Lab Final Flashcards by Miranda Unknown

Reproductive technologies

Semen sexing
IVP of embryos
Transgenic animal production
Gene mapping
Marker assisted selection

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Transgenic animal production examples

Micro injection
Blastocyst injection
Nuclear transfer

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Propagate superior livestock animals

Cloning

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Remove or add a gene to improve a breed of livestock

Transgenics

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Select animals for specific traits very early in life

Coat color in cattle

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Production of proteins which are essential to human life

Milk
Blood
Urine

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Alter specific genes in specific locations to attain a desirable result

Transgenics

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What chromosome gives off more light emission because it has more dye and more DNA

X chromosome

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What chromosome has an extra short arm

X chromosome

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What determines sex

Sperm

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What excites vital dye in semen sexing

Laser

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Why does the X chromosome have more dna

Extra short arm

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Sperm run through system ______

Individually

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Sperm is given a charge based on

Light intensity

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Pass magnetic field in semen sexing

Sorting

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Dead or damaged sperm not selected

Purifying samples

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System that has a 95% or better success sorting

Flow cytometric analysis of sperm

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Ovaries isolate

Oocytes

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Step 1 of in vitro fertilization

Synchronization of recipients with donor

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Step 2 of in vitro fertilization

Superovulation of donor

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Multiple treatments of porcine FSH promotes more

Follicle development

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8 CLs mean the donor is

Super ovulated

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Step 3 or in vitro fertilization

Inseminate donor with semen from genetically superior bull

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Work AI rod through the ______ by ______ ________

Cervix
Rectal palpation

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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

1. Grasp tail firmly lift to one side 2. Put on lubricated glove or sleeve 3. 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

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

1. Insert CIDR loading device into the vagina 2. X-ray of CIDR loading device in vagina 3. Release CIDR apparatus 4. 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

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

1. PGF2alpha - cow ovulates 2. PGF2alpha - new follicular phase 3. GnRH 4. PGF2alpha 5. GnRH 6. 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

1. Make sure you can put a finger on cervix 2. Put rod in vagina in upward motion and level 3. Go through rings and expel semen in uterine horns

A.I. mare steps

1. Use finger to feel opening of cervix 2. Insert rod through cervix 3. expel semen in uterine body

A.I. dog steps

1. Insert AI rod fairly close to cervix 2. 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

10*50*N*1000

Total number of sperm/ejaculate equation

Total # sperm/ejaculate = sperm concentration * total volume

Mechanism of action for stimulating the ejaculation process steps

1. Intromission 2. Sensory stimulation of glans penis 3. Sudden and powerful contraction of urethralis, bulbospongiosus and ischiocavernosus muscles 4. 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

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

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

____ -end needle administering media into the oviduct

Blunt

First __ mL will collect embryos

_____ 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