Therio - Ai And Embryo Transfer

Front 1

Artificial Insemination
-main spp

Back 1

-bovine
-equine
-canine
-ovine

Front 2

Artificial insemination
-define

Back 2

-sperm is collected from a male and inserted by human action into the female reproductive tract to join with a naturally ovulated egg

Front 3

Sex-selected (sex-sorted) semen
-main spp

Back 3

-bovine
-equine

Front 4

Sex-selected semen
-define

Back 4

-sorting sperm cells into two containers, one containing predominantly X-bearing sperm and one containing predominantly Y-bearing sperm

Front 5

Embryo transfer
-main spp

Back 5

-bovine
-equine

Front 6

Embryo transfer
-define

Back 6

-natural service or AI is used to inseminate a femalt. The resulting embryo(s) are collected prior to the time of hatching from the zona pellucida and transferred to a different female for the rest of gestation

Front 7

In vitro fertilization
-main spp

Back 7

-experimental

Front 8

In vitro fertilization
-define

Back 8

-an egg is collected from a female and sperm is collected from a male and a single sperm cell is inserted into the egg in a lab environment
-embryo is cultured for several days and if it proceeds to the blastocyst stage, it is then transferred to the female

Front 9

Gamete Intra-fallopian transfer
-spp

Back 9

-experimental

Front 10

Gamete Intra-fallopian transfer
-define

Back 10

-egg is aspirated from a donor female's ovary prior to ovulation and then placed in the uterine tube of the recipient and breeding the recipient either naturally or AI

Front 11

Embryo splitting
-main spp

Back 11

-bovine
-equine

Front 12

Embryo splitting
-define

Back 12

-identical twins can be produced by splitting a day-6 embryo (blastocyst) and placing the embryo pieces into an empty zona pellucida and transferring them into recipient animals

Front 13

Cloning (Nuclear Transfer)
-main spp

Back 13

-bovine
-ovine
-canine
-equine

Front 14

Cloning (nuclear transfer)
-define

Back 14

-cells are removed from an embryonic blastocyst and implanted into an enucleated oocyte which is tranferred into a recipient female

Front 15

Cloning (somatic cell nuclear transfer)
-main spp

Back 15

-ovine
-canine
-bovine
-equine

Front 16

Cloning (somatic cell nuclear transfer)
-define

Back 16

-the nucleus is removed from an adult cell and re-programmed to allow differentiation into an entire organism when implanted into an enucleated egg which is transferred into a recipient female

Front 17

Artificial insemination
-semen that can be used

Back 17

-fresh
-chilled
-frozen

Front 18

AI
-advantages

Back 18

-genetic improvement: widespread use of males likely to produce valuable offspring or proven to have valuable offspring
-can use males anywhere in the world (animal health, easier to move than whole animals)
-don't have to handle as many males
-remove/reduce the risk of venereal disease

Front 19

Bull
-# ejaculate days per wk
-# straws per ejaculate
-# straws per week

Back 19

-2-3 ejaculated-days per wk
-150-500 straws per ejaculate
-300-1500 straws per week

Front 20

AI
-disadvantages

Back 20

-skill for estrus detection
-trained inseminator needed
-time and skill to commit to herd/animal reproduction
-increased cost per pregnancy

Front 21

AI
-semen collection

Back 21

-can be purchased from commercial suppliers and directly from male's owner
-can be collected on-site and used to mate many more female and decrease risk of injury via natural service
-can be collected by artificial vagina, electroejaculator, manual stimulation

Front 22

Spp semen that frequently frozen

Back 22

-bull
-dog
-stallion

Front 23

Semen Collection
-artificial vagina

Back 23

-males are trained to mount a dummy or teaser animal
-stallion: artificial vagina jacket is filled with warm water (temperature/pressure ejaculator)
-bull: artificial vagina does not need a warm water jacket (pressure ejaculator)

Front 24

Semen collection
-electroejaculation

Back 24

-used for males that are not trained to mount a dummy or teaser animal
-bulls, bucks, wildlife
-probe placed into rectum applies low voltage pulses to the pelvic nerved involved with erection and ejaculation

Front 25

Semen collection
-manual stimulation

Back 25

-dogs, boars, whales
-apply pressure and massage penis
-collect in a tube instead of an artificial vagina

Front 26

Sperm collection
-spermicidal material

Back 26

-latex
-plastic
-lubricants

-limit use and contact time

Front 27

Sperm cryopreservation
-adequate fertility of sperm post-thaw requires:

