REPRODUCTION - Facultad de Ciencias Veterinarias

* Thermo Resistance Test
850
REPRODUCTION
surface; in the absence of freezing machines this step can be done in a large semen storage tank or a big polystyrene
container, containing liquid nitrogen); Place racks in liquid nitrogen at -196oC; Collect straws with a gloved hand and
store in goblets in liquid nitrogen; Wash and sterilize glassware for the next day. The common types of packaging used for
processed semen are: French Cassou straws packaged and sealed in straws, mini (0.25 ml) or medium (0.5 ml); German
minitub-packaged and sealed with small glass or steel spheres (0.25 ml). In both systems, each straw must contain a
minimum of 20–30 million spermatozoa per dose.In the Figure 1, it can be follow the different steps from the ejaculation
throughout the thawing process.
Diluters used for deep freezing semen
There are many diluters available for deep freezing buffalo semen. Some drawn from international literature; others
developed at regional level. Basically egg yolk and / or milk are the basic ingredients while glycerol is the main cryoprotective
agent used @ 7 per cent. Some vegetables derivate compounds, like coconut water and soybean milk have also
been used on experimental basis. The main diluters used in our work in Brazil and in other countries of Latin America have
its composition as follow:
Composition of TES (hydroxy-methyl-amino-ethan) diluter
Solution A - 500 mL (distillate or deionised water)
TES (hydroxy-methyl-amino-ethan)…………………………………………….24.5 g
TRIS (hydroxy-methyl-amino-methan).………………………………….. ...........5.3 g
D (-) Fructose …………………………………………………………………….1.08 g
Streptomycin …………………………………………………………………….700mg
Penicillin…………………………………………………………………………..70mg
Solution B - 100 mL (distillate or deionised water)
Skim milk…………………………………………………………………………..11g
Note: maximum heated to 80 ° C to not denature the protein contend
Preparation of the final diluter – 100 ml:
Solution A ……………………………………………………………………..36.5 mL
Solution B………………………………………………………………………36.5 mL
Glycerol………………………………………………………………………… 7.0 mL
Egg yolk……………………………………………………………………… 20.0 mL
Osmolarity to 290 ± 5mOsm
pH= 6.8 – 7.0
Proceedings 9 th World Buffalo Congress