Artificial insemination

techniques in dairy cattle

Clinical conference II

Yana Mani Nepal

B.V.Sc & A.H
Roll No. 31
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 Artificial insemination
 very simply means taking sperm from the
best male animal and putting into the best
female animal.

used extensively in modern farming to

produce best diary and beef cattle and in
the breeding of race horses.
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It was in 1780 that the first scientific
research in AI of domestic animals, was
carried out on dogs.
Lazanno Spalbanzani, an Italian scientist,
conducted experiments that proved the
power of fertilization vested with the
spermatozoa and not with the liquid
portion of the semen.

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AI is a remarkable method of breeding
quality cattle in the most natural way
possible.

AI is being carried out in a large number of

buffaloes and cows and is extremely useful
in countries like Nepal, wherein quality
sires have been scarce.
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Interesting fact….
Semen volume/ejaculate: 6 ml

Sperm concentration: 1.2 billion/ml (motility: 70%)

Total sperm/ejaculate: 6x1.2billion ¡Ö 7.2 billion

Sperm motility of diluted, frozen and thawed semen: 40

Total post-thaw motile sperm/ejaculate: 0.40x7.2 billion= 2.88 billion

Post-thaw motile sperm required/insemination: 10 million

Breeding units/bull/day: 2.88 billion/10 million = 288 units

Intervals of semen collection: 3 days (95x) and 2 days (40x)

Semen collections/year: 95 + 40 = 135 times

Breeding units/bull/year: 135 times x 288 units = 38,880 ¡Ö 40,000

Reproductive life after progeny test: From 4 to 10 years old = 7 yrs.

Breeding units available from a tested bull: 40,000x7 ¡Ö 300,000 6

 Advantages of AI
 Use of proven stock quality
 Frozen semen can be transported globally and
stored for many years
 Cost effective - A bull is expensive to rear,
relatively unproductive, vulnerable to disease or
accident and may be infertile.
 Flexibility: high yield dairy cows can be crossed
with dairy bull semen to create more higher yield
cows.
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 Advantages contd…
• Safety: bulls are aggressive

• Disease control: Vibriosis, trichomonasis,

campylobacteriosis and brucellosis

•Use bulls with certain characteristics which

complement those of the particular cow (as
milk yield, milk fat%, or udder conformation
etc)

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Disadvantages
Main disadvantage is that the same alleles
keep appearing in the population so there
is a loss of diversity in the gene pool
(many genes are lost). So a disease could
wipe out an entire population if resistant
alleles have been lost.
Much time is required in checking females
for estrus
Some special facilities for corralling and
inseminating
Trained personnel  
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ARTIFICIAL INSEMINATION TECHNIQUES

1. Semen collection

2. Semen evaluation
3. Semen Processing, Storage,
Thawing, and Handling

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Semen collection

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Method of semen collection

1. Taking semen from vagina with spoon

or rubber breeding bag
2. Massaging vesicular gland and ampullae
by way of rectum
3. Artificial vagina
4. Electroejaculation

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Collection through artificial vagina

AV should be held parallel to the expected

path of bull's penis, after false mounts

Guide penis to the side by grasping sheath

and direct it into AV

Bull will thrust and ejaculate simultaneously

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FALSE MOUNTING: Deviating penis to side during mount. Sexually
stimulates the bull.
Providing two false mounts with two minutes of active restraint and one
additional false mount maximizes sperm cell numbers. 15
Electroejaculation
Principle: Electrically stimulating
nerves to male reproductive system
Uses for bulls
a. For crippled, aged or sexually low active dairy
bulls
b. For checking semen quality of beef bulls prior
to natural breeding season

Equipment: A bipolar electrode and a variable

source of alternating current: voltage: 0 - 30 V,
amperage: 0.5 - 1.0 A; alternating + and - rings
spaced 4cm apart, running longitudinally along
the electrode
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Electroejaculation procedure in bull
Restrain the bull in a chute with good footing provided

Remove excess faecal material from rectum

Insert lubricated electrode into rectum

Stimulate electrically for 2 to 5 min: low voltage at

gradual increase (a few volts at a time) up to 15 V
or higher, with 4-second rest period between
stimulations 17
Semen Evaluation
Evaluate semen quality by
volume,
colour,
consistency,
mass motility (overall movement
observed in the microscopy, "waves"),
individual motility of sperm cells

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Normal Parameters

Parameter Normal Values

Ejaculate volume 5 ml (range 1-15 ml)

Sperm concentration 1200 million/ml

(300-2500 million/ml)

Total sperm per ejaculate Typically 4-5 billion

Progressive motility Greater than 30%

Morphology Greater than 70% normal

http://www.vivo.colostate.edu/hbooks/pathphys/reprod/semeneval/bull.html
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Colour
 Opacity: Indication of concentration
 Colour: acceptable colour
ranges from milky to
creamy
 (Note: This indicates
sperm per cubic
millimetre of 500,000 or
above.
 Other colours indicating
less than 500,000 sperm/cu mm
would be opalescent (cloudy) to watery.
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Bulls: > 30% progressively motile sperm
Adversely affected by
heat
cold
residue on collection equipment
wrong pH or osmolality
sexual inactivity

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Semen Processing, Storage, Thawing,
and Handling
Processing steps of frozen bull semen
Dilution

Cooling

glycerolation & equilibration

packaging

freezing

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Semen dilution

Predilution
 Warm semen with 3 to 4 volumes of diluter
tempered in the same water bath
 Lecithin and lipoprotein from yolk of diluter:
Protect sperm from cold shock and prevent
changes in cell wall permeability during cooling
process

Dilution
After cooled to 50C , the pre-diluted semen is
diluted to final volume with diluter cooled to 50C

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Cooling semen
 Optimum cooling rate: From body temperature to 5oC for
1.25 to 2 hours (fast cooling) or for 2 to 4 hours (slow
cooling)

Glycerolation
 Sperm protection from freezing by glycerolation: Because
glycerol binds water, glycerolation results in

a. Decreased freezing point of solutions

b. Less ice formed
c. Correspondingly decreased concentration of solutes
d. Partially dehydrated sperm cells
e. Reduced selective freezing of water
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Semen packaging
Plastic straws
a. Size: 0.5 ml with 113 mm long and 2.8
mm in diameter (In Europe 0.25 or 0.3 ml
straws are used)

Freezing
Freezing procedure: A single layer of
straws are placed on a tray at 5.5 cm
above the liquid nitrogen level; Straws will
reach the temperature of liquid nitrogen
vapour in about 2 min.
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Storage of bull semen

Optimum temperature for semen storage

a. Liquid semen: at 50C
b. Frozen semen: below -750C

Semen storage in liquid nitrogen container

a. Double wall stainless steel or aluminium
container with a vacuum between the wall
b. Field unit
Liquid nitrogen capacity: 20 litres
Holding time: 90 days
Storage capacity: 1,200 straws of 0.5 ml capacity
Recharging interval: 60 days

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References
http://nongae.gsnu.ac.kr/~cspark/teaching/lecture_under.html
http://www.fao.org/AG/aga/agap/frg/FEEDback/War/u6600b/u6600b0m.htm#To
pOfPage
www.das.psu.edu/reproduction/insem
www.msue.msu.edu/msue/imp/modaa/16360001.html
http://www.buzzle.com/
www.vet.ksu.edu/studentorgs/bovine/pdf/artificial_insemination.pdf

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Thank You

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