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United States Department of Agriculture

Agricultural Research Service

Research Project: National Animal Germplasm Program

Location: Plant And Animal Genetic Resources Preservation Research Unit

Title: Effect of storage time on the viability of cryopreserved bovine spermatozoa

Authors
item Gallegos, A -
item Ericsson, S -
item Blackburn, Harvey
item Spiller, Scott
item Warnock, B -
item Meador, M -
item Smith, M -
item Purdy, Phil

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: July 8, 2013
Publication Date: July 8, 2013
Citation: Gallegos, A.I., Ericsson, S.A., Blackburn, H.D., Spiller, S.F., Warnock, B.J., Meador, M.K., Smith, M.W., Purdy, P.H. 2013. Effect of storage time on the viability of cryopreserved bovine spermatozoa. Meeting Abstract. American Society of Animal Science, Indianapolis, IN, July 8-12, 2013.

Technical Abstract: Long term cryopreserved semen viability can impact the National Animal Germplasm Program’s (NAGP) sampling strategy and ability to reconstitute livestock populations. Therefore, the purpose of this project was to determine if prolonged storage of cryopreserved sperm impacts cell viability. Cryopreserved sperm samples from 12 Hereford bulls were utilized from the NAGP repository. These samples were separated into groups based on storage time of 40-50, 30-39, 20-29 or 10-19 yr. The percentage of progressively (PMS) and total motile sperm (TMS), curvilinear velocity (VCL), and beat cross frequency (BCF) was obtained using computer assisted sperm analysis. Flow cytometric analysis and specific fluorescent stains were utilized for the following viability assessments: FITC PNA –percent live non-acrosome reacted sperm (LNARS); propidium iodide – percent membrane intact sperm (MIS); Yo-Pro-1 - cell membrane integrity/ non-apoptotic sperm (NAS); merocyanine 540 (M540) - percent of sperm with relatively ordered membranes (ROM); and the combination of Yo-Pro-1 with M540 for assessment of membrane phospholipid order (MPO). Differences in measures of sperm viability were assessed using ANOVA with a fixed effect model that included the effects of storage time, type of cryopreservation diluent (milk or egg yolk), and their interaction. Significant differences were not observed for the main effects or for the interaction term (Table 1). These results demonstrate that bovine sperm viability was not impacted as a result of the storage time in liquid nitrogen. These results suggest sample deterioration is not occurring and need not be a factor for consideration in collection development and sample utilization.

Last Modified: 9/10/2014