02965naa a2200205 a 450000100080000000500110000800800410001902400310006010000160009124500930010726000090020052023690020965000280257865000170260665000100262365000290263365000100266265000170267277300700268915219952023-04-27       1964    bl uuuu     u00u1 u    #d7 a10.1530/jrf.0.00802572DOI1 aDOTT, H. M.  aResults of inseminations with ram spermatozoa in different media.h[electronic resource]  c1964  aAbstract: Previous studies have shown that stored ram semen (ram spermatozoa kept in contact with seminal plasma for 4\m=1/2\hr at 30\s=deg\C) is less likely to cause conception when inseminated into a ewe than fresh semen with a lower impedance change frequency (Dott, 1961). In the same paper it was shown that epididymal semen could be stored at 30\s=deg\C and ejaculated semen at 20\s=deg\C for 4\m=1/2\hr without causing a serious drop in the conception rate obtained with them. It could be concluded from these experiments that some factor in the seminal plasma adversely affected the fertilizing ability of ram spermatozoa. This effect is dependent on time and temperature but it is not known whether it is dependent on the continued presence of seminal plasma. Ewes have been inseminated with ejaculated ram spermatozoa separated from the seminal plasma, as the first in a series ofexperiments to test this. In these preliminary experiments the spermatozoa were freed of seminal plasma by a washing procedure, the first stage ofwhich was a 1 : 1 dilution and the final theoretical dilution of the constituents of the seminal plasma was 1 : 2592. This was brought about in four identical cycles of dilution and centrifugation at about 300 g. In practice the theoretical dilution is not achieved for constituents of the seminal plasma (Dott & Walton, 1960). The sperm density of the final suspension was the same as in the ejaculated semen. The effects offour washing fluids were studied, two phosphate buffers and two bicarbonate buffers. The composition of the phosphate buffers was (i) sodium phosphate: 0\m=.\128m-NaCl, 0\m=.\0204m-Na2HPO4.12H2O, 0\m=.\0048m-NaH2PO4. 2H2O; (ii) sodium potassium phosphate: 0\m=.\124m-Nacl, 0\m=.\005m-KCl, 0\m=.\0204m-Na2HPO4.12H2O, 0\m=.\0048m-NaH2PO4.2H2O and the bicarbonate buffers contained (iii) sodium bicarbonate: 0\m=.\128m-Nacl, 0\m=.\026m-NaHCO3; (iv) sodium potassium bicarbonate; 0\m=.\124m-NaCl, 0\m=.\005m-KCl, 0\m=.\026 m-NaHCO3. All were adjusted to pH 7\m=.\3. After washing in sodium potassium bicarbonate, the spermatozoa were packed into a pellet which was more difficult to disperse than the loose aggregations obtained in the other three solutions. The results of insemination are given in Table 1 and an overall X2 analysis showed that the spermatozoa washed in phosphate buffer produced a higher  aArtificial insemination  aReproduction  aSheep  aInseminação Artificial  aOvino  aReprodução  tJournal Reproduction and Fertilitygv. 8, n. 2, p. 257-258, 1964.