Bovine Repro Final Flashcards
(187 cards)
historical procedure for embryo transfer
full surgery with vets/techs/anesthesiologists etc. there is a middle incision and flank incision to flush into large container (container can hold lot of fluid so there is a lot to sort through to find) –> later made filter
then to transfer gun needs to make it into uterine horn so embryo can shot out front or sides and need to make sure it is in the side of the horn with CL (use palpation)
what is the embryo transfer filter
plastic embryo collection filter with 70 micron stainless steel filter
lid with in-flow connection, vent cup lid etc
embryo transfer (finding oocyte)
need stereomicroscope (10-50x suitable for bovine repro embryology
factors affecting success for ET
- time in holding medium: at 2 - 7hrs can have almost 6,000 transfers with 73% preg vs 19 - 24hrs can have 150 transfers and 72% preg
- embryo age day of flush: best to flush cows day 7 but 6 and 8 have no statistical difference
- using invivo vs invitro
- fresh > frozen
scales for grading embryos
IETS 1 - 4 (international embryo transfer society)
1 = excellent/good 2 = fair 3 = poor 4 = degenerate
want to have 1 or 2 (2 has 63% pr vs 3 has 46% pr)
embryo transfer recipient-donor estrus synchrony
increased percent pregnant at 24hrs on both ends (plus/minus) when being synched
sever decrease if using frozen embryos 24hrs after synching
embryo transfer on-farm factors (4)
management
synchronization of donors
nutrition
seasonal effects
invivo vs invitro (ET)
invivo: day 0 is the time you detect first standing heat, embryos are less advanced, should be within 24hr synch
invitro: day 0 is considered maturation, embryos have blastocysts and expanded blastocysts
vitro uses more matured egg
new tools
new/greatly improved tools appear constantly, about every 7 years truly novel tool occur which revolutionize science and application
1/2 noble prizes in physiology or medicine concern new tools
superovulation
neew tech to superovulate cows and collect much more embryos per flush, about 60% use FSH about twice daily for 3 to 4 days
cryopreservation of embryos
allow movement of valuable genetics, no need for large recipient herd, preserve genetics from deceased animals, can be coupled with other technologies (embryo biopsy)
gly (glycerol) eg (??)
revolutionary tools (5) ET
transgenic technology, stem cell biology, somatic cell nuclear transplantation, polymerase chain reaction, fertilization by sperm injection
IVF history
1959 first to be done in mammal being a rabbit
1981 first calf from IVF
1988 1st repeatable OPU (ovum pick up) protocol
1990s production of calves from IVF commercially established internationally
IVD (in vivo derived) trends decrease and IVP (in vitro produced) trends increase
IVP vs IVD
IVD = in vivo derived
IVP = in vitro produced
IVP > IVD trending
source of oocytes
OPU = ovum pick-up or transvaginal oocyte aspiration
slaughterhouse/deceased animals
why surge in IVF embryo production?
advances in collection, handling, processing, storage, transport, transfer
improved equipment for OPU and improvement in super stimulation protocols
more in dairy breeds but increasing overall, can leave embryo out for 24 hr and won’t see decrease in preg rate vs oocyte need to be more careful with temp
collecting oocyte process
oocytes are sensitive to temperature, move oocytes into an incubator as quickly as possible, grading for oocytes
storage = slow rate freezing?, vitrification = freezing so quickly liquid doesn’t form ice but instead glass-like solid, improvement in media
transport = portable incubators, maturation during shipment, shipping media does not require equilibrium
steps in in vitro embryo poduction (3)
- maturation
- fertilization
- embryo culture
fertilization: AI 2 mil put in for semen, usually get about 100 sperm trying to fertilize egg, capacitation that change sperm plasma membrane, chose frozen over fresh for invitro
maturation in vitro vs in vivo
in vivo = final maturation due to LH surge in vivo, cAMP maintains meiotic arrest by inhibiting PDE3, LH surge -> decrease cAMP -> releases inhibition of PDE3 -> resumption of meiosis
invitro = final maturation due to removal from an inhibitory environment, removal from inhibitory environment initiates maturation
capacitation
requires changes in the sperm plasma membrane so sperm is able to penetrate and fertilize the egg, sperm aquire the ability to fertilize oocytes (acromse reaction, hyperactivated motility) cryopreservation of sperm facilities capacitation
methods of inducing capacitation in vitro
heparin = binds to proteins that stimulate loss of membrane cholesterol and phospholipids increase Ca acrosome
caffeine = increase cAMP
BSA = removes cholesterol from membrane
freeze/thaw sperm = destabilizes membrane
ICSI
intracytoplasmic sperm injection\
dont usually do/does work well for bovine but will use when sperm are less mobile or don’t swim well
culture systems ET media
1 vs 2 vs 3 step media
composition of media is important: energy substrates, buffers, amino acids, antioxidants, macromolecules, osmolytes
why use IVF
females that process abnormalities in their reproductive tracts, terminal females, improves efficiency of sperm if semen is rare or expensive, can aspirate oocytes during the first 90 days of pregnancy