Fluorescent multiple staining and CASA system to assess boar sperm viability and membranes integrity in short and long-term extenders

• Main Authors: F. Cremonesi, A. Meucci, A. Lange-Consiglio
• Format: Article
• Language: English
• Published: Tripoli University 2013-03-01
• Series: Open Veterinary Journal
• Subjects: Boar; CASA; Fluorescent staining; Motility; Semen storage.
• Online Access: http://www.openveterinaryjournal.com/2013/Volume%203%20(1)/OVJ-152-12-12%20A.%20Lange-Consiglio%20et%20al.pdf

The aim of this study was to assess the effect on boar spermatozoa quality of in vitro storage in short and long-term extenders by fluorescent multiple staining (FMS) and computer assisted semen analyzer (CASA). Fresh ejaculates from three healthy, sexually mature boars were diluted with equal volumes of six short-term or three long-term commercial extenders and stored at 19°C for 6 days (short-term) or 12 days (long-term). The integrity of spermatozoa membranes was analyzed by FMS using propidium iodide, 5,5’,6,6’-tetrachloro-1,1’,3,3’ tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) and fluorescein isothiocyanate-conjugated peanut agglutinin (PNA). The results obtained from this staining were compared with spermatozoa motility assessed by CASA. Our study showed that the number of viable spermatozoa with non-reacted acrosomes and intact mitochondria was positively correlated with the rate of motile spermatozoa (r2>0.9) irrespective of the extender used. In all extenders the number of motile spermatozoa significantly decreased as preservation period increased (P<0.05). FMS test is a potent indicator of sperm motility because it analyses mitochondrial integrity independently from observable alterations in motility. The best performing extenders were BTS for short-term storage and TRI-x-Cell for long-term storage.