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Research article2009Peer reviewed

Single-layer centrifugation with Androcoll-E can be scaled up to allow large volumes of stallion ejaculate to be processed easily

Morrell, Jane; Johannisson, Anders; Dalin, Anne-Marie; Rodriguez, Heriberto

Abstract

The objective of the current study was to optimize the volumes of Androcoll-E and sperm sample used in various sizes of centrifuge tube to scale up single-layer centrifugation (SLC) for routine use in the field. Although sperm suspensions of equivalent quality were produced using Androcoll-E in small and large tubes, the sperm yield was much lower in the latter (P < 0.001). In contrast, in 200-mL tubes (XL), the yields were approximately 25% higher than those for the small tubes. An increased volume (4.5 mL) of extended ejaculate in small tubes (SLC-Inc) or 15 to 18 mL extended ejaculate on 15 mL of colloid of a reduced density, Androcoll-E-Large (SLC-Large), in 50-mL tubes were both found to give similar yields of motile spermatozoa as that of the SLC-Small method (SLC-Small, 49.7 +/- 18.6%; SLC-Inc, 53.3 +/- 17.1%; SLC-Large, 44.9 +/- 18.3%) and were found to be equivalent in quality (motility: 88.0 +/- 8.8%, 84.0 +/- 3.5%, 90.0 +/- 5.4%; normal morphology: 69.4 +/- 17.0%, 69.4 +/- 12.7%, 63.9 +/- 15.6%; viability: 78 +/- 16.7%, 83.8 +/- 12.5%, 80.05 +/- 14.6%; DNA fragmentation index: 14.7 +/- 10.9%, 12.8 +/- 8.1%, 11.6 +/- 7.6%, respectively). The processing of an "average" stallion Equus caballus ejaculate in approximately twenty-seven 10-mL tubes (SLC-Inc) or eight 50-mL tubes (SLC-Large) is feasible, the latter being considered more practical for on-stud use. (C) 2009 Elsevier Inc. All fights reserved.

Keywords

Chromatin integrity; Morphology; Scale up; SLC; Sperm motility; Stallion

Published in

Theriogenology
2009, Volume: 72, number: 6, pages: 879-884
Publisher: ELSEVIER SCIENCE INC