Care and Handling of LN2 Containers

1. Containers should not be dragged on the ground, should

be lifted by handles and be placed gently on the ground.
2. They should not be welded.
3. Dont plug it tight while holding liquid gases.
4. Dont keep the containers dry and warm, frequent
cooling and drying reduces the durability.
5. Charge a warm container slowly and steadily.
6. After opening the containers, close it as quickly as
possible to prevent evaporation and wastage.
7. Clean them once in a year.
8. To read the level of liquid gases never use a hollow tube.
Use wooden stick with suitable markings and dip slowly to
the bottom, lift it out and wave it in the air and measure
the length of frost to read the level of the LN2 container.
9. Replenish as the level touches the lower one third.
10. Dont expose your face directly on the vapours.
Inhalation of Liquid Nitrogen vapour causes conjunctivities
and sometimes fainting.

11. Avoid violent handling; uneven flow and splashing will,

cause cold burn (Frost bite). Apply cold water in the event
of splashing on the body.
12. Canisters should be inserted into the refrigeration
slowly and carefully.
13. Should not handle the refrigerator roughly and avoid
tilting and drifting.

Handling of Frozen Semen Straws

1. Remove the neck plug of the container gently.
2. Cool the tip of the forceps sufficiently.
3. Lift the canister properly without disturbing other
canisters.
4. Dont lift the canister containing straws more than the
bottom level of the neck of the container.
5. Take one straw at a time with the cooled forceps within 5
sec.
6. Shake the straw, once or twice after holding with your
fingers to remove the drops of LN2 entrapped in the sealing
to avoid bursting.
7. Replace the canister and neck plug back in position.

How to do thawing

1. Thawing should be done quickly to avoid the death of the

sperms.
2. Straw should be immersed completely in the water with
the lab seal on the top.
3. Flask should be of sufficient height (15 cms). Every day
fresh water should be used.
4. A uniform thawing time and temperature of 30 seconds
at 37C should be followed.
5. After taking the straw it should be wiped free of water
using clean towel or absorbent cotton.

Loading of A.I Gun

1. Hold the straw by lab plug end and give one or two
shakes to bring the air space in the lab. plug end to prevent
wastage and contamination of semen while cutting that
end.
2. Put the factory plug end (double seal) down into the
barrel of the gun and load it.
3. With a pair of scissors cut the straw at the lab. seal end
over the air space at right angles.

4. If it is cut oblique, semen may leak into the sheath

during insemination. Avoid contamination of sheath at
every stage.
5. Slip the sheath over the gun and fix the sheath with O
ring.

Selection of Animal

Before taking the straw from the LN2 container, confirm the
animal for proper heat (External os opening, tone of uterus)
during rectal examination and dont handle ovaries.

Method of Inseminations

1.Hold the AI gun with 3 fingers.

2. Evert the vulval lips.
3. Introduce the AI gun at 45C and touch the floor of the
vagina.
4. Guide the tip of the AI gun to the external os and fix it.
5. Without applying force, slip the cervix on the gun.
6. Correct site of insemination using frozen semen is 50%
at the internal os and 50% in the mid cervix in the fresh
(first) insemination.
7. If repeated within 30days, the entire quantity is
deposited in the mid cervix itself.
8. Safe level for the frozen semen straw in the LN2
container is 15cms.
9. If it reaches 15cms never open the neck plug.
10. Arrange for topping up of LN2.
11. If any sweating is noticed on the external wall it
indicate that the container is defective.

GENERAL

Testing quality of the straw should be done only at the Lab.

1. Inseminate the cow as quick as possible after preparing

the AI gun.
2. Handle the cow and AI gun gently.
3. Dont expose the AI gun to sunlight.
4. Warm the AI gun by rubbing by hand during cold
weather.
5. Clean the vulval lips and evert it before inserting AI gun.
6. Inseminate at body of uterus and dont go into the
uterine horn.
7. Forward massage cervix gently, tickle clitoris and
massage well.
8. Train your attendant properly about correct handling of
frozen semen.