Back 27

-appropriate diluents/extenders
-appropriate sperm dilution
-appropriate cooling and thawing speed (cool fast, thaw fast)

Front 28

Sperm cryopreservation
-appropriate diluents

Back 28

-non-penetrating cryoprotectant (egg yolk, milk)
-penetrating cryoprotectant: protect from ice crystal formation (glycerol, ethylene glycol)
-buffer: prevent pH change (Tris, Test)
-sugar: provide energy (glucose, lactose)
-salt: provide proper osmolality (sodium citrate, citric acid)
-antibiotic: kill pathogens (penicillin, streptomycin)

Front 29

Appropriate sperm dilution

Back 29

-Bulls 20 million sperm per dose (50% live at thaw)

-can have less but must thaw and place sperm very well

Front 30

Sperm crypreservation
-cooling steps

Back 30

1) add extender ingredients and cool from 102F to 40F (refrigerator)
2) add cryoprotectants and package into straws (40F)
3) Freeze in liquid nitrogen vapor (-238F)
4) store in liquid nitrogen (-320F)

Front 31

How is semen stored?

Back 31

-a thermos-like tank with at least 2 inches of liquid nitrogen (-320F)

Front 32

Sperm cryopreservation
-thawing steps

Back 32

-best viability is thawed at 95F for 12 sec to 1 min
-once thawed, do not allow to cool
-don't allow to enter thaw-cool cycles

*faster thaw = better viability

Front 33

Most common limiting factor in AI success

Back 33

-accurate detection of estrus/ovulation

Front 34

Why would cryopreservation protocol for once species not work for another species?

Back 34

-sperm from different spp are different in size, shape, and lipid composition

Front 35

Canine
-ovulation occurs when

Back 35

-1-2 days after LH surge

*wait longer to breed after ovulation than other spp

Front 36

Canine
-AI with fresh semen

Back 36

-lives 5-6 days in the uterus
-breed 4-6 days after LH surge (3-5 days after ovulation)

Front 37

Canine
AI with chilled semen

Back 37

-lives approx 1-3 days after ejaculation
-when shipping, must plan ahead
-breed 4-5 days after LH surge (3-4 days after ovulation)

Front 38

Canine
-AI with frozen semen

Back 38

-short life span in uterus (12 hrs)
-need to inseminate intrauterine (poor motility for just putting intravaginal)
-breed 3-4 days after LH surge (2-3 days after ovulation)

Front 39

Canine
-methods for intrauterine delivery of frozen semen

Back 39

-surgical (laprotomy, laproscopy)
-transcervical catheterization

Front 40

Species where AI with frozen semen is just as fertile as natural service

Back 40

-bovine (bull)

Front 41

Bovine
-handling of frozen straws

Back 41

-make sure to keep unused straws frozen in canister
-lift tube up to neck of canister and remove straw with pair of tweezers
-thaw in 90-95F waterbath for 30-60 sec
-don't allow temp of straw to decrease

Front 42

Bovine
-where is the sperm deposited

Back 42

-past the cervix

*low dose
*reduced number of non-capacitated sperm

Front 43

Bovine
-Insemination technique

Back 43

-sleeved arm placed into rectum to guide cervix onto insemination gun
-insemination gun is inserted through the vulva and vagina then manipulated into the cervix
-once through the cervix depress the plunger in the body of the uterus (slowly)

Front 44

Bovine
-timing of insemination

Back 44

-12 hrs after first detected estrus
-want to be about 12 hrs before ovulation to allow time for capacitation

Front 45

Bovine
-egg lifespan

Back 45

-8-12 hrs

Front 46

Bovine
-sperm lifespan

Back 46

-40-48 hrs

Front 47

Bovine
-time of ovulation

Back 47

-24 hrs after start of standing estrus (10-14 hrs after end of estrus)

Front 48

Equine
-semen that does not need to be extended

Back 48

-fresh semen used immediately

Front 49

Equine
-semen that should be extended

Back 49

-fresh semen used 10 minutes or longer after collection

-store at room temp if used with 6-12 hrs (dark, air-tight container)
-remove seminal plasma to delay capacitation and extend lifespan
-maintain viability for > 12hrs cool to 40-45F over 203 hrs period

Front 50

Equine
-effect of cold shock on fresh semen

Back 50

-plasma membrane damage
-acrosome changes

Front 51

Equine
-shipping semen

Back 51

-ship in a container designed to maintain steady temperature

Front 52

Equine
-chilled semen viability

Back 52

-don't expect to be viable past 60-72 hrs (shipping concerns like canine)

