Implementation of flow cytometry for quality control in four Danish bull studs.
A flow cytometric method for simultaneous determination of sperm concentration and viability has recently been developed. In 2001, four Danish bull studs purchased flow cytometers and eight technicians were trained for routine analysis of raw and frozen-thawed semen. After initial training of the technicians, an experiment was carried out to document the precision of the system. The aim was also to assess if flow cytometric determination of sperm concentration could result in a more uniform production of semen doses. Results of this experiment showed high precision in the determination of sperm concentration and coefficients of variation were 3.5 and 2.4% for raw and frozen-thawed semen, respectively. Sperm viability was also assessed with high precision and coefficients of variation were 0.9% for raw semen and 1.7% for frozen-thawed semen. Furthermore, the experiment showed that package of semen doses after flow cytometric determination of sperm concentration in the raw semen results in a significantly smaller variation in the number of sperm per dose. In the second experiment, frozen semen was exchanged between the participating studs and were analysed by flow cytometry as well as by microscopic assessments by the eight technicians. Results show that the average correlation between technicians were 0.38 for motility assessments while flow cytometric agreement between technicians was significantly higher (average correlation was 0.86 for sperm viability and 0.92 for sperm concentration). The experiment also showed very high agreements between assessments within lab technician (correlations r=0.98 (sperm concentration) and r=0.99 (sperm viability)). Experiment 3 revealed that straws from the same batch varies in both concentration and viability. It is concluded that flow cytometric determination of sperm concentration and viability can be used to improve semen assessment by AI studs and result in a better quality control.
Christensen
P
P
Department of Large Animal Sciences, Veterinary Reporduction and Obstetrics, The Royal Veterinary and Agricultural University, Dyrlaegevej 68, 1870 Frederiksberg C, Denmark. PC@kvl.dk
Hansen
C
C
Liboriussen
T
T
Lehn-Jensen
H
H
D000818:Animals; D001947:Breeding; D002417:Cattle; D002470:Cell Survival; D003718:Denmark; D005434:Flow Cytometry / Q000379:methods*; D007315:Insemination, Artificial / Q000662:veterinary; D008297:Male; D011786:Quality Control; D012680:Sensitivity and Specificity; D013076:Sperm Count; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology*