Lab Final Flashcards
Reproductive technologies
Semen sexing
IVP of embryos
Transgenic animal production
Gene mapping
Marker assisted selection
Transgenic animal production examples
Micro injection
Blastocyst injection
Nuclear transfer
Propagate superior livestock animals
Cloning
Remove or add a gene to improve a breed of livestock
Transgenics
Select animals for specific traits very early in life
Coat color in cattle
Production of proteins which are essential to human life
Milk
Blood
Urine
Alter specific genes in specific locations to attain a desirable result
Transgenics
What chromosome gives off more light emission because it has more dye and more DNA
X chromosome
What chromosome has an extra short arm
X chromosome
What determines sex
Sperm
What excites vital dye in semen sexing
Laser
Why does the X chromosome have more dna
Extra short arm
Sperm run through system ______
Individually
Sperm is given a charge based on
Light intensity
Pass magnetic field in semen sexing
Sorting
Dead or damaged sperm not selected
Purifying samples
System that has a 95% or better success sorting
Flow cytometric analysis of sperm
Ovaries isolate
Oocytes
Step 1 of in vitro fertilization
Synchronization of recipients with donor
Step 2 of in vitro fertilization
Superovulation of donor
Multiple treatments of porcine FSH promotes more
Follicle development
8 CLs mean the donor is
Super ovulated
Step 3 or in vitro fertilization
Inseminate donor with semen from genetically superior bull
Work AI rod through the ______ by ______ ________
Cervix
Rectal palpation
Once rod is beyond the cervix in the uterine body then you
Deposit semen
Step 4 of in vitro fertilization
Recovery and identification of viable embryos
Collect/flush the embryos in the
Uterine horn
Step 5 of in vitro fertilization
Transfer of viable embryos into synchronized recipients
Cattle: Want the recipient to be a day _____, a day ______, or
Ahead
Behind
Exact synchrony with donor
Pig: want recipients to have _____ ______ or be a little _______
Exact synchrony
Behind
Cow transfer of viable embryos into synchronized recipients days
5, 6, or 7
Pig transfer of viable embryos into synchronized recipients days
5 or 6
Long ultrasound probe inserted into reproductive tract through vulva into vagina
Transvaginal aspiration
Always better to transfer ______ than to freeze
Immediately
Provides suction and holds embryo in place
Holding pipette
Snap on end-injection needle —> into pro nucleus
Glass pipette
Embryos are not designed to be centrifuged at ____ speeds
High
Efficiency of trans gene integration following micro injection
10% success rate mouse embryos
Livestock even less efficient
Blastocyst injection of embryonic stem cells occurs at a much ____ stage in development
Later
What houses the developing fetus during gestation
Placenta
Cells can either be ______ ____ ____ or _______
Inner cell mass
Trophoblast
Fluid-filled area
Blastocoel
Forms embryo
Inner cell mass
Forms placenta
Trophoblast
Day 6-7 embryo development
Compact morula
Day 7-8 embryo development
Early blastocyst
Day 7-9 embryo development
Blastocyst
Day 8-10 embryo development
Expanded blastocyst
Day 9-11 embryo development
Hatching blastocyst
Trophoblast cells die off means a
Pure population of inner cell mass cells (embryonic stem cells)
Embryonic stem cells can utilize
Gene targeting
Can add a gene, replace a gene, or remove a gene in a very specific location
Gene targeting
Gene targeting needs _____ of embryonic stem cells
Millions
Gene is swapped with genetic alteration
Homologous recombination
Select cells to isolate only those that were able to take out that genetic alteration
Selection of transformed embryonic stem cells
Cells taken up foreign DNA acquire
Antibody resistance
Select colonies of cells with identical genetic alterations
Isolation of transformed embryonic stem cells
Millions/billions of cells that can freeze down to liquid nitrogen
Expand transformed embryonic stem cells
