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LECTURE I

ANS 416

ANIMAL BIOTECHNOLOGY

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Course Outline

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ANS 416: Animal Biotechnology

Definition of biotechnology; use of biotechnology in reproduction;

artificial insemination, embryo transfer; cloning and its concepts;

transgenic animals and their value; conservation of genetic

materials; biotechnology in animal nutrition: Types of rumenmicroflora, modification of rumen microflora through nutrition and

biotechnology; probiotics and enzymes; metabolic modifiers:

types (e.g. somatotropins, bovine placental lactogen, conjugated

linoleic acid) and effect on animal performance and quality of

food products (e.g. designer eggs); chelation and utilization of

inorganic elements; biotechnology and animal health; vaccines;

oligosaccharides and competitive inhibition; safety and ethical

issues in animal biotechnology; patenting issues.

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Definition of Biotechnology

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While biotechnology has been defined in many forms in essence it

implies the use of microbial, animal or plant cells or enzymes tosynthesise, break down or transform materials.

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Biotechnology is MULTIDISPLINARY industry

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USE OF TECHNOLOGY IN

ANIMAL PRODUCTION

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Somatic

cell culture

Nuclear transfer

(cloning)

Vaccines

Hybridomas

ProbesMonoclonals

diagnostics

Artificialinsemination

Probiotics

Enzymes / hormones

Growth

promotion

Unlimitedasexual multiplication

QuickTim e a nda TIFF(Uncompressed) decompressor areneededt oseethispicture.

Limited

asexual multiplication

Altered

properties

Micro-injection

Transgenic embryo

Re

combinant

DNA

Animal Biotechnology

In vitro

fertilisation

Embryo transfer

SuperovulationOocytes

Deep freezing

Embryo splittingIn vitro embryo culture

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Artificial insemination?

Artificial insemination is the placing of semen inthe female reproductive tract by

artificial techniques.

The use of this technique has

increased over the past several years

due to advancements in

technology.

The process varies by species, but it typically

involves specialized equipment and a trained

technician.

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Steps involved in artificial insemination?

The technician deposits a prepared unit of semen

into the reproductive tract of the female at a

predetermined time.

The objective of artificial insemination is to deposit

the semen at the appropriate time to achievefertilization.

Timing varies by species of animal and should be

closely studied prior to implementing artificialinsemination.

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There are several different segments that support

the process of artificial insemination.

Some of the major segments include:

A. Semen CollectionArtificial insemination

requires that semen be collected from the male.

Semen may be collected via manual stimulation or

theuse of an artificial vagina.

The technique used depends on the species from whichthe semen is being collected.

A. Semen Collect ion

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B. Semen evaluationAfter semen is

collected from the male, it

is evaluated for quality.

Only the highest quality

semen is utilized for

artificial insemination.

The semen must be evaluated under amicroscope to verify that it contains an

appropriate number of living sperm cells.

If approved for use, the sperm is then

processed to increase utilization.

B. Semen evaluat ion

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C. Semen extension

Semen is extended during theprocessing phase, which allows for a singlecollection to be divided into many units.

Only one unit is required for artificial insemination; byextending it, the semen can be used for severalbreedings.

To extend semen, specialized compounds are added to

enhance the viability and storage of the sperm cells. Depending on the species, often nutrients are added to

expand the mass of each semen unit.

C. Semen extension

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D. Semen storage

Semen is stored in one of

two ways.

It is either frozen to extremely low temperatures or

maintained fresh until use.

How semen is stored depends on the species.

For example, cattle semen is slowly cooled and frozen

at a temperature of320F (196C).

Liquid nitrogen is used to obtain these extremetemperatures. Properly frozen and stored semen canremain viable and be used 30 to 40 years later.

The frozen semen is thawed prior to use in a 95 to

98F (33 to 34C) water bath.

