College

College of of
veterinary science
veterinary science
and
and animal
animal husbandry
husbandry

topic- storage and transport of semen

Presented By
Name – arti kumari

Roll number – 46/19

Batch – 2019
 Expected outcome

1. Short term storage of semen and storage of Frozen

semen
2. Handling during and after thawing of frozen
semen.
3. Transport of chilled semen and frozen semen
4. Liquid shipper vs vapour shipper
 SHORT-TERM LIQUID STORAGE OF SEMEN

Fertile spermatozoa can be preserved for a short period of time without freezing.

 The spermatozoa of many species can be stored in liquid media which is

maintained either at ambient temperature or in a refrigerator at 4-5˚c.
 There are definite advantages to store spermatozoa at ambient temperature,
especially where refrigeration facilities may not be readily available.
 For successful short term liquid storage of semen it is necessary neither to extend
the time that spermatozoa are able to maintain high metabolic rates or to slow
the process down.
 The first effective ambient temperature storage medium was based on the fact
that carbon dioxide markedly reduced the rate of sperm metabolism.
 This storage, medium which was termed Ilini Variable Temperature (IVT)
extender,was saturated with carbon dioxide by bubbling the pure gar through
an egg yolk –citrate extender.
Name of Diluter use at room temperature For preservation of semen: -

1. Coconut milk extender

2. Cornel University Extender
3. Illini variable temperature
 In this extenders semen sample of bull survive 2-3 days but in case of buck semen
survive only for 12-24hrs.

Name of Diluter/extender use for preservation of semen at refrigerator

temperature(4-6°c)

 Egg yolk phosphate

 Egg yolk citrate
 Egg yolk tris
 D2 diluter
 Milk Diluter
 Egg yolk glucose bicarbonate
 Citric acid whey(caw):-developed at NDRI to preserve the buffalo bull semen
- At refrigerator temperature semen can be preserve for 2-4 days.

Dilutor use at subzero temperature

 Glycerol egg yolk tris

 Glycerol egg yolk citrate
 Glycerol milk Dilutor
 Storage of Frozen Semen

 The aim of storage of semen is to prolong the fertilizing

capacity of spermatozoa by reducing or arresting their
motility and metabolic activity.
 Frozen semen is stored in liquid nitrogen at-196°C in LN2 container.
 The different sizes of liquid nitrogen containers are available in the
market.
 These range from 1 litre capacity of nitrogen to 550 litres or more.
 At the semen freezing lab large cryogenic containers are used for
storing of semen straw in bulk undisturbed for longer period.
 Smaller containers are used to store semen from which straws have to
be utilized frequently.
 STORAGE OF FROZEN SEMEN

 Artificial insemination workers use small capacity of liquid

nitrogen container ranging from 1 litre to 20 litre capacity
tank.
 The recommended minimum volume of Liquid Nitrogen
present in the tank is at least 5 cm (2 inches) in order to
prevent any damage to the stored specimens.
 Ideally, the Liquid nitrogen level should be monitored
every week with a calibrated measuring stick.
 The straws are first placed in pre-cooled goblet.
 The goblet is kept in the canister.
 The canisters are hanged inside the liquid nitrogen
container with the help of their long handles.
 Handling of semen in LN2 container

 In the upper half of the neck tube of LN2 container, high

temperatures exist.
 The typical temperature range in the neck of the LN2,
container is -191°C to +2.2°C Temperatures can reach
+54° F (12.2 °C) in the neck of the container (1 inch from
the top).
 If the entire canister of semen is withdrawn above the
frost line (3 to 4 inches from the top of the tank) All straws
of semen will be damaged.
 Thermal injury to sperm is permanent and cannot be
corrected by returning semen to liquid nitrogen.
 Remove the canister from its storage position to the middle of
the tank.
 Raise the canister just high enough in the neck region to grasp
the desired straw.
 Grasp the straw and immediately lower the canister to the
tank floor use tweezers or long pre cooled forceps to remove
the straw.
 The straw should be removed within 10 seconds from the time
the canister is raised into position.
 Then immerse the straw in 37°C water immediately, after the
straw is immersed in water.
 The canister should be returned to it storage position.
 Any time if it takes more than 15 seconds to take out the
desired number of straws. The canister should be lowered back
into the tank to cool completely and again after sometime
take out the canister.
 TRANSPORT OF chilled SEMEN

 Use sterile plastic tubes for the semen which do not break during the transport.
 The chilled semen is usually sent in an ordinary thermos flask, or a styrofoam box
with ice-packs.
 It is imperative that the temperature does not drop below 0°C to ensure that the
semen does not freeze.
 Therefore, the tube containing the semen must be protected from direct
contact with ice cubes or cold-packs, for this, the tube is wrapped in a piece of
cotton wool.
 TRANSPORT OF frozen SEMEN
The two principal refrigerants used in experimental and routine shipping of frozen semen in recent
years dry ice (Co2) and liquid nitrogen.

Dry ice functions at -110˚ F, (-79˚C.) and liquid nitrogen at -320˚F, (-196˚C), and hence the latter has a
longer safety factor.

The dry-shipper is specially designed for shipment of frozen semen.

The various sizes of the dry shipper are commercially available

The shipper absorbs the liquid nitrogen into a porous material in its walls

These will not spill and therefore, need not be shipped as dangerous goods, which is more expensive.

They should, however, always be sent as fragile goods because they are easily broken by rough handling.

The tank is usually shipped in a plastic box for protection.

 TRANSPORT OF FROZEN SEMEN

 Care should be taken while transferring frozen semen

from storage to shipping containers.
 Exposure of frozen semen to room temperature should
not be more than a few seconds.
 This should be performed with shipping and storage
containers placed side by side.
 Always work within the neck of the storage container,
below the frost line, while locating, identifying, and
Grasping straws
 Liquid shipper vs vapour shipper

 If liquid shippers (liquid nitrogen containers) are tilted during transport, it will lose all liquid nitrogen
and warm rapidly, destroying valuable semen.
 Also, liquid nitrogen container is a safety hazard and could cause serious injury to handlers.
 Because of this, liquid shippers must be classified as containing hazardous materials, which requires
special paperwork and increases shipping costs.
 Vapour shippers, on the other hand, contain no "liquid" nitrogen.
 The liquid nitrogen vapour absorbed into the containers. Hence, vapour shippers are often referred
to as "dry shippers."
 No liquid nitrogen means no hazardous
materials classification and fewer shipping
restrictions.
 Vapour shippers are generally lighter than liquid
storage containers and are therefore also less
expensive to ship.
 MVE manufactures a "mushroom" shaped
protective shipping carton for all sizes of
shipping tanks.
 The carton has wide base with a rounded top
that reduces the likelihood of the tank tipping
over or being loaded upside down or on its side.
 The hard moulded plastic carton also helps
prevent tank damage due to normal shipping
use.