Preservation of Bull Semen at Sub-Zero Temperatures — John Stuart Mill — John Shaqi
Preservation of Bull Semen at Sub-Zero Temperatures
John Stuart Mill · en
Temperatures in the immersion bath were recorded in a 2-milliliter glass
ampule containing 1 milliliter diluted semen and in an 8-milliliter
plastic vial containing 2.5 milliliters of diluted semen. A second
plastic vial and glass ampule filled to capacity with diluted semen
showed a cooling rate almost identical to that shown in Figure 4. It was
obvious from the comparison that samples in the plastic vials cooled
slower than those in glass and that the volume of semen (at least the
small volumes used) in the vials had little effect on the rate of
cooling. In another experiment, it was shown that the volume of diluted
semen in the ampule to be frozen (0.2, 1.0 or 5.0 ml.) had little or no
effect on the survival of the sperm.
STORAGE TEMPERATURE
In freezing and storing bull sperm, an alcohol bath containing dry ice
at a temperature of -79 deg. C. has been used as a cooling agent. In many
areas, the availability of dry ice is limited and the cost is rather
high. Mechanical means are available for obtaining temperatures as low
as, or lower than, -79 deg. C. but for the most part they are expensive. If
warmer temperatures were suitable for storing frozen semen, the ordinary
deep-freeze, which operates at -15 deg. to -25 deg. C., might be used.
=Storage at temperatures from -23 deg. to -79 deg. C.= In testing the effects of
storage temperatures on the survival of frozen bull sperm (in a diluent
containing 7 percent glycerol), 9 ejaculates were frozen and kept at
-23 deg., -37 deg., -51 deg., -65 deg., and -79 deg. C. The desired temperatures were
maintained by dropping pieces of dry ice into ethyl alcohol baths as
needed. Samples were thawed after 1 hour, 1 day, 3 days, and 5 days.
After 1 hour, the samples maintained at the various temperatures
exhibited approximately equal motility (Fig. 5).
[Illustration: Effect of freezing and storing bull sperm at various
temperatures on the sperm motility at thawing (average of 9 ejaculates)
(Fig. 5)]
At the end of 1 day, samples stored at -79 deg. C. exhibited approximately
the same motility as did similar samples stored for 1 hour. The samples
stored at -65 deg. C. had declined slightly in motility and those maintained
at -51 deg. C. had only one-third the motility which they had displayed at 1
hour. The samples at -23 deg. and -37 deg. C. exhibited practically no motility
after 1 day in storage. After 5 days, only 3 of the 8 ejaculates stored
at -51 deg. C. showed motility upon thawing. Apparently detrimental changes
take place more rapidly when the samples are stored at temperatures
warmer than -65 deg. C. The nature of these changes has not been determined.
Reports from other laboratories indicate that storage temperatures much
lower than -79 deg. C. are just as satisfactory as -79 deg. C.