The aim of this study was to determine the percentage of epidydimal sperm acrosome integrity using alpaca Arachis hypogaea (PNA) and Pisum sativum (PSA), combined with fluorescein isothiocyanate (FITC). Testicles (n=45) were obtained at Ninacaca municipal slaughterhouse (Pasco, Peru). Only 29 samples with motility higher than 30% and concentration higher than 50x106 sperm/ml were used. Sperm from cauda epididymis were recovered with 1 ml of Tris base solution, and then, washed by centrifugation at 600 g for 8 min and pellets were re-suspended in 300 μl of PBS. Each sample was divided into two aliquots and incubated for 8 min at 38 °C with FITC-PNA (0.5 μg/ml) or FITC-PSA (2.5 μg/ mL) together with propidium iodide (PI, 0.5 μg/ml) as a viability marker. Samples were evaluated by flow cytometry using a laser excitation 488 nm and emission detector channels 505-560 nm fluorescence (cha...

El objetivo del presente trabajo fue determinar el porcentaje de integridad acrosomal en espermatozoides epididimarios de alpaca utilizando Arachis hypogaea (PNA) y Pisum sativum (PSA), conjugadas con isoticionato de fluoresceína (FITC). Se recolectaron 45 testículos de alpaca obtenidos del Camal Municipal de Ninacaca, provincia de Pasco (Perú) y se utilizaron 29 muestras recuperadas de la cola del epidídimo con motilidad mayor de 30% y concentración mayor de 50x106 espermatozoides/ml. Los espermatozoides fueron suspendidos con 1 ml de solución Tris base, se lavaron por centrifugación a 600 g por 8 min y los pellets fueron re-suspendidos en 300 μl de PBS. Cada muestra fue dividida en dos alícuotas e incubadas por 8 min a 38 °C con FITC-PNA (0.5 μg/ml) o FITC-PSA (2.5 μg/ml), junto con un marcador de vitalidad (ioduro de propidio [PI], 0.5 μg/ml). Las muestras fueron evaluada...

The aim of this study was to demonstrate, by flow cytometry, that the percentage of alpaca spermatozoa with high mitochondrial membrane potential (MMP) is significantly reduced after the cryopreservation process. Forty-one alpaca testicles were obtained from a local slaughterhouse in the Ninacaca district, Pasco, Peru. The spermatozoa were recovered from the tail of the epididymis with an extender based on skim milk and only those samples with minimum 30% motility and 50x106 spermatozoa/ml were cryopreserved. The MMP evaluation was carried out before and after the cryopreservation process. Each sample was incubated for 10 minutes at 38 °C with MitoTracker Deep Red (100 nM) to determine the percentage of high MMP by imaging flow cytometry. The effect of cryopreservation on the percentage of spermatozoa with high MMP was evaluated by a paired t-student test and the percentage of motility ...

The aim of this study was to demonstrate, by flow cytometry, that the percentage of alpaca spermatozoa with high mitochondrial membrane potential (MMP) is significantly reduced after the cryopreservation process. Forty-one alpaca testicles were obtained from a local slaughterhouse in the Ninacaca district, Pasco, Peru. The spermatozoa were recovered from the tail of the epididymis with an extender based on skim milk and only those samples with minimum 30% motility and 50x106 spermatozoa/ml were cryopreserved. The MMP evaluation was carried out before and after the cryopreservation process. Each sample was incubated for 10 minutes at 38 °C with MitoTracker Deep Red (100 nM) to determine the percentage of high MMP by imaging flow cytometry. The effect of cryopreservation on the percentage of spermatozoa with high MMP was evaluated by a paired t-student test and the percentage of motility ...