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Effect of freezing on bovine semen diluter components
ABSTRACTS,
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or without PGE, stored at +22”C for 24 hr. PGEl improved the life span of frozen PC. The addition of 100 ng of PGE1/ml of whole blood produced an improvement in the life span of either liquid-stored PC or previously frozen PC. Sumida, S. Preservation of human platelets for transfusion at +22” and -196”C, and prostaglandin E1. Low Temp. Med. 2, 131-135 (1976). Dayian, G., and Rowe, A. W. Cryopreservation of human platelets for transfusion. Cryobiology 13, l-8 ( 1976). Valeri, C. R., et al. Prostaglandin in the preparation of blood components. Science 175, 539542 ( 1972). SESSION
S7. pH
F. SPERMATOZOA, AND EMBRYOS
OVA,
Change of Bovine Semen Bugler with Temperature. R. S. JEYENDRAN AND E. F. GRAHAM (University of Minnesota, St. Paul, Minnesota 55108).
The pH change due to temperature was measured for a combination of Good’s buffers, Tes and Tris (TEST), with and without egg yolk containing 2 or 6% of 19 different cryoprotective compounds. The pH at 5°C was significantly higher than at room temperature. The addition of egg yolk and/or cryoprotective compound did not alter the pH significantly during cooling, even though a slight drop in pH was noted with the addition of 20% egg yolk, indicating that the change in pH is primarily due to the buffer. The pH change of 10 different buffering systems with temperature from 22 to 5°C was examined. Of these, three were conventional buffers, which included phosphate yolk, citrate yolk, and skim milk, and seven were Goods buffers with egg yolk, which included Tes, Tris, Bes, Mops, Pipes, Mes, and TEST. The results showed that the pH of the three conventional buffers did not change with decreasing temperature, but Good’s buffers showed an increase in pH with decreasing temperature from 22 to 5°C even though they had an excellent buffering capacity. (pK, of Good’s buffers changed very slightly with temperature. ) The pH of TEST yolk buffer (pH 7.2 at room temperature) was measured con’tinuously from 37°C to below freezing (-19°C). The pH increased linearly with decreasing temperature to pH 8.0 C 0.2 from 37°C to about -14°C at which point it dropped abruptly to pH 6.5 -t 0.2. The change in pH observed with decreasing temperature did not alter the viability of bovine spermatozoa during freezing and rewarming.
ANNUAL
IrlEETING
701
58.1 Effect of Freezing on Bovine Semen Dduter Components. CHRISTOPHER P. D'ALLEINXE* ASD C. P. MERILAN (University of hfissonri, Columbia, Missouri 65201). 59. Relation between Different Assays for Sperm Quality and Fertility of Frozen Boar Semen. &I. K. PAVELKO,* S. EIXARSSON," ASD B. G. CRABO (Department of Animal Science, University of Minnesota, St. Paul, Minnesota 55108). Fertility of boar semen appears to be poorly related to currently used laboratory techniques. This study attempted to correlate the fertility of frozen boar semen with an in vitro test measuring the uptake of ‘=I-labelled boar seminal proteins added to the thawing fluid. Semen from eight boars was frozen according to the Beltsville concentrated technique and thawed in OLEP (Larsson and Einarsson, Acta Vet. Scud. 17, 43, 1976), BTS (Purse1 and Johnson, J. Anim. Sci. 40, 99, 1975), and TESNaK. The first ‘two fluids have resulted in good fertility and TESNaK in zero fertility (Crabo, Larson, and Graham, Cryobiology 9, 331, 1972). Sperm thawed in OLEP featured the highest uptake of radiolabelled proteins (874%) of the plasma activity and those in TESNaK the lowest (402%). There was no difference in the percentage of normal acrosome (21-25s) or osmotically reactive cells (21-25s) between the fluids but the sperm motility was lower in TESNaK (8 vs 26% ). Furthermore, the fertility of the eight boars was assessed after insemination of 16-37 sows per boar with semen frozen as described above and thawed in BTS. The fertility varied from 23 to 71% between boars. The correlation between fertility of the individual boars and sperm motility was 0.19, between fertility and percentage normal acrosome ridges 0.29, and between fertility and the “protein uptake” in BTS 0.43. However, a correlation coefficient of 0.77 was obtained between fertility and the protein uptake in TESNaK. Thus, it appears that the less beneficial thawing fluid aided in disclosing differences between boars. It is possible that the composition of the thawing fluids affects the binding mechanisms of surface coating macromolecules in the sperm membrane. Alterations of the sperm surface may result in rapid elimination of the spermatozoa from the female tract. Differences between boars in this respect may account for loss of fertility by certain boars after freezing of the semen. 60. Acrosornal Proteolytic Actioity of FrozenThawed Boar Spermutozoa. K. I. LARS~~N * (Span. B. G. Crabo) (Department of Ob-
14TH
or without PGE, stored at +22”C for 24 hr. PGEl improved the life span of frozen PC. The addition of 100 ng of PGE1/ml of whole blood produced an improvement in the life span of either liquid-stored PC or previously frozen PC. Sumida, S. Preservation of human platelets for transfusion at +22” and -196”C, and prostaglandin E1. Low Temp. Med. 2, 131-135 (1976). Dayian, G., and Rowe, A. W. Cryopreservation of human platelets for transfusion. Cryobiology 13, l-8 ( 1976). Valeri, C. R., et al. Prostaglandin in the preparation of blood components. Science 175, 539542 ( 1972). SESSION
S7. pH
F. SPERMATOZOA, AND EMBRYOS
OVA,
Change of Bovine Semen Bugler with Temperature. R. S. JEYENDRAN AND E. F. GRAHAM (University of Minnesota, St. Paul, Minnesota 55108).
