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Counting dead spermatozoa in frozen semen

Published online by Cambridge University Press:  02 September 2010

H. R. L. Buttle ,
J. L. Hancock  and
A. F. Purser
H. R. L. Buttle
Affiliation:
A.R.C. Animal Breeding Research Organisation, Edinburgh 9
J. L. Hancock
Affiliation:
A.R.C. Animal Breeding Research Organisation, Edinburgh 9
A. F. Purser
Affiliation:
A.R.C. Animal Breeding Research Organisation, Edinburgh 9
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Summary

Three methods were used to make differential counts of living and dead bull spermatozoa in samples of frozen semen in an egg-yolk citrate medium containing glycerol.

With two methods (‘phase’ and ‘nigrosin’ methods) dead spermatozoa were identified by their altered structure. With a third method (nigrosineosin) dead spermatozoa were identified by their staining affinity. With the phase method spermatozoa were immobilised by treatment with approximately M/40 sodium fluoride and were examined by phase-contrast microscopy in unfixed wet preparations. With the other two methods the spermatozoa were examined in smears stained either with nigrosin alone (nigrosin method) or with nigrosin and eosin (nigrosin-eosin method).

Analysis of variance of the results of a factorial experiment involving fluoride-treated and untreated samples from 6 bulls with the three methods showed that differences between semen samples contributed 66% of the variation. A defect of the phase method was that the variance between counts was greater than the theoretically expected value.

All spermatozoa in nigrosin-eosin stained preparations were stained with eosin within a few days of the smears being made, so that living and dead spermatozoa could not be distinguished by their differing affinities for eosin. Repeat counts on nigrosin-stained preparations did not differ significantly from counts made several days previously. Sodium fluoride in the concentration used here (M/40) tended to reduce the percentage of dead spermatozoa.

Type
Research Article
Information
Animal Production , Volume 7 , Issue 1 , January 1965 , pp. 59 - 65
Copyright
Copyright © British Society of Animal Science 1965

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References

REFERENCES

Beatty, R. A., 1957. Nigrosin-eosin staining of rabbit spermatozoa and the fertility of semen. Proc. roy. Soc. Edin. (B), 67: 1.Google Scholar
Beatty, R. A., & Napier, R. A. N., 1960. Genetics of gametes. I. A quantitative analysis of five characteristics of rabbit spermatozoa. Proc. roy. Soc. Edin. (B), 68: 1.Google Scholar
Blackshaw, A. W., 1958. The effects of glycerol on the supra-vital staining of spermatozoa. Aust. Vet. J., 34: 77.CrossRefGoogle Scholar
Campbell, R. C., Dott, H. M., & Glover, T. D., 1956. Nigrosin-eosin as a stain for differentiating live and dead spermatozoa. J. agric. Sci., 48: 1.CrossRefGoogle Scholar
Campbell, R. C., Hancock, J. L., & Rothschild, Lord, 1953. Counting live and dead bull spermatozoa. J. exp. Biol., 30: 44.CrossRefGoogle Scholar
Campbell, R. C., Hancock, J. L., & Shaw, I. G., 1960. Cytological characteristics and fertilising capacity of bull spermatozoa. J. agric. Sci., 55: 1.CrossRefGoogle Scholar
Fisher, R. H., & Yates, F., 1949. Statistical Tables for Biological, Agricultural and Medical Research. Oliver & Boyd, Edinburgh.Google Scholar
Hancock, J. L., 1952. The morphology of bull spermatozoa. J. exp. Biol., 29: 445.CrossRefGoogle Scholar
Hancock, J. L., 1957. The morphology of boar spermatozoa. J. roy. Micr. Soc, 76: 77.CrossRefGoogle ScholarPubMed
Mixner, J. P., & Saroff, J., 1954. Interference of glycerol with differential staining of bull spermatozoa as used with semen thawed from the frozen state. J. Dairy Sci., 37: 1094.CrossRefGoogle Scholar