D. Semen sto rage

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E. Sexing semen

X-chromosome-bearing bull sperm (which lead to females) have

about 4% more DNA than Y-chromosome-bearing ones (whichlead to males). By placing sperm in a solution of DNA-binding dye,

X-chromosome-bearing sperm become more brightly stained than

Y-chromosome-bearing sperm. By use of lasers plus a device

called a cell sorter, it is possible to separate the sperm into threetest tubes, male, female, and unsexable (the great majority).

Those in the sexed test tubes accumulate at about 1,000 sperm

per second, with an accuracy of just over 80%.

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+ -+-

LASER

900

X sortY sort

Sperm

Stained

With DNA

SensitiveFluorescent

Dye

Flow Cytometer Separation of X and Y Sperm

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Advantages of AI

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Disadvantages of AI

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EMBRYO TRANSFER

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Embryonic stages

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Early stages of mammalian embryo developmen

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What is embryotransfer?

Embryo Transfer This procedure is when an

embryo in its early development is

transferred from its own mother to a recipient

mother.

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Embryo Transfer

Genetic progress is achieved fairly quickly than with

artificial insemination.

AI limits progress to male side of genetic

contribution

Embryo transfer allows each a cow to produce many

offspring and offers more rapid genetic gain.

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How is embryo transfer done?

1. The process of cattle embryo transfer begins by selecting

a genetically outstanding, well-nourished, non-pregnantembryo donor (heifer or cow). At the same time, 10 to15 non-pregnant females (embryo recipients) areidentified.

2. Super ovulation

The donor cow is super ovulated byusing injectable hormones. An average donor cowproduces 6 transferable embryos. However 0-25embryos can be expected.

3. Insemination

Cattle that are super ovulated are heatdetected and inseminated at both 12 and 24 hours afterheat with two straws of semen.

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4. After 7 days, the embryos have developed into atransferable state.

5. When the 7 days are up, a special solution is pushed intothe reproductive tract of the donor and the embryos arewashed out of her.

6. The solution that comes out of the tract is taken into alaboratory and searched for embryos.

7. The embryos can be freshly transferred into another cow

that is in the same part of her cycle as the donor or, theycan be frozen.

How is embryo transfer done (2)

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Artificial

insemination

Embryo transfer : common practice

Donorfemale

Recipientfemale

Superovulation(PMSG and/or FSH)

Induction of oestrus(PGF2a)

Selection

Manipulation

Cryopreservation

Embryo placementEmbryo collection

- surgical (sheep)

- non-surgical (cattle)

Synchronized

1 x 100 x

PMSG: PregnantMare Serum

Gonadotropin

FSH: Follicle stimulating hormonePGF: Prostaglandin F

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ET -

Scheme

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Stimulating Follicular Development

Estrus

Estrus

eCG or

FSH

ProgesteroneFrom C.L.

First FollicularWave

10-12

Multiple

Ovulations

PGF2a

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Foley catheter

Straw (0.25 ml)

Embryo collection

Cryopreservation

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What are the benefits of embryo transfer?

To produce more offspring from a valuable dam.

Some infertility problems can be avoided.

Prevents disease transmission.

Reproduce some endangered species faster thannormally possible.

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What are the limitations of embryo transfer?

Decreases genetic diversity

Decreased diversity often leads to mutations

Expensive and time consuming procedure

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Embryo Transfer

Who is this practical for?

Seed stock producers

An animal of genetic superiority whose offspring is demanded

by the industry.

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Course Outline

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ANS 416: Animal Biotechnology

Definition of biotechnology; use of biotechnology in

reproduction; artificial insemination, embryo transfer;

cloning and its concepts; transgenic animals and their value;

conservation of genetic materials; biotechnology in animal

nutrition: Types of rumen microflora, modification of rumenmicroflora through nutrition and biotechnology; probiotics and

enzymes; metabolic modifiers: types (e.g. somatotropins, bovine

placental lactogen, conjugated linoleic acid) and effect on animal

performance and quality of food products (e.g. designer eggs);chelation and utilization of inorganic elements; biotechnology and

animal health; vaccines; oligosaccharides and competitive

inhibition; safety and ethical issues in animal biotechnology;

patenting issues.