Comprised of 2 completely different breeds
Not crossbred
Chimera
Sheep/goat chimera is a
Geep
Genetic alteration disadvantage
2 generations needed to get genetically engineered animal
Generic alteration advantage
Can target genetic alterations before the animal is made
Clone donor cells examples
Ear punch
Liver
Embryonic stem cells
1st cloned sheep using somatic cells
Dolly
What somatic cells were used to clone dolly
Mammary gland cells
Best candidate for transferring organs into humans
Pigs
1st cloned cat
Carbon copy
Cloning advantage
Can make a genetically engineered animal in one generation
Cloning disadvantage
Not super efficient
Expensive
UNL genetically engineered pigs contained what protein
Green fluorescent
Approaching cow for rectal palpation procedure
- Grasp tail firmly lift to one side
- Put on lubricated glove or sleeve
- Insert closed hand into rectum halfway to elbow
Heat checking: gilts exposed to boar exposure for __ to ____ minutes
15 to 20 minutes
The gilt being mounted by other gilts is
In heat
In heat gilt characteristics
Frozen, immobile, stands solid
Ears pinned back
Allows to sit on back
Red, swollen Vulva
How do you mark gilts in heat
Heat chemical symbol on back
Heat checking scoring system: dashed line
Didn’t show any signs of heat
Heat checking scoring system: 2++
Vulva is super swollen
Heat checking scoring system
-, 2-, 2, 2+, 2++
Triangle is what symbol
Chemical heat symbol
Gilts average ____ days of estrus cycles
21
Gilt AI tip is in what shape
Shape of boar penis
How do you insert an AI rod into a gilt
Upward motion, turn counter-clockwise
Estrous synchronization advantages
Decrease labor and calving period
Reduce calving season
More uniform weaning weights
Agonists for steroid hormone progesterone
Progestogens
Mimics luteal phase by having a light level of
Progesterone
Ways to administer progesterone
Injection
Feed
Implant
CIDR
CIDR application process
- Insert CIDR loading device into the vagina
- X-ray of CIDR loading device in vagina
- Release CIDR apparatus
- Leave a little bit of a tag to remove CIDR when necessary
Protocol of eazi-breed CIDR (beef cow)
Day 1: insert CIDR with CIDIROL capsule
Day 6: inject PG
Day 7: remove CIDR
Day 8: inject 2mL CIDIROL
Day 9-11: detect estrus
Synchro Mate B protocol (beef cattle)
Day 1: implant and inject SMB
Day 9: remove implant 48hr calf separation
Day 11-12: detect estrus
Danny is from
Eastern colorado
Danny has ___ pet cows
23
Danny’s birthday is
February 28th
Dr.white has ____ replacement heifers
900
MGA + lutalyse for synchronization protocol 1
Day 1: feed MGA
Day 14: MGA removed
Day 14-17: estrus, do not breed
Day 18: lutalyse injection, breed
MGA + lutalyse for synchronization protocol 2
Day 1: feed MGA
Day 8: inject lutalyse
Day 9: MGA is removed
Day 11-13: estrus detection, breed
Progesterone plateaus once a ____ ___ is formed
Mature CL
_____ occurs to start next estrus cycle
Luteolysis
Lutalyse protocol: 2-injection method
Day 1: inject lutalyse
Day 11-14: heat check and breed
Day 15: inject lutalyse
Lutalyse protocol: 1-method injection
Day 1: inject lutalyse
Day 11-14: heat check and breed
Lutalyse protocol: 1-injection + heat detection
Day 1-5: heat check
Day 6: inject lutalyse
After: heat check and breed
Older CL have a
Larger mass
Smaller CL respond
Quicker
Which reacts sooner: heifers or cows
Heifers
What can cause an abortion
PGF2aplha
What cattle respond poorly to the two injection system of PGF2alpha
Brahman
What is used mostly in problem breeding cows (dairy cows)
Cystorelin
Synchronizing of estrus and ovulation in cows steps
- PGF2alpha - cow ovulates
- PGF2alpha - new follicular phase
- GnRH
- PGF2alpha
- GnRH
- Insemination: 30-40% conception
Synchro-mate B protocol (sheep)
Day 1: insert norgestomet implant
Day 13: remove implant
Day 15: detect estrus
Easy-breed CIDR protocol (sheep)
Day 1: insert CIDR
Day 12: remove CIDR
Day 14: detect estrus
What animal can be effectively synchronized with P.G. 600
