Bovine spermatozoal head size variation and evaluation of a separation technique based on this size.

The objectives of this study were to determine the variation of head areas of normal spermatozoa attributable to breed, individual bull and ejaculate and to verify separation of X and Y chromosome-bearing spermatozoa and separation effectiveness. Spermatozoa were evaluated using video enhanced contrast microscopy combined with video intensified fluorescent microscopy and the polymerase chain reaction (PCR). In Experiment 1, spermatozoal head areas were measured from 2 ejaculates collected from bulls of 3 beef and 2 dairy breeds. No differences in head areas were found between breeds or between bulls within breeds; variation was observed among ejaculates from individual bulls across breeds. In Experiment 2, spermatozoa from 5 ejaculates were separated on individual SEPDEVICEs (Patented). Head area, fluorescent intensity and PCR of spermatozoa retained in the SEPDEVICEs suggested a separation based on size in 1 of 5 samples. Ejaculate variation in head areas affected separation efficiency.

Sex ratio variation between ejaculates within sire evaluated by polymerase chain reaction, calving, and farrowing records.

Ejaculates from sires were examined by polymerase chain reaction to determine percentage of sperm bearing the Y chromosome. Results were verified by examining the percentage of male calves per ejaculate used in artificial insemination (AI) and the percentage of male piglets per litter from a controlled mating program. Spermatozoal DNA was amplified by polymerase chain reaction with specific primers for the Y chromosome. Image analysis measured the fluorescent intensity of the 194-bp band. Ejaculates were compared with a pooled standard of spermatozoal DNA equated to a 50% Y-bearing sperm ejaculate. Calving data were obtained from information collected for the National Association of Animal Breeders for dystocia evaluation of cows bred to AI bulls. Breeding data were obtained from AI technician receipts. Calving and breeding data were merged on cow, sire, calving date, and breeding date. The percentage of males were calculated per sire, ejaculate, and herd combination. Farrowing data were evaluated for the percentage of male piglets per litter. Ejaculates within bulls contributed to variation (24 +/- 9.8% to 84 +/- 9.8%) in the percentage of sperm bearing the Y chromosome. Ejaculates from the same bull contributed to variation in the percentage of male calves (16.1 to 72.3%). Ejaculates from the same boar contributed to variation in the percentage of male piglets that ranged from 7.8 to 94.7%. These percentages and the results obtained by polymerase chain reaction analysis of ejaculates suggested that spermatozoa bearing X and Y chromosomes were unequally represented in ejaculates. The use of ejaculates screened by polymerase chain reaction could enhance production of the desired sex of calf.