New protocol to separate llama sperm without enzymatic treatment using Androcoll‐E ™

BERTUZZI, MARIANA L.; FUMUSO, FERNANDA G.; GIULIANO, SUSANA M.; MIRAGAYA, MARCELO H.; GALLELLI, MARÍA F.; CARRETERO, MARÍA I.

REPRODUCTION IN DOMESTIC ANIMALS (1990)

WILEY-BLACKWELL PUBLISHING, INC

Año: 2020 vol. 55 p. 1154 - 1162

0936-6768

The objective of this study was to design a protocol to separate spermatozoa fromseminal plasma of raw llama semen without prior enzymatic treatment using a singlelayer centrifugation with Androcoll-E? (AE). Two experiments were performed: (a)samples were divided into three aliquots (1 ml) that were deposited on the top of 4, 5or 6 ml of AE and were centrifuged at 800g for 20 min and (b) samples were dividedinto two aliquots (1 ml) that were deposited on the top of 4 ml of AE and were centrifuged at 600g or 1,000g for 20 min. Columns of 5 and 6 ml of AE showed a total sperm motility (TM) significantly lower, while in the 4 ml column, this parameter was not different from the TM of samples before the AE treatment. The percentage of spermatozoa with intact and functional membranes, normal morphology and intact acrosomes, as well as the percentages of sperm with highly condensed chromatin, was conserved (p ˃ .05) in the three column heights and in the two centrifugationspeeds evaluated. In conclusion, the different column heights of AE (4, 5 and 6 ml)and the different centrifugation speeds used (600, 800 and 1,000g) allow separating spermatozoa of raw llama semen without enzymatic treatment, preserving the evaluated sperm characteristics. However, of all the studied treatments, centrifugation inthe 4 ml column of AE at 800g would be the method of choice to process raw llamasemen samples destined for reproductive biotechnologies.