Prepubertal gilts
A.I. cow steps
- Make sure you can put a finger on cervix
- Put rod in vagina in upward motion and level
- Go through rings and expel semen in uterine horns
A.I. mare steps
- Use finger to feel opening of cervix
- Insert rod through cervix
- expel semen in uterine body
A.I. dog steps
- Insert AI rod fairly close to cervix
- Expel semen - enters uterine body
Bull semen evaluation calculation
100(50)N*1000
100 = dilution factor
50 = hemacytometer factor
N = # of sperm counted in hemacytometer
1000 = multiplication factor to represent 1 mL
Boar semen evaluation calculation
1050N*1000
Total number of sperm/ejaculate equation
Total # sperm/ejaculate = sperm concentration * total volume
Mechanism of action for stimulating the ejaculation process steps
- Intromission
- Sensory stimulation of glans penis
- Sudden and powerful contraction of urethralis, bulbospongiosus and ischiocavernosus muscles
- Expulsion of semen
Sensory stimulation of glans penis signals
Temperature
Pressure
Bull type of copulatory
Short
Bull ejaculation time
A few seconds
Bull volume of ejaculate
Fairly small
Bull concentration of sperm within ejaculate
Fairly high
Stallion type of copulator
Intermediate
Stallion mating time
30 seconds to 1 minute
Stallion volume of ejaculate
Much higher than bull
Stallion concentration of sperm
A little lower than bull
Boar type of copulator
Sustained
Boar mating time
5 minutes to 20 minutes
Boar volume of ejaculate
Significantly higher than bull and stallion
Boar sperm concentration
A lot lower than bull and stallion
3 muscles primarily associated with ejaculation
Ischiocavernosus
Bulbospongiosus
Urethralis
What are toxic to sperm
Powder and latex
What kind of glove is needed to collect semen from boars
Powder-free vinyl glove
Collecting semen from boars: use a
Phantom mount/dummy
Tease with female in estrus
Collecting semen from stallion
Artificial vagina integrated into a phantom mount
Phantom mount for a bull
Cowhide over it
Portable
Usually treated with hormones to mimic female in estrus and allow bull to mount
Jump steer
Head of sperm abnormalities
Crater defect
Tapered heads
Ruffled acrosome
Knobbed acrosome
What is the best fertility indicator
Motility
Tail abnormalities
Coiled tail
Double mid piece
Folded tail
Detached head
Sperm mature as they move through the
Epididymis
To tell if sperm is mature what is the key indicator
Cytoplasm droplet is as far as it moves from head of sperm then it is shed
Want to see less than __% of ejaculate consisting of abnormalities
20
Undifferentiated cells that replenish or repopulate either a specific tissue or organ or multiple tissues and organs
Stem cells
Hierarchy of stem cells
Totipotent
Pluripotent
Blood stem cells, muscle, nerve, bone, other tissues
The ability of a single cell to differentiate and develop into a complete organism
Totipotency
The ability of a single cell to differentiate into more than one cell type or tissue
Pluripotency
What opened doors for stem cell research
Dolly the sheep clone
Day 0-2 early embryonic development
Location: oviduct
Development: one cell
Day 1-3 early embryonic development
Location: oviduct
Development: 2 cell
Day 2-3 early embryonic development
Location: oviduct or uterus
Development: 4 cell
Day 3-5 early embryonic development
Location: uterus
Development: 8 cell
Day 4-5 early embryonic development
Location: uterus
Development: 16 cell
Day 5-8 early embryonic development
Location: uterus
Development: morula
Day 6-7 embryonic development
Compact morula
Day 7-8 embryonic development
Early blastocyst
Day 7-9 embryonic development
Blastocyst
Day 8-10 embryonic development
Expanded blastocyst
Day 9-11 embryonic development
Hatching blastocyst
Embryonic stem cells grow in
Colonies
Mono layer of cells that line the bottom of the culture plate
Fiberblast cells
Provide nutrients to the media that are required for the embryonic stem cells
Fiberblast cells
Efficiency of embryonic stem cell- like culture formation from porcine embryos
Less than 1% to develop embryonic stem cell
Form blood and immune cells
Responsible for constant renewal of blood
Isolated from blood or bone marrow
Hematopoietic stem cells
Hematopoietic stem cell characteristics
Can renew themselves
Can differentiate into a variety of cell types
Can move out of bone marrow into circulating blood
Rich in stem cells
Immunologically naive
Perfect match for donor
Life saving benefits for siblings
Differentiate into a variety of cell types
Umbilical cord stem cells
Umbilical cord stem cells are isolated from
Umbilical cord blood
AI gilts every __ hours while in heat
12
Injectable anesthetic cocktail
TARK
How is TARK administered into the ear vein
23 gauge butterfly cannula
What tubes are placed into the nares of the pig
Endonassal
What scrub is used to prepare surgical site
Betadine
Cleanse surgical site with
70% alcohol
What is removed from incubator and poured into syringe
BECM flushing media
What anesthesia is the pig maintained with
Isoflurane inhalation
What part of the scapal do you use
Curve part
__ inch skin incision is made on abdomen between _______ and last pair of _____
3
Umbilicus
Teats
Insertion of flushing cannula into _______ of the oviduct
Infundibulum
The cannula is held into a ___ mL ruble that will collect the flushing media and the embryos
50
____ -end needle administering media into the oviduct
Blunt
First __ mL will collect embryos
5
_____ appearance indicates rupture of CLs at ovulation and release of eggs
Bloody
Closing body wall use a ________ needle
Non-traumatic
Each suture is
Double knotted
Suturing skin stitch
Continuous
Suturing skin needle
Cutting edge
Body wall needle
Large curved
Indicator of pregnancy
Feel over pelvis to find cervix
Placenta of fetus feels like a bag of walnuts
Feel fetus
Day 21 embryo of calf
Heart begins to beat
Lungs, liver and pancreas begin to develop
Elongation of pig blastocytes shape at day 10
Spherical embryo
Elongation of pig blastocytes shape at day 11
Oval shaped embryo
Elongation of pig blastocytes shape day 11.5
Tubular shaped embryo
Elongation of pig blastocytes shape day 11.75
Elongated filamentous thread
Highest growth rate an individual will have their entire lifespan between day ___ and ___
10-12
Origin: Early entodermal layer
Yolk sac
Function: vestigial
Yolk sac
Origin: Cavitation From inner cell mass
Amnion
Function: Encloses fetus in a fluid-filled cavity
Amnion
Origin: Diverticulum of hindgut
Allantois
Function: Blood vessels connect fetal with placental circulation
Allantois
Origin: Trophoblastic capsule of blastocyst
Chorion
Function: Encloses embryo and other fetal membranes
Chorion
Origin: Amnion wraps about the yolk stalk
Umbilical cord
Function: Encloses Allantoic vessels
Acts as the vascular link between mother and fetus
Umbilical cord
Animals with a diffuse placenta
Pig
Horse
Lemur
Placenta where villi are spread evenly across the surface of the chorion
Velvety appearance
Diffuse
Animals with a cotyledonary placenta
Cow
Ewe
Deer
Placenta with villi grouped in rosettes and have circular structures on surface of chorion
Cotyledonary
Animals with a zonary placenta
Dog
Cat
Placenta where the villi are in a band-like appearance distribution
Zonary placenta
Animals with a discoid placenta
Rabbit
Rodents
Bats
Insectivores
Human
Placenta where one or two areas where villi are distributed and limited to disc-shaped areas
Discoid
Fetus becomes _____ when the water breaks
Hypoxic - starving for oxygen
Directly surrounds fetus
Inner-most layer
Amnion
Placentome consists of
Maternal caruncle
Fetal colyledon
Outermost water sac
Chorio-allantois
Innermost water sac
Amnio-allantois
Marker of length of placenta in pigs
Necrotic tip
When villi don’t shrink during birth it can cause
Retained placenta
What are used to help with retained placentas
Antibiotics and oxytocin
TARK meaning
Telezol
atropine
rompun
ketamine