The pH change due to temperature was measured for a combination of Good’s buffers, Tes and Tris (TEST), with and without egg yolk containing 2 or 6% of 19 different cryoprotective compounds. The pH at 5°C was significantly higher than at room temperature. The addition of egg yolk and/or cryoprotective compound did not alter the pH significantly during cooling, even though a slight drop in pH was noted with the addition of 20% egg yolk, indicating that the change in pH is primarily due to the buffer. The pH change of 10 different buffering systems with temperature from 22 to 5°C was examined. Of these, three were conventional buffers, which included phosphate yolk, citrate yolk, and skim milk, and seven were Goods buffers with egg yolk, which included Tes, Tris, Bes, Mops, Pipes, Mes, and TEST. The results showed that the pH of the three conventional buffers did not change with decreasing temperature, but Good’s buffers showed an increase in pH with decreasing temperature from 22 to 5°C even though they had an excellent buffering capacity. (pK, of Good’s buffers changed very slightly with temperature. ) The pH of TEST yolk buffer (pH 7.2 at room temperature) was measured con’tinuously from 37°C to below freezing (-19°C). The pH increased linearly with decreasing temperature to pH 8.0 C 0.2 from 37°C to about -14°C at which point it dropped abruptly to pH 6.5 -t 0.2. The change in pH observed with decreasing temperature did not alter the viability of bovine spermatozoa during freezing and rewarming.
ANNUAL
IrlEETING
701
58.1 Effect of Freezing on Bovine Semen Dduter Components. CHRISTOPHER P. D'ALLEINXE* ASD C. P. MERILAN (University of hfissonri, Columbia, Missouri 65201). 59. Relation between Different Assays for Sperm Quality and Fertility of Frozen Boar Semen. &I. K. PAVELKO,* S. EIXARSSON," ASD B. G. CRABO (Department of Animal Science, University of Minnesota, St. Paul, Minnesota 55108). Fertility of boar semen appears to be poorly related to currently used laboratory techniques. This study attempted to correlate the fertility of frozen boar semen with an in vitro test measuring the uptake of ‘=I-labelled boar seminal proteins added to the thawing fluid. Semen from eight boars was frozen according to the Beltsville concentrated technique and thawed in OLEP (Larsson and Einarsson, Acta Vet. Scud. 17, 43, 1976), BTS (Purse1 and Johnson, J. Anim. Sci. 40, 99, 1975), and TESNaK. The first ‘two fluids have resulted in good fertility and TESNaK in zero fertility (Crabo, Larson, and Graham, Cryobiology 9, 331, 1972). Sperm thawed in OLEP featured the highest uptake of radiolabelled proteins (874%) of the plasma activity and those in TESNaK the lowest (402%). There was no difference in the percentage of normal acrosome (21-25s) or osmotically reactive cells (21-25s) between the fluids but the sperm motility was lower in TESNaK (8 vs 26% ). Furthermore, the fertility of the eight boars was assessed after insemination of 16-37 sows per boar with semen frozen as described above and thawed in BTS. The fertility varied from 23 to 71% between boars. The correlation between fertility of the individual boars and sperm motility was 0.19, between fertility and percentage normal acrosome ridges 0.29, and between fertility and the “protein uptake” in BTS 0.43. However, a correlation coefficient of 0.77 was obtained between fertility and the protein uptake in TESNaK. Thus, it appears that the less beneficial thawing fluid aided in disclosing differences between boars. It is possible that the composition of the thawing fluids affects the binding mechanisms of surface coating macromolecules in the sperm membrane. Alterations of the sperm surface may result in rapid elimination of the spermatozoa from the female tract. Differences between boars in this respect may account for loss of fertility by certain boars after freezing of the semen. 60. Acrosornal Proteolytic Actioity of FrozenThawed Boar Spermutozoa. K. I. LARS~~N * (Span. B. G. Crabo) (Department of Ob-