Open Veterinary Journal

Peer-Reviewed Journal

 

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

A. Lange-Consiglio, A. Meucci and F. Cremonesi*

Università degli Studi di Milano, Faculty of Veterinary Medicine, Department of Health, Animal Science and Food Safety, Large Animal Hospital, Reproduction Unit, 6 Via dell'Università, I-26900 Lodi, Italy

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Abstract

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.

Keywords: Boar, CASA, Fluorescent staining, Motility, Semen storage.

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Cite this paper:

Lange-Consiglio, A., Meucci, A. and Cremonesi, F. 2013. Fluorescent multiple staining and CASA system to assess boar sperm viability and membranes integrity in short and long-term extenders. Open Vet. J. 3(1), 21-35.