Vie CONG. INTERN. REPROD. ANIM. INSEM. ARTIF., PARIS, 1968, VOL. I

frame into a water bath filled with 2.9 gram % Sodium citrate solution maintained at 35° C.

After 10 minutes the frame was removed from the water bath and placed in a similar
second bath. After another |0 minutes the frame was removed and the dialysing tubing was
carefully unwound from the frame and dried with a warm towel. The semen was carefully re-
moved by snipping one end of the tubing and used for insemination. Only 0.! cc of semen was
used per insemination.

RESULTS AND DISCUSSION

The results of the freeze recoveries are given in Table |. Semen, when collected direct~
ly into the extender and mixed immediately, showed very good free motility and did not coagu-
late. Fertility recorded was for all eggs laid by the hens from the 2nd day after insemina-
tion to 10th day. Semen frozen without glycerol gave zero to 6% fertility. Semen frozen
with glycerol and dialysed gave many fertilized eggs. Of the twenty eggs laid by three hens
only three hatched to give healthy chicks.

The semen that recovered after freeze-thawing though low in percent motility was healthy
and quite progressive. Because of the heavy freeze damage the semen that was inseminated had
an excess of dead sperm cells and this could have caused some deleterious effect on the fer-
tility potential of the normal cells. Turkey semen can withstand sudden temperature drops
(cold shock) after extension. Turkey semen can be frozen without glycerol or with quite low
concentration of glycerol and with little or no glycerol equilibration times. Vegetable pro-
tein source, like soy protein under the above circumstances, can afford the required cryo-
protective effect on the freeze-thaw processes.

REFERENCES
I. Wilcox, F. H. and C. S. Shaffner. Storage of Turkey semen, Poultry Sci. 39:1580 (1960).

2. Harris, G. C., Hobbs, T. D., Brown, J. E., and L. B. Warren. The storage of turkey
spermatozoa in Sodium citrate and Carbondioxide extenders. Poultry Sci. 42:536. (1963).

3. Bajpai, P.l. and K. |. Brown. The effect of egg yolk on semen characteristics of turkeys.
Poultry Sci. 42:888. (1963)

4, Shaffner, C. S. Observations on freezing chicken semen. Vth International Congress of
Animal Reprod. 3:426 (1964).

5. Graham, E. F. Personal Communication (1965).

6. Polge, C. Functional survival of fowl spermatozoa after freezing to -79°C.
Nature, Lond. 167:949 (1951).

7. Rajamannan, A. H. J. and E. F. Graham. Spin freezing of semen. VIth Inter. Congress
of Animal Reprod. (1968).

8. Rajamannan, A. H. J. and E. F. Graham. Film freezing of semen. VIth Inter. Congress
of Animal Reprod. (1968).

9. Rajamannan, A. H. J. A new freezing technique. A. |. Digest I4:18 (1966).

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