Assessment of selected quality parameters of epididymal cat (Felis catus s. domestica, L. 1758) sperm using flow cytometry method and computer assisted sperm analyser.
The improvement of biotechnical methods connected with fast and precise semen quality assessment and its utilization in assisted reproductive techniques is an urgent necessity in felids. The aim of this study was to evaluate some quality parameters (i.e. the viability and share of cells with intact plasma membrane) of epididymal sperm of cats using the flow cytometry method and computer-assisted sperm analysis (CASA) examination. The material consisted of epididymal spermatozoa flushed from 22 pairs of epididymes after routine neutering procedures obtained from domestic cats aged between 8 and 36 months. The epididymes were cut and incubated with an extender without egg yolk. The samples were assessed for sperm viability (Live/Dead Sperm Viability Kit), percentage of subtle membrane changes (Apoptosis Detection Kit) and motility using FACScalibur flow cytometer and assisted sperm analyser HTM IVOS version 12.2. The flow cytometry method revealed 71.3% and 84.4% of live sperm using Live/Dead Sperm Viability Kit and Apoptosis Detection Kit respectively. The population of early-apoptotic and late-apoptotic sperm were 0.8% and 1.1% respectively. The CASA examination found 51.5% of motile sperm. However, the motility examination under light microscope revealed 69.5% of motile sperm. The data revealed an indistinctive per cent of apoptotic cells and 18.9% and 15.6% of dead cells using Live/Dead Sperm Viability Kit and Apoptosis Detection Kit, respectively, which indicate that the sperm obtained after flushing the epididymis possess potential properties for further assisted reproduction techniques.
Siemieniuch
M J
MJ
Department of Reproductive Immunology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland. martas@pan.olsztyn.pl
Woclawek-Potocka
I
I
D000818:Animals; D017209:Apoptosis; D002415:Cats / Q000502:physiology*; D004822:Epididymis / Q000166:cytology* / Q000502:physiology; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods / Q000662:veterinary*; D007091:Image Processing, Computer-Assisted; D008297:Male; D012661:Semen / Q000166:cytology / Q000502:physiology; D013076:Sperm Count / Q000295:instrumentation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility / Q000502:physiology; D013094:Spermatozoa / Q000502:physiology*