answer 11

removed from water jackets and maintained at 5 deg. celsius

Question 12

Bovine: (Procedure 7)
After freezing, the straws are immersed into _______ until the time of _______, through placing it into a ________water bath for 30-60 seconds

Answer 12

After freezing, the straws are immersed into liquid nitrogen until the time of thawing, through placing it into a 35 deg. water bath for 30-60 seconds

Question 13

Bovine: (Procedure 9)
Post-thaw analysis minimum standards:
1. Motility
2. Spermatozoal progressive linear motility
3. Intact acrosomes

Answer 13

Post-thaw analysis minimum standards:
1. Motility: 25% (0hrs)- 15% (2hrs)
2. Spermatozoal progressive linear motility: 3-2
3. Intact acrosomes: 60-40%

Question 14

Bovine: (Procedure 8)
All _________are performed at 35 deg. celsius

Answer 14

post-thaw evaluations

Question 15

Equine (Procedure 1):
Stallion semen may be diluted with _____ or _____, then used immediately or cooled to ____and stored for AI of mares within _________

Answer 15

Stallion semen may be diluted with egg-yolk buffer or milk solids-glucose extenders, then used immediately or cooled to 5°C and stored for AI of mares within 24 to 48 hours of ovulation

Question 16

Equine (Procedure 2):
Semen may be centrifuged to_________

Answer 16

remove seminal plasma to concentrate sample

Question 17

Equine (Procedure 3):
Soft pellet is then extended at 20°C in a_____________ medium containing ______

Answer 17

sodium-citrate-EDTA lactose-egg yolk medium containing cryoprotectant, glycerol

Question 18

Equine (Procedure 4):
Final extended semen should have a concentration sufficient to provide ________, normal cells after thawing

Answer 18

at least 200 x 106 motile

Question 19

Equine (Procedure 5):
Semen is then packaged into plastic straws ranging from __________

Answer 19

0.5 mL to 5.0 mL

Question 20

Equine (Procedure 6):
Freezing is accomplished in a ______ or ______ nitrogen vapor unit, then immediately immersed in ___________

Answer 20

Freezing is accomplished in a passive or mechanically forced nitrogen vapor unit, then immediately immersed in liquid nitrogen for storage at -196°C

Question 21

Ovine (Procedure 1):
Ram semen can be extended for fresh insemination, _____________ or ________________

Answer 21

cooled for short term storage (24-48hours) or stored frozen for long-term preservation

Question 22

Ovine (Procedure 2):
Extender

Answer 22

tris-glucose (1 part semen / 2parts extender)

Question 23

Ovine (Procedure 3):
Thawing

Answer 23

place straws in 35°C water for at least 30 seconds

Question 24

Caprine (Procedure 1): Goats inseminated with ___________

Answer 24

fresh extended, cooled, or frozen semen

Question 25

Caprine (Procedure 2): Extender

Answer 25

milk or tris-citrate-egg yolk (4-5%) extenders, glycerol

Question 26

Caprine (Procedure 3): Initial colling is to _____ in _____

Answer 26

Initial colling is to 5°C in non-glycerolated extender

Question 27

Caprine (Procedure 4): _________ is then added slowly and the mixture us allowed to equilibrate over _____ before freezing in nitrogen vapor and storage at ______

Answer 27

Glycerolated portion is then added slowly and the mixture us allowed to equilibrate over 2 hours before freezing in nitrogen vapor and storage at -196°C

Question 28

Caprine (Procedure 5): Thawing

Answer 28

35°C, 30 seconds before insemination

Question 29

Porcine (Procedure 1): Collected ejaculate is added to a ______and cooled to _______

Answer 29

Collected ejaculate is added to a predilution extender and cooled to 18°C over 4 hours

Question 30

Porcine (Procedure 2): Semen centrifuged for ______ at _____and the pellet resuspended in ________________

Answer 30

Semen centrifuged for 10 minutes at 18°C and the pellet resuspended in a cooling diluter containing egg yolk

Question 31

Porcine (Procedure 3): Semen cooled from _________

Answer 31

18°C to 5°C

Question 32

Porcine (Procedure 4): Before freezing, semen is diluted to a final concentration of __________ in a ________, then frozen in 5 mL plastic tubes

Answer 32

1.2 x 109 spermatozoa/mL in a deep-freeze diluter at 4% glycerol, then frozen in 5 mL plastic tubes

Question 33

Porcine (Procedure 5): Thawing

Answer 33

52°C for 50 seconds