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

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Author(s): A. Lange-Consiglio | A. Meucci | F. Cremonesi

Journal: Open Veterinary Journal
ISSN 2218-6050

Volume: 3;
Issue: 1;
Start page: 21;
Date: 2013;
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Keywords: Boar | CASA | Fluorescent staining | Motility | Semen storage.

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

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