Front 53

Equine
-when to inseminate with chilled semen

Back 53

-24 hrs prior to ovulation

Front 54

Equine
-how to induce ovulation

Back 54

-Deslorelin Acetate (GnRH agonist) to induce mares with >35 mm follicle (ovulation in 24-48 hrs)

*Sucromate Equine must be used

Front 55

Equine
-insemination timing with frozen semen

Back 55

-time within 12 hrs before up to 6 hrs after ovulation

*need frequent palpation or ultrasound

Front 56

Ovine
-using Ram Effect to aid synchronization

Back 56

-don't allow ewes to see or smell rams for at least 6 weeks prior to desired onset of cycling
-introduction of rams will induce ovulation in 3-4 days

Front 57

Ovine
-why is AI not typically done

Back 57

-difficult to detect estrus
-difficult to pass catheter through cervix

Front 58

Ovine
-types of AI

Back 58

-Cervical AI
-Transcervical AI
-Laparoscopic AI

Front 59

Ovine
-Cervical AI

Back 59

-speculum used to visualize the cervical os and fresh or chilled semen is deposited in the cervix

*don't use with frozen semen

Front 60

Ovine
-transcervical AI

Back 60

-special forceps used to pull the cervix into the vagina where another specialized scope is used to guide a catheter through the cervix

Front 61

Ovine
-Laparoscopic AI

Back 61

-surgical endoscope approach to deposit semen directly into uterus

*use with frozen semen

Front 62

Semen Sexing Technology
-why is it attractive to the dairy industry

Back 62

-desire for mostly heifers to serve as replacements

Front 63

Semen Sexing Technology
-why would it be attractive to beef producers

Back 63

-desire female calves from heifers calving for the first time (smaller calves)
-may desire male calves from mature cows (not worried about dystocia)

Front 64

Semen Sexing Technology
-how it's able to work

Back 64

Flow Cytometry
-Cells with X-chromosome contain slightly more DNA than sperm cells with Y-chromosome
-90% accuracy

*causes reduced sperm viability and longevity

Front 65

Semen Sexing Technology
-problems

Back 65

-slow process (6-10 straws/hr)
-need deep uterine insemination to compensate for reduced sperm dose
-more expensive
-fewer pregnancies than conventional semen
-can still have less than 79% of calves with desired sex

Front 66

Semen Sexing Technology
-companies

Back 66

-ABS
-Select Sires
-Genex

Front 67

Embryo Transfer
-benefit

Back 67

-allows use of genetically superior dams

Front 68

Embryo transfer
-superovulation

Back 68

-release of multiple eggs at a single estrus
-use multiple injections of FSH at 12 hr intervals

Front 69

Embryo transfer
-equine collection

Back 69

-collected from single ovulating mares

Front 70

Embryo transfer
-bovine superovulation

Back 70

-healthy and fertile donor female
-FSH injections every 12 hrs for 4 days mid diestrus
-PGF2a injected late in diestrus (4th day of FSH)
-use ultrasound to determine # of ovulations

Front 71

Embryo Transfer
-how to breed a superovulating donor

Back 71

-lots of eggs = lots of sperm
-2 straws every 12 hrs
-semen proven to be fertile post-thaw

Front 72

Embryo transfer
-collecting embryos

Back 72

-bovine = 7 days post estrus
-equine 7-8 days after ovulation (day 6 if plan on freezing)

-when embryos not hatched from zona pellucida
-give an epidural to prevent straining
-fluid used = phosphate buffered saline + bovine serum
-high quality embryos collected for immediate transfer or frozen

Front 73

Embryo transfer
-embryo quality grades

Back 73

-Grade 1 = excellent to poor
-Grade 2 = fair
-Grade 3 - poor
-Grade 4 = dead or degenerating

*can transfer grades 1-3
best success with grades 1 & 2

Front 74

Embryo transfer
-embryos that should be frozen

Back 74

-grade 1

Front 75

Embryo transfer
-recipients

Back 75

-healthy and fertile
-have 8 recipients if transferring fresh embryos (generally get 6-8)
-synchronize recipients so they are on the same day of the estrous cycle as the donor (Day 7 - bovine)
-mare recipients should have ovulated the day before or up to 3 days after the donor

Front 76

Embryo transfer
-transfer

Back 76

-deposit in horn on same side of CL

Front 77

Embryo transfer
-embryo quality criteria

Back 77

-regularity of embryo shape
-compactness of blastomeres
-variation in cell size
-color and texture of cytoplasm
-overall diameter of embryo
-presence of extruded cells
-regularity of zona pellucida
-presence of vesicles in